共查询到20条相似文献，搜索用时 31 毫秒
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AIM To explore the effects of sphingosine kinase 1 (SphK1) on the migration and invasion of non-small-cell lung cancer (NSCLC) cells and its mechanism. METHODS Thirty-one tumor specimens, which were surgically resected and routinely histologically confirmed as NSCLC, and matched adjacent lung tissues were selected. Immunohistochemical staining and RT-qPCR were used to detect the expression of SphK1. The pcDNA3.1-SphK1 vector (SphK1 group), empty pcDNA3.1 vector control (NC group), SphK1 siRNA (siSphK1 group) or control siRNA (siNC group) was transfected into human lung adenocarcinoma A549 cells, and the protein levels of SphK1, E-cadherin, fibronectin and p-ERK1/2 were determined by Western blot. The effects of over-expression of SphK1 and inhibition of ERK1/2 on migration and invasion of A549 cells were evaluated by Transwell assays. RESULTS SphK1 was highly expressed in the NSCLC tissues and was associated with tumor stage. SphK1 over-expression significantly promoted the migration and invasion of A549 cells, increased the protein levels of p-ERK1/2 and fibronectin, and decreased the protein expression of E-cadherin (P <0.05), but the opposite result was observed after SphK1 interference. The ERK1/2 inhibitor U0126 significantly inhibited the up-regulation of p-ERK1/2 and fibronectin levels and the down-regulation of E-cadherin expression induced by SphK1 over-expression, and also inhibited the invasion and migration of A549 cells promoted by SphK1 over-expression (P <0.05). CONCLUSION SphK1 may reduce E-cadherin protein levels, increase fibronectin protein levels, and promote the invasion and migration of NSCLC cells through ERK1/2 signaling pathway. 相似文献
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LIU Hua ZHANG Su-lan LIANG Tian-song WANG Juan ZHAO Jing-yi ZHENG Ying-juan ZHANG Xiang-xian YANG Dao-ke 《园艺学报》2000,36(10):1789-1795
AIM To investigate the expression level of long noncoding RNA （lncRNA） TTN antisense RNA 1 （TTN-AS1） in lung adenocarcinoma tissues and the effects of TTN-AS1 silencing on the viability and invasion of lung adenocarcinoma A549 cells. METHODS RT-qPCR was used to detect the expression of TTN-AS1, microRNA-519d-3p （miR-519d-3p） and matrix metalloproteinase 2 （MMP2） mRNA in 32 cases of lung adenocarcinoma and adjacent normal tissues. The untransfected A549 cells were divided into blank group, si-NC group （with si-NC transfection） and si-lncRNA group （with silencing of lncRNA TTN-AS1 expression）, with n =5 in each group. The effects of TTN-AS1 silencing on the viability and invasion of A549 cells were detected by CCK8 and Transwell methods. The targeting regulatory effects of TTN-AS1 on miR-519d-3p and miR-519d-3p on MMP2 were determined by dual-luciferase reporter assay, RNA immunoprecipitation test, RT-qPCR and Western blot. RESULTS The expression level of TTN-AS1 in 32 cases of lung adenocarcinoma tissues is notably higher than that in the adjacent normal tissues （P <0.05）. Silencing of TTN-AS1 in A549 cells significantly suppressed the cell viability and invasion. TTN-AS1 negatively regulated the expression of miR-519d-3p via sponging and absorbing miR-519d-3p. MMP2 is the target gene of miR-519d-3p and can be negatively regulated by miR-519d-3p. Overexpression of MMP2 partially reversed the inhibitory effect of TTN-AS1 silencing and miR-519d-3p overexpression on the invasion of A549 cells. CONCLUSION The lncRNA TTN-AS1 is overexpressed in lung adenocarcinoma tissues, and it regulates lung adenocarcinoma A549 cell viability and invasion via miR-519d-3p/MMP2 pathway. 相似文献
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CONG Wen-wen FENG Ye-nan WANG Shuai-xing HU Guo-min XIAO Han SI Jun-qiang ZHANG You-yi 《园艺学报》2021,37(1):27-33
AIM To investigate the effects of paired box 6 （PAX6） on angiotensin II （Ang II）-induced transdifferentiation of cardiac fibroblasts （CFs） and its underlying mechanisms. METHODS Primary CFs were isolated from the hearts of adult mice, and Ang II was used to induce the transdifferentiation of CFs. The adenovirus vector carrying PAX6 was constructed and transfected into the CFs. The cells were divided into Ad-GFP+Ctrl group （transfected with control adenovirus vector）, Ad-GFP+Ang II group （transfected with control adenovirus vector and treated with Ang II）, Ad-PAX6+Ctrl group （transfected with adenovirus vector carrying PAX6 ） and Ad-PAX6+Ang II group （transfected with adenovirus vector carrying PAX6 and treated with Ang II）. The fluorescence expressed by transfected CFs was observed under an inverted fluorescence microscope. The protein levels of PAX6, α-smooth muscle actin （α-SMA）, collagen type I （Col I）, fibronectin （FN） and transforming growth factor β1 （TGFβ1） were detected by Western blot. The expression and distribution of α-SMA, Col I and FN were measured by immunofluorescence staining. The mRNA levels of PAX6 and TGFβ1 were determined by qPCR. RESULTS The fluorescence observed by inverted fluorescence microscopy confirmed the successful transfection of adenovirus vector into the CFs. qPCR and Western blot showed successful PAX6 overexpression in the CFs （P< 0.01）. Ang II increased the myofibroblast marker α-SMA in the CFs, while overexpression of PAX6 significantly inhibited the expression of α-SMA induced by Ang II （P< 0.01）. In addition, PAX6 overexpression significantly inhibited Ang II-induced synthesis of extracellular matrix （ECM） proteins Col I and FN （P< 0.05）. Furthermore, Ang II treatment upregulated TGFβ1 mRNA and protein levels, while overexpression of PAX6 significantly inhibited TGFβ1 mRNA and protein expression induced by Ang II （P< 0.05）. CONCLUSION PAX6 inhibits Ang II-induced CF transdifferentiation and ECM protein synthesis via inhibiting TGFβ1 expression. 相似文献
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LI Jun ZHANG Qian MA He CHEN Chu-jie YANG Xiang-wei PANG Jun JIANG Dong-gen 《园艺学报》2000,36(9):1572-1578
AIM To investigate the expression of pyruvate dehydrogenase kinase 4 （PDK4） in prostate cancer tissue and its effect on glycolysis and growth of prostate cancer cells. METHODS Immunohistochemistry was used to compare the expression differences of PDK4 protein in benign prostatic hyperplasia （BPH） and prostate cancer tissues. The expression levels of PDK4 in normal prostatic epithelial cells （RWPE-1） and different prostate cancer cell lines （PC3, LNCaP, DU145 and C4-2） were detected by RT-qPCR and Western blot. Recombinant plasmid carrying PDK4-shRNA was constructed, and the expression of PDK4 in prostate cancer PC3 cells was down-regulated by transfection with PDK4-shRNA. The changes in glycolysis level of PC3 cells before and after transfection were determined by cell glycolysis kit, and the effects of PDK4 on the viability and cell cycle distribution of PC3 cells were detected by CCK-8 assay and flow cytometry. RESULTS In prostate cancer tissues, the expression level of PDK4 protein was significantly higher than that in BPH tissues （P <0.05）, and the analysis of immunohistochemical score showed that prostate cancer tissues with high Gleason score displayed significantly higher PDK4 expression than those with low Gleason score （P <0.05）. Compared with normal prostatic epithelial cells, RT-qPCR and Western blot results indicated that the expression level of PDK4 was also significantly increased in prostate cancer cell lines （P <0.05）. In addition, CCK-8 assay results showed that the viability of prostate cancer PC3 cells was significantly decreased after knockdown of PDK4 expression （P <0.05）. The results of flow cytometry demonstrated that knockdown of PDK4 expression in PC3 cells resulted in a notable increase in G0/G1 phase arrest （P <0.05）. CONCLUSION PDK4 is highly expressed in prostate cancer tissues and cell lines, and significantly increases in prostate cancer with high Gleason score. In addition, down-regulation of PDK4 expression significantly inhibits glycolysis and growth of prostate cancer cells, resulting in cell cycle arrest at G0/G1 phase. 相似文献
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AIM To investigate the expression of histone chaperone anti-silencing function 1B （ASF1B） in prostate cancer cells and its effect on cell viability in vitro . METHODS Human prostate cancer PC-3 cells were used, and knockdown of ASF1B was conducted by small interfering RNA （siRNA） transfection into the cells. The cells were divided into control group, siRNA negative control vector （mock） group and siRNA-ASF1B group. The viability of the PC-3 cells treated with ASF1B-siRNA for 12, 24 and 48 h was measured by MTT assay. Flow cytometry was used to analyze cell apoptosis and cell cycle distribution. The mRNA expression of apoptosis-related molecules was detected by RT-qPCR, and the expression levels of MAPK/JNK/ERK signaling pathway-related proteins were determined by Western blot. RESULTS The protein level of ASF1B in the normal cells （benign prostatic hyperplasia） was significantly lower than that in the PC-3 cells （P <0.01）. Compared with control group and mock group, the protein expression level of ASF1B in the PC-3 cells transfected with siRNA-ASF1B plasmid and the viability of the PC-3 cells were significantly decreased （P <0.01）. The apoptotic rate of the PC-3 cells transfected with siRNA-ASF1B plasmid was significantly higher than that in control group （P <0.01）, and the cell cycle was arrested at G1 phase. The mRNA levels of p53, caspase-3, Bax and PARP-1 in the PC-3 cells transfected with siRNA-ASF1B plasmid were up-regulated compared with those in control group and Mack group （P <0.01）. In addition, the protein levels of MAP2K4 and p-JNK in the PC-3 cells in siRNA-ASF1B group were significantly higher than those in mock group （P <0.01）, while the protein level of p-ERK was significantly lower than that in mock group （P <0.01）. CONCLUSION ASF1B silencing induces G1 arrest and promotes apoptosis of PC-3 cells. Activating MAPK/JNK/ERK signaling pathway may be a possible contributor to the anti-prostate cancer effect of siRNA-ASF1B. 相似文献
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AIM To investigate the effects of long non-coding RNA (lncRNA) LINC01503 on the viability, migration and invasion of lung cancer cells and its mechanism. METHODS Human lung carcinoma H1299 cells were divided into si-NC group (transfected with si-NC), si-LINC01503 group (transfected with si-LINC01503), pcDNA group (transfected with pcDNA), pcDNA-LINC01503 group (transfected with pcDNA-LINC01503), miR-NC group (transfected with miR-NC), miR-335-5p group (transfected with miR-335-5p mimics), si-LINC01503+anti-miR-NC group (co-transfected with si-LINC01503 and anti-miR-NC), si-LINC01503+anti-miR-335-5p group (co-transfected with si-LINC01503 and anti-miR-335-5p), miR-NC+WT-LINC01503 group (co-transfected with miR-NC and WT-LINC01503), miR-NC+MUT-LINC01503 group (co-transfected with miR-NC and MUT-LINC01503), miR-335-5p+WT-LINC01503 group (co-transfected with miR-335-5p and WT-LINC01503) and miR-335-5p+MUT-LINC01503 group (co-transfected with miR-335-5p and MUT-LINC01503). The expression of miR-335-5p and LINC01503 was detected by RT-qPCR. Western blot was used to detect protein expression. MTT assay was used to detect cell viability. Transwell assay was used to detect the migration and invasion abilities. Dual-luciferase reporter assay was used to confirm the targeted relationship between LINC01503 and miR-335-5p. RESULTS Compared with normal tissues, the expression of LINC01503 was significantly increased in the lung cancer tissues, and the expression of miR-335-5p was significantly decreased (P <0.05). Compared with stage I/II , the expression level of LINC01503 in the lung cancer tissues of stage III/IV was significantly increased, and the expression of miR-335-5p was significantly decreased (P <0.05). The patients with high expression of LINC01503 had lower short-term survival rates than those with low expression of LINC01503 (P <0.05). Compared with normal human bronchial epithelial cell line BEAS-2B, the expression of miR-335-5p in lung cancer cell lines H1299, A549 and SPC-A-1 were significantly decreased, and the expression of LINC01503 was significantly increased (P <0.05). Over-expression of miR-335-5p and inhibition of LINC01503 expression inhibited the viability, migration and invasion of H1299 cells, and inhibited the protein expression of cyclin D1, matrix metalloproteinase-2 （MMP-2） and MMP-9 (P <0.05). LINC01503 targeted and regulated miR-335-5p expression, and interfering with miR-335-5p expression reversed the inhibitory effect of inhibiting LINC01503 expression on the viability, migration and invasion of H1299 cells. CONCLUSION Inhibition of lncRNA LINC01503 inhibits the viability, migration and invasion of lung cancer cells. The mechanism may be related to the targeted regulation of miR-335-5p. 相似文献
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AIM To investigate the effects of Triptergium wilfordii multiglucoside （TWM） on intestinal flora and immune function in IgA nephropathy （IgAN） rats based on core 1 β1,3-galactosyltransferase （C1GALT1） and its chaperone protein Cosmc （C1GALT1/Cosmc pathway）. METHODS The rat model of IgAN was established, and the animals were randomly divided into model group （IgAN group）, dexamethasone （Dex） group and TWM group. Normal rats served as normal control （NC） group. The levels of serum creatinine （SCr） and blood urea nitrogen （BUN）, 24-hour urinary total protein （24 h UTP） and the number of urinary red blood cells were measured by automatic biochemical analyzer. The levels of serum IgA1, and plasma tumor necrosis factor-α （TNF-α）, B-cell activating factor （Baff） and interleukin-17 （IL-17） were detected by ELISA. The level of galactose-deficient IgA1 （Gd-IgA1） was detected by Vicia villosa lectin affinity ELISA. The intestinal colony was cultured in selective bacterial medium. The ratio of CD4+ CD25+ regulatory T cells （Treg） to CD4+ T cells （Treg proportion） in peripheral blood mononuclear cells （PBMC） was detected by flow cytometry.Western blot was used to determine the protein expression of C1GALT1 and Cosmc in intestinal mucosa. RESULTS Compared with NC group, 24 h UTP, the number of urinary red blood cells, SCr, BUN, serum IgA1 and Gd-IgA1, the numbers of Enterobacteriaceae , Enterococcus and Bacteroides , and the levels of TNF-α, Baff and IL-17 in plasma in IgAN group were significantly increased （P <0.05）, while the numbers of Bifidobacteria and Lactobacilli , the Treg proportion in PBMC, and the protein expression levels of C1GALT1 and Cosmc in intestinal mucosa were significantly decreased （P <0.05）. Compared with IgAN group, 24 h UTP, the number of urinary red blood cells, SCr, BUN, serum IgA1 and Gd-IgA1, the numbers of Enterobacteriaceae , Enterococcus and Bacteroides , and the levels of TNF-α, Baff and IL-17 in plasma in Dex group and TWM group were significantly reduced （P <0.05）, and those in TWM group were lower than those in Dex group （P <0.05）. Moreover, the numbers of Bifidobacteria and Lactobacilli , the Treg proportion in PBMC, and the protein expression levels of C1GALT1 and Cosmc in intestinal mucosa were significantly elevated （P <0.05）, and those in TWM group were higher than those in Dex group （P <0.05）. CONCLUSION TWM reduces the abnormal glycosylation level of IgA in IgAN rats by promoting the activation of C1GALT1/Cosmc pathway, and attenuates the intestinal flora disorder and immune dysfunction in IgAN rats, thus exerting the therapeutic effect. 相似文献
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WANG Xue-ting ZHANG Li-ping ZHOU Dan-dan ZHENG Quan MU Qing-jie ZHANG Bao-gang LI Hong-li YIN Chong-gao 《园艺学报》2000,36(10):1881-1886
AIM To explore the expression of perilipin 3（PLIN3） in lung adenocarcinoma and the relationship between the prognosis of patients and the invasiveness of lung adenocarcinoma cells. METHODS HPA database was used to predict the genes related to poor prognosis of lung cancer and PLIN3 was selected as the research object.HPA database was used to analyze the correlation between PLIN3 and survival rate of lung cancer and lung adenocarcinoma.GEPIA database was used to further verify the correlation between the expression difference of PLIN3 and the survival rate of lung adenocarcinoma patients. The expression of PLIN3 in lung adenocarcinoma was further analyzed by using Ualcan database.Western blot was used to detect the expression of PLIN3 in lung adenocarcinoma cells.siPLIN3 plasmid was constructured and Transwell assay was used to detect the invasion ability of A549 cells after transfection. RESULTS PLIN3 was significantly related to the survival rate of the patients with lung adenocarcinoma and it was over-expressed in lung adenocarcinoma tissues.The expression of PLIN3 was closely related to the stages of cancer and the grades of lymph node metastasis of lung adenocarcinoma.PLIN3 was over-expressed in lung adenocarcinoma cells. The number of the A549 cells passing through Transwell in knock-down group was significantly lower than that in control group. CONCLUSION PLIN3 is highly expressed in lung adenocarcinoma. The expression level of PLIN3 is related to the prognosis of lung adenocarcinoma patients.Knock-down of PLIN3 inhibits the invasion ability of lung adenocarcinoma cells in vitro . 相似文献
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MA Dong-xue GAO Wei-juan LIU Zhen-yi ZHANG Hong-jun WANG Jia-ying DONG Xian-hui 《园艺学报》2021,36(12):2198-2204
AIM To explore the effect of compound of Epimedium , Astragalus and Radix Puerariae on the expression of a disintegrin and metalloproteinase 10 （ADAM10） in Aβ-induced hippocampal neuron HT22 cells with or without hepcidin （HAMP） expression knock-down for analyzing the pathogenesis of Alzheimer disease （AD） at cell level. METHODS Hippocampal neuron HT22 cells were cultured in vitro and randomly divided into 7 groups： control group, Aβ group （Aβ25-35-induced HT22 cells）, RNAi group （HAMP gene was silenced in HT22 cells）, Aβ+RNAi group （HAMP gene expression in Aβ25-35-induced HT22 cells was silenced）, Aβ+TCM group （Aβ25-35-induced HT22 cells were treated with Epimedium , Astragalus root and Radix Puerariae effective components）, RNAi+TCM group （HT22 cells with HAMP gene silence were treated with Epimedium , Astragalus root and Radix Puerariae effective components） and Aβ+RNAi+TCM group （Aβ25-35-induced HT22 cells with HAMP gene silence were treated with Epimedium , Astragalus root and Radix Puerariae effective components）. The silence efficiency of HAMP siRNA was detected by qPCR and Western blot. The ADAM10 expression in each group was determined by immunofluorescence, qPCR and Western blot. RESULTS The HAMP siRNA-3 sequence had the highest interference efficiency. Compared with control group, the expression levels of ADAM10 in Aβ group, RNAi group and Aβ+RNAi group were decreased （P <0.05）. Compared with Aβ group,the expression levels of ADAM10 in Aβ+RNAi group was also decreased （P <0.05）, and the expression levels of ADAM10 in Aβ+TCM group was increased （P <0.05）. Compared with RNAi group, the expression levels of ADAM10 in Aβ+RNAi group was decreased （P <0.05）, while the expression levels of ADAM10 in RNAi+TCM group was increased （P <0.05）. Compared with Aβ+RNAi group, the expression levels of ADAM10 in Aβ+RNAi+TCM group was increased （P <0.05）. CONCLUSION The effective components of Epimedium , Astragalus and Radix Puerariae compound promotes the expression of ADAM10 in Aβ25-35-induced HT22 cells, which mechanism may be related to the expression of HAMP. 相似文献
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AIM To investigate the expression of baculoviral inhibitor of apoptosis protein repeat-containing protein 5 （BIRC5） in gastric cancer tissue and its relationship with prognosis of gastric cancer patients, and to explore the effect of BIRC5 knock-down on the viability and apoptosis of gastric cancer cells. METHODS The expression of BIRC5 was detected by immunohistochemistry in 67 cases of gastric cancer tissues and paracancerous tissues for analyzing the relationships with clinicopathological characteristics. The mRNA and protein expression levels of BIRC5 in gastric carcinoma cell lines （AGS, MKN-1 and MGC-803） and normal gastric epithelial cell line GES-1 were detected by RT-qPCR and Western blot. The AGS cells were divided into blank group （no treatment）, Ctr-sh group （blank plasmid transfection） and BIRC5-sh group （BIRC5-shRNA plasmid transfection）. The interference efficiency of BIRC5-shRNA was evaluated by Western blot. The cell viability was measured by MTT assay, the apoptosis was analyzed by flow cytometry, and the levels of apoptosis-related proteins cleaved caspase-3, Bax and Bcl-2 were determined by Western blot. RESULTS BIRC5 was mainly expressed in cytoplasm, and the positive expression rate of BIRC5 in the gastric cancer tissues was higher than that in the adjacent tissues （P <0.01）. The positive rates of BIRC5 in the gastric cancer patients at TNM Ⅲ~Ⅳ stages and with lymph node metastasis were higher than those in the patients at TNM Ⅰ~Ⅱ stages and without lymph node metastasis, respectively （P <0.05）. The survival time of the patients with positive BIRC5 expression was shorter than that of the patients with negative BIRC5 expression （P =0.011 2）. The cell viability in BIRC5-sh group was lower than that in blank group and Ctr-sh group at time points of 48, 72 and 96 h. The apoptotic rate in BIRC5-sh group was increased compared with blank group and Ctr-sh group. The protein levels of cleaved caspase-3 and Bax in BIRC5-sh group were higher than those in blank group and Ctr-sh group, while the protein expression of Bcl-2 in BIRC5-sh group was lower than that in blank group and Ctr-sh group （P <0.05）. CONCLUSION High expression of BIRC5 in gastric cancer indicates poor prognosis. BIRC5 promotes the growth of gastric cancer cells and inhibits apoptosis. 相似文献
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AIM To construct the mouse embryonic stem cell （ESC） line with stable pancreatic and duodenal homeobox 1 （Pdx1 ） expression by Tet-On system, which may lay a foundation for further research on the differentiation of Pdx1+ definitive endoderm cells into pancreatic cells. METHODS The Pdx1 -overexpressing lentiviral vector with green fluorescent protein marker and puromycin resistance was constructed by Tet-On system and was used to infect the mouse ESC. The cells were divided into 3 groups： blank control group （ESC group）, empty lentivirus control group （PDX1- ESC group） and Pdx1 lentivirus transfection group （PDX1+ ESC group）. Flow cytometry was used to detect the transfected cells after screening by doxycycline （DOX）. The function of Tet-On system and the expression of Pdx1 gene were detected. The transfected cells in PDX1- ESC group and PDX1+ ESC group were sorted by flow cytometry, and constructed ESC line with stable expression of Pdx1 and negative control ESC line were verified. RESULTS （1） The positive rates of transfected cells in PDX1- ESC group and PDX1+ ESC group were 90.72% and 94.01% after screening by DOX, respectively. The positive rates of transfected cells in PDX1- ESC group and PDX1+ ESC group was 97.84% and 98.13% after sorting by flow cytometry, respectively. （2） With DOX, green fluorescence was observed in PDX1- ESC group and PDX1+ ESC group. The mRNA and protein expression of Pdx1 was significantly increased in PDX1+ ESC group （P <0.05）. Without DOX, no green fluorescence was observed in the cells of the 3 groups, and no significant difference in the mRNA and protein expression of Pdx1 was observed （P >0.05）. （3） After 3 months of cryopreservation, the cell lines still survived in resuscitation culture and were regulated by DOX. CONCLUSION Using Tet-On system, the mouse ESC line with inducible Pdx1 expression were successfully established and could be used as an effective cell model to research the differentiation of Pdx1+ definitive endoderm cells into pancreatic cells. 相似文献
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AIM To investigate the effect of stanniocalcin-1 （STC-1） on the proliferation and apoptosis of gastric cancer AGS cells and the role of Bcl-2 in these processes. METHODS The AGS cells were transfected with the plasmids for STC -1 knockdown or over-expression. The cell proliferation was measured by MTT assay and colony formation assay. The migration ability was detected by scratch assay. Apoptosis was analyzed by Hoechst 33342 staining and flow cytometry with annexin V-FITC/PI double staining. The protein expression of Bcl-2, survivin, caspase-3 and cleared caspase-3 was determined by Western blot. The mRNA expression levels of STC-1 and Bcl-2 in 20 cases of clinical gastric cancer tissues and adjacent tissues were detected by RT-qPCR, and the correlation between them was analyzed by Pearson method. RESULTS After over-expression of STC-1, the proliferation and migration abilities of the AGS cells were increased, the expression of Bcl-2 and survivin was increased, while the expression of caspase-3 and cleared caspase-3 was decreased （P <0.05）. After knockdown of STC -1 , the proliferation and migration abilities of the AGS cells were decreased, the expression of Bcl-2 and survivin was decreased, while the expression of caspase-3 and cleared caspase-3 was increased （P <0.05）. The mRNA expression levels of STC-1 and Bcl-2 in the gastric cancer tissues were higher than those in the adjacent tissues. Pearson correlation analysis showed that there was a positive correlation between STC-1 and Bcl-2 mRNA expression in the cancer tissues （r =0.308, P =0.011）. CONCLUSION STC-1 may regulate the biological function of gastric cancer cells by altering the expression level of Bcl-2. 相似文献
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GUO Wan-rong CHEN Zong-lan CAO Huan-yi DENG Hong-rong CAI Meng-yin LIANG Hua XU Fen 《园艺学报》2000,36(11):1921-1927
AIM To investigate the alleviating effect of exenatide （Exe）, a glucagon-like peptide-1 （GLP-1） receptor agonist, on the ectopic lipid accumulation in skeletal muscle of ob /ob mice and its mechanism. METHODS Eight-week-old male ob /ob mice and their wild-type （WT） littermates were randomly divided into 3 groups, ob /ob group, ob /ob +Exe group and WT group, and treated with Exe at 24 nmol/kg or the same volume of saline intraperitoneally once daily for 4 weeks. The body weight, fasting blood glucose （FBG） and fat content were measured after the 4-week treatment. The oil red O staining and the quantification of triglyceride （TG） were performed on the skeletal muscle. The serum levels of TG, total cholesterol and free fatty acid （FFA） were also measured by ELISA. The expression levels of AMP-activated protein kinase （AMPK） and lipid metabolism-related proteins were determined by Western blot. Mouse myoblast C2C12 cells were used as an in vitro model to further investigate the effects of Exe. RESULTS As compared with the ob /ob mice treated with saline, 4-week Exe treatment did not reduce body weight, FBG, food intake and fat content in ob /ob mice （P >0.05）. However, serum FFA was decreased （P <0.05）. Oil red O staining and the quantification of TG showed that 4-week Exe treatment significantly attenuated the ectopic lipid accumulation in the skeletal muscle of ob /ob mice （P <0.05）. The results of Western blot showed that the levels of phosphorylated AMPK （p-AMPK） and lipolysis-related proteins were up-regulated, while the lipid synthesis-related proteins were down-regulated by Exe （P <0.05）. Treatment with Exe alleviated the lipid accumulation in the C2C12 cells induced by sodium palmate （P <0.05）, and the effects of Exe on the levels of p-AMPK and lipid metabolism-related proteins in the C2C12 cells were consistent with those in the ob /ob mice （P <0.05）. Treatment with Exe also up-regulated the protein expression of glucose transporter 4 and improved the ability of glucose uptake in the C2C12 cells （P <0.05）. CONCLUSION Short-term Exe treatment attenuates the ectopic lipid accumulation in skeletal muscle of ob /ob mice by up-regulating lipolysis-related proteins and down-regulating lipid synthesis-related proteins, which is independent on body weight loss. 相似文献
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LUO Ming-hao HU Shu-peng WANG Rui-yu Li Chang Lü Ding-yi YANG Xi-yang LUO Su-xin 《园艺学报》2000,36(10):1739-1744
AIM To investigate whether interleukin-1β （IL-1β） regulates endothelial nitric oxide synthase （eNOS） phosphorylation at Ser1177 site in human umbilical vein endothelial cells （HUVECs）, and to explore its possible mechanism. METHODS The HUVECs were randomly divided into normal control group, tumor necrosis factor-α （TNF-α） group, IL-1β group, IL-6 group, SC79 ［protein kinase B （PKB/AKT） specific agonist］ group and SC79+IL-1β group. Western blot was used to determine the protein levels of eNOS, p-eNOS-Ser1177, AKT and p-AKT-Ser473 in the HUVECs. Chemical colorimetry was used to detect the nitric oxide （NO） content in the culture medium of HUVECs. RESULTS No statistically significant difference of p-eNOS-Ser1177 level in HUVECs treated with TNF-α and IL-6 was observed as compared with normal control group （P >0.05）, while the protein level of p-eNOS-Ser1177 in the HUVECs and the content of NO in the culture medium of HUVECs decreased significantly in IL-1β group （P <0.05）, and the protein level of p-AKT-Ser473 in the HUVECs was decreased as compared with normal control group （P <0.05）. The AKT agonist SC79 blocked the down-regulation effect of IL-1β on p-eNOS-Ser1177 level in the HUVECs and NO content in the culture medium of HUVECs （P< 0.05）. CONCLUSION IL-1β down-regulates the protein level of p-eNOS-Ser1177 in HUVECs and affects the activity of eNOS, which may be involved in AKT/eNOS signaling pathway. 相似文献
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AIM To explore the effect of dasatinib on the viability, apoptosis and migration of human renal carcinoma cell lines 786-O and 769-P, as well as the molecular mechanism in vitro . METHODS 786-O cells and 769-P cells were treated with different concentrations （0~2 μmol/L） of dasatinib, and 0 μmol/L dasatinib was used as blank control group. MTT method was used to detect cell viability. Wound healing assay was used to detect the effect of dasatinib on migration. Hoechst 33258 staining was used to observe the effect of dasatinib on apoptosis. Flow cytometry was used to detect the effect of dasatinib on cell cycle. Western blot method was used to detected cell cycle- and apoptosis-related protein levels. RESULTS Dasatinib inhibited viability and migration of 786-O and 769-P cells, and the inhibitory effect of dasatinib increased with the concentration of dasatinib （P <0.05）. The IC50 values of dasatinib against 786-O and 769-P cell lines were （0.958 7±0.028 8） μmol/L and （0.784 3±0.066 0） μmol/L, respectively. After treatment with dasatinib for 24 h, the expression of pro-apoptotic proteins cleaved caspase-3 and cleaved caspase-9 increased significantly （P <0.01）, while the expression of cyclin D1 decreased （P <0.05）. The cycle-related pathway proteins p53 and p21 increased （P <0.05）, while the level of p-AKT was decreased （P <0.05）. CONCLUSION Dasatinib impaired the viability and migration ability of human renal carcinoma cell lines 786-O and 769-P by up-regulating p53 expression and down-regulating AKT phosphorylation. 相似文献
18.
miR-556-3p regulates viability,migration and invasion of endometrial cancer cells by targeting SASH1
AIM To investigate whether microRNA-556-3p （miR-556-3p） regulates the viability, migration and invasion of endometrial cancer cells by targeting SASH1 gene. METHODS The expression of miR-556-3p, and the mRNA and protein levels of SASH1 in endometrial cancer tissues were detected by RT-qPCR and Western blot. Anti-miR-556-3p or pcDNA-SASH1 was transfected into endometrial cancer Ishikawa cells. The cell viability was detected by MTT assay, the migration and invasion abilities of the cells were detected by Transwell chamber method, and the protein expression levels of cyclin D1, p21, matrix metalloproteinase-2 （MMP-2） and MMP-9 were detected by Western blot. StarBase prediction and dual-luciferase reporter experiments were used to analyze the targeting relationship between miR-556-3p and SASH1. Anti-miR-556-3p and si-SASH1 were co-transfected into the Ishikawa cells, and their effects on cell viability, migration and invasion were examined by the methods described above. RESULTS Compared with adjacent tissues, the expression of miR-556-3p in endometrial cancer tissues was increased significantly, and the expression of SASH1 at mRNA and protein levels was decreased significantly （P <0.05）. Inhibition of miR-556-3p expression or induction of SASH1 over-expression obviously reduced the viability of Ishikawa cells, the number of migratory cells, the number of invasive cells and the protein levels of cyclin D1, MMP-2 and MMP-9, and dramatically increased the protein level of p21 （P <0.05）. miR-556-3p targeted SASH1 and negatively regulated its expression. Knock-down of SASH1 expression reversed the inhibitory effect of miR-556-3p expression inhibition on the viability, migration and invasion of Ishikawa cells. CONCLUSION Inhibition of miR-556-3p expression suppresses the viability, migration and invasion of endometrial cancer cells. The mechanism is related to the regulation of its target gene SASH1 . 相似文献
19.
SU Miao-shang XUE Si-chen XU Li REN Xi-kai HUANG Nan TANG Zhu-qin XU Man-huan 《园艺学报》2021,36(12):2212-2219
AIM To investigate the effect of intermittent hypoxia （IH） on bladder detrusor cells apoptosis and calcium channel, and to discuss the regulatory mechanism of Alpiniae oxyphyllae Fructus （AOF）. METHODS IH model of bladder detrusor cells was established by treating the cells with 6 cycles of 5% O2 for 60 min and 20% O2 for 30 min. Human bladder detrusor cells were cultured in vitro , randomly divided into 6 groups, each group had 8 holes. P2X3 receptor antagonist + IH （A） group, M3 receptor antagonist + IH （B） group, β3 receptor antagonist + IH （C） group, AOF + IH （D） group, saline + IH control （NC） group and air simulation control （AC） group were set up. The cells density and morphology were identified by the methods of counting chamber and immunofluorescence light microscopy （LM） after interventions. The apoptosis was analyzed by flow cytometry. Calcium channel expression was detected by patch clamp. RESULTS （1） Compared with the cells in AC group, the cells density and activity were significantly increased in NC group （P <0.05）; some cells appeared protrusions, turned round and blur in cell borders. （2） The results of immunofluorescence for detecting α-SMA protein expression showed that, compared with the cells in group AC, the mean absorbance （MA ） in group NC was significantly increased （F =3.25, P <0.05）; compared with the cells in group NC, that in group A and group D was both decreased significantly （P <0.05）. （3） Compared with the cells in group AC, the apoptotic rate was significantly decreased in group NC （P <0.05）; Compared with the cells in group NC, the apoptotic rates in group A and group D were both significantly increased （P >0.05）. （4） Compared with the cells in group AC, calcium ion channel expression was significantly decreased （P <0.05）. Compared with the cells in group NC, calcium ion channel expression in AOF （100 mg/L） and AOF （50 mg/L） group was significantly increased （P <0.05）. CONCLUSION IH regulates bladder detrusor cells proliferation and apoptosis through P2X3 bladder nerve receptors, high or moderate dose of AOF may change calcium channel and play a protective role in IH induced cell damage. 相似文献
20.
ZENG Jing-jing XU Xiao-xiao SHI Han-qing XU Guang-yu JIN Qi-ke RUAN Yong-xue REN Fang-fang XIE Zuo-yi LI Lei 《园艺学报》2021,36(12):2113-2122
AIM To investigate the effect of cyanidin （Cyn） on pressure overload-induced cardiac remodeling and the underlying mechanism. METHODS Six-week-old male C57BL/6 mice （n =120） were divided into 4 groups： sham group （n =20）, sham+Cyn group （n =20）, transverse aortic constriction （TAC） group （n =40） and TAC+Cyn group （n =40）. The model of cardiac chronic pressure overload was induced by TAC, and the first day of TAC was defined as day 0. The animals in sham+Cyn group and TAC+Cyn group were treated with Cyn dissolved in DMSO and normal saline （5 mg·kg-1·d-1） for 5 d before TAC, while the animals in sham group and TAC group were treated with the same amount of DMSO and normal saline. Four weeks after TAC, the survival rate of the animals in each group was analyzed, the heart function of the mice was measured by ultrasound echocardiography, and the heart weight/body weight and lung weight/body weight were calculated. The cross-sectional area of the cardiomyocytes was measured by wheat germ agglutinin staining and hematoxylin-eosin staining. The degree of cardiac oxidative stress was evaluated by dihydroethidium staining and measurement of superoxide dismutase （SOD） and malondialdehyde （MDA） levels. The cardiomyocyte apoptosis was detected by TUNEL method. The mRNA expression levels of atrial natriuretic peptide （ANP）, brain natriuretic peptide （BNP） and β-myosin heavy chain （β-MHC） were detected by RT-qPCR, and the protein expression levels of Bax, Bcl-2, optic atrophy protein 1 （OPA1） and dynamin-related protein 1 （Drp1） were determined by Western blot. The mitochondrial morphological changes were observed by transmission electron microscopy. RESULTS Compared with TAC group, the survival rate of the mice in TAC+Cyn group was significantly increased （P <0.05）, the myocardial apoptosis, the cross-sectional area of myocardial cells, the heart weight/body weight, the lung weight/body weight, the level of reactive oxygen species and the MDA content were decreased （P <0.05）, and the SOD was activated （P <0.05）. M-mode ultrasound tests showed that Cyn treatment significantly increased left ventricular ejection fraction and left ventricular fractional shortening in the mice after TAC （P <0.05）, while left ventricular end-diastolic diameter and left ventricular posterior wall thickness in diastole were reduced （P <0.05）. Transmission electron microscopic observation showed that the number of myocardial mitochondria was increased and the mitochondrial area was decreased after TAC （P <0.05）, while treatment with Cyn increased the area of myocardial mitochondria and decreased the mitochondrial number （P <0.05）. Compared with sham group, the protein level of OPA1 in TAC group was significantly reduced （P <0.05）, while treatment with Cyn significantly increased the protein level of OPA1. CONCLUSION Cyanidin significantly increases the survival rate, improves the cardiac function and attenuates the cardiac remodeling of the mice after TAC. The mechanism may be related to the inhibition of myocardial mitochondrial OPA1 cleavage and the promotion of mitochondrial fusion. 相似文献