共查询到20条相似文献，搜索用时 31 毫秒
1.
AIM:To investigate the effects of microRNA (miRNA)-483-3p on the growth and migration of human glioma cell line A172 and its potential mechanisms.METHODS:The abundance of miRNA-483-3p in human embryonic kidney 293 cells and different human glioma cell lines (A172,U251 and SHG44) was measured by RT-qPCR.After down-regulation of miRNA-483-3p by transfection of inhibitor in the A172 cells,the cell viability,cell cycle distribution and cell migration were detected by CCK-8 assay,flow cytometry and Transwell assay,respectively.Furthermore,the protein levels of cell cycle-related molecules and epithelial-mesenchymal transition markers were measured by Western blot.Luciferase reporter assay was used to predict and verify the target gene of miRNA-483-3p.RESULTS:miRNA-483-3p was highly expressed in human glioma cells.Knockdown of miRNA-483-3p inhibited A172 cell viability,arrested cell cycle and decreased cell migration rate.Furthermore,the protein levels of cyclin D1,cyclin-dependent kinase 4,phoshorylated retinoblastoma protein,N-cadherin and vimentin were significantly decreased after knockdown of miRNA-483-3p,accompanied with the up-regulation of E-cadherin and β-catenin protein expression.Luciferase reporter assay demonstrated that Smad4 was a potential target gene of miRNA-483-3p.Down-regulation of Smad4 in the A172 cells transfected with miRNA-483-3p inhibitor partially reversed the effect of miRNA-483-3p on cell viability and migration.CONCLUSION:Knockdown of miRNA-483-3p restrains the growth and migration of A172 cells by targeting Smad4. 相似文献
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AIM: To investigate the effects and mechanisms of microRNA-25(miRNA-25) on the proliferation of human esophageal squamous-cell carcinoma cell line TE1. METHODS: The abundance of miRNA-25 in different tissues was measured by RT-PCR. After silencing or over-expression of miRNA-25 with mimics or inhibitor in TE1 cells, the cell proliferation, cell cycle distribution and the expression of cyclin E1 and cyclin-dependent kinase 2(CDK2) at mRNA and protein levels were measured by CCK-8 assay, BrdU detection, flow cytometry, RT-PCR and Western blot, respectively. RESULTS: miRNA-25 was prominent in esophageal mucosal tissue and highly expressed in TE1 cells (P<0.05). Over-expression of miRNA-25 increased TE1 cell proliferation, promoted the cell cycle progression and enhanced the entrance of the cells into S phase (P<0.05). Inverse results were obtained after down-regulation of miRNA-25(P<0.05). Furthermore, the expression of cyclin E1 and CDK2 at mRNA and protein levels was significantly increased after over-expression of miRNA-25, but decreased after down-regulation of miRNA-25(P<0.05). CONCLUSION: miRNA-25 enhances cell cycle transition by increasing the expression of cyclin E1 and CDK2, thus accelerating TE1 cell proliferation. This study provides a novel mechanism by which miRNA-25 increases the proliferation of human esophageal squamous-cell carcinoma cell line TE1, suggesting that down-regulation of miRNA-25 may be a potential new therapeutic strategy for treating esophageal squamous-cell carcinoma. 相似文献
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AIM: To investigate the regulatory effects of microRNA (miR)-130a on the biological characteristics of rat basilar arterial smooth muscle cells (BASMCs) and its underlying mechanisms. METHODS: The expression of miR-130a in rat BASMCs was measured by real-time PCR. After knockdown of miR-130a with inhibitor in the BASMCs, the cell viability, cell cycle distribution and apoptosis were detected by CCK-8 assay and flow cytometry. The expression of cell cycle-and apoptosis-related molecules, such as cyclin D1, cyclin-dependent kinase 2 (CDK2), p21, Bcl-2 and cleaved caspase-3/caspase-3 at protein levels was determined by Western blot. The growth arrest-specific homeobox protein (Gax) expression at mRNA and protein levels was determined by real-time PCR and Western blot. RESULTS: AngiotensionⅡ (AngⅡ) up-regulated the expression of miR-130a and down-regulated the expression of Gax (P<0.05). Transfection with miR-130a inhibitor partly reversed the increase in AngⅡ-induced cell viability and promoted the Gax expression. Furthermore, the early cell apoptotic rate was significantly increased after down-regulation of miR-130a (P<0.05), and the protein levels of cyclin D1, CDK2, Bcl-2 and p-Rb were significantly decreased, accompanied with the up-regulation of p21 and cleaved caspase-3 (P<0.05). CONCLUSION: Down-regulation of miR-130a restrains the viability and promotes the apoptosis of BASMCs by promoting Gax expression and regulating cell cycle-and apoptosis-related molecules, suggesting that miR-130a may be a potential therapeutic target of brain vascular remodeling during hypertension. 相似文献
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YAN Liang CAO Long ZHANG Ming-kang LU Xin-yu LI Yu-meng MA Jin-zhu XU Lei FAN Qian 《园艺学报》2019,35(11):1994-1998
AIM: To investigate the effect of di-indolyl thiozoline (DIIT) on the proliferation of human lung cancer A549 cells. METHODS: The effects of DIIT on the proliferation of human lung cancer A549 cell line were determined by CCK-8 assay and EdU assay. The effects of DIIT on the expression of cyclin-dependent kinase 4 (CDK4), cyclin D1, and the phosphorylation of Akt and mTOR were determined by Western blot. RESULTS: After the A549 cells were treated with DIIT at 12.5, 25, 50 and 100 mg/L, the cell viability detected by CCK-8 assay was decreased by 12%, 27% (P<0.01), 33% (P<0.01) and 52% (P<0.01), respectively, compared with DMSO control group. The EdU positive cell number determined by EdU assay was decreased by 10%, 21% (P<0.05), 26% (P<0.05) and 34% (P<0.01), respectively, compared with DMSO control group. Compared with DMSO control group, DIIT inhibited the phosphorylation of Akt and mTOR and the expression of cyclin CDK4 and cyclin D1 (P<0.05). CONCLUSION: Di-indolyl thiozoline inhibits the proliferation of A549 cells, which may be related to the decreases in phosphorylation levels of Akt and mTOR and the inhibition of cell cycle-related protein expression. 相似文献
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AIM: To investigate the clinical significance of microRNA-326 (miRNA-326) expression in gastric carcinoma and the effect of up-regulation of its expression on the viability and apoptosis of gastric cancer cells. METHODS: The expression of miRNA-326 in 55 tissue samples of gastric cancer was detected by RT-qPCR, and the relationship between the expression and the clinicopathological features was analyzed. The expression of miRNA-326 in gastric cancer BGC-823 cells was detected by RT-qPCR. The BGC-823 cells were transfected by liposome method, and randomly divided into normal control group (untransfected), mimic-NC group (transfected with negative control mimic) and miRNA-326 mimic group (transfected with miRNA-326 mimic). After up-regulation of miRNA-326 expression, the cell viability was measured by CCK-8 assay, and the apoptosis of the cells was analyzed by flow cytometry. The protein levels of matrix metalloprotein 9 (MMP-9), p21, cyclin D1, Bcl-2 and cleaved caspase-3 were determined by Western blot, and the mRNA expression of cyclin D1 was detected by RT-qPCR. Whether CCND1 (the gene of cyclin D1) was the target gene of miRNA-326 was evaluated by dual-luciferase reporter assay. RESULTS: The expression of miRNA-326 in the gastric cancer tissues was significantly lower than that in the adjacent tissues (P<0.05). The miRNA-326 expression had a significant correlation with the tumor size, lymph node metastasis, differentiation, and clinical stages (P<0.05), but it had no correlation with the age and sex of the patients. Moreover, the expression of miRNA-326 was also closely related to the survival rate of the patients (P<0.05). The expression of miRNA-326 in the BGC-823 cells was significantly lower than that in the normal gastric mucosa GES-1 cells (P<0.05). Compared with normal control group, the expression of miRNA-326 in mimic-NC group did not change significantly, while that in miRNA-326 mimic group was increased significantly (P<0.05). Compared with normal control group, the cell viability in miRNA-326 mimic group was significantly decreased, and the apoptosis was increased (P<0.05). In addition, compared with normal control group, the protein levels of MMP-9, cyclin D1 and Bcl-2, and the mRNA expression of cyclin D1 in miRNA-326 mimic group were decreased, while the protein levels of p21 and cleaved caspase-3 were increased (P<0.05). However, no significant difference of above protein and mRNA levels between mimic-NC group and normal control group was observed. Compared with mimic-NC+miR-326 mimic group, the activity of luciferase in the cells transfected with pmiR-CCND1-WT plasmid was significantly decreased (P<0.05), but that in the cells transfected with pmiR-CCND1-Mut plasmid did not change significantly. CONCLUSION: The expression level of miRNA-326 in gastric cancer tissues is low, and it may promote cell viability and inhibit cell apoptosis by targeting CCND1. 相似文献
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WANG Si-si LIAO Xiao-min SHAO Zhong-ming YUAN Jian-ling FENG Mu-yin HA Yan-ping LI Ru-jia SHEN Zhi-hua JIE Wei 《园艺学报》2018,34(12):2172-2179
AIM:To establish SETD2 gene knockout nasopharyngeal carcinoma (NPC) cell strains based on CRIPSR/Cas9 technique and to analyze their proliferation characteristics. METHODS:Sub-quantitative RT-PCR and Western blot were used to detect the expression of SETD2 in immortalized nasopharyngeal epithelial cell line NP-69, well differentiated NPC cell line CNE1, poorly-differentiated NPC cell line CNE2Z and undifferentiated NPC cell line C666-1, and the SETD2 high expression cell line CNE1 was screened. The proliferation ability of CNE1 cells before and after the SETD2 gene knockout was analyzed by CCK-8 and colony formation assay. The cell cycle distribution was detected by flow cytometry, and the expression of cell cycle-related proteins was detected by Western blot. RESULTS:Compared with NP-69 cells, the expression of SETD2 was decreased gradually in CNE1, CNE2Z and C666-1 cells (P<0.01). Based on the CRISPR/Cas9 technique, 2 monoclonal cell strains with SETD2 gene stable knockout, named CNE1-SETD2-KO-#5 and #9, were successfully screened from total 15 monoclones. The results of CCK-8 and plate colony formation assay confirmed that the proliferation ability of CNE1-SETD2-KO-#5 and #9 cells was significantly enhanced compared with CNE1-WT cells (P<0.05). The results of flow cytometry analysis showed that the G1 phase of CNE1-SETD2-KO-#5 and #9 cells was decreased, while the G2/M and S phases were increased significantly (P<0.05). The results of Western blot confirmed the increases in the protein levels of proliferating cell nuclear antigen (PCNA), cyclin D1, cyclin B1, cyclin A2, cyclin E1, cyclin-dependent kinases 2 (CDK2) and CDK4, and the decrease in the protein level of p21 after SETD2 gene knockout (P<0.05). CONCLUSION:The NPC cell strains with SETD2 gene knockout were successfully constructed based on CRISPR/Cas9 technique. SETD2 expression correlates with cell differentiation status in the NPC cells. SETD2 gene knockout promotes NPC cell proliferation by up-regulating cyclin D1, cyclin B1, cyclin A2, cyclin E1, CDK2 and CDK4, and down-regulating p21 expression. 相似文献
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AIM To extract and purify recombinant human progranulin (rhPGRN) and to examine its effect on the proliferation, autophagy and endoplasmic reticulum stress (ERS) in human chondrocyte C28I2 and mouse macrophage RAW264.7. METHODS The histidine-tagged protein was specifically affinity-purified using Ni-NTA Sefinose resin, and the concentration and purity of the target protein were verified by Coomassie blue staining, BCA method and Western blot. The effects of rhPGRN on the proliferation, autophagy and ERS in C28I2 and RAW264.7 cells were detected by cell counting, Western blot and RT-qPCR. RESULTS The highly purified biologically active human recombinant protein rhPGRN was successfully extracted from the cell line with stable PGRN transfection. rhPGRN promoted the proliferation of the C28I2 cells and RAW264.7 cells, up-regulated the mRNA expression of cell cycle-related molecules (PCNA, cyclin B1 and cyclin D1) and the protein expression of Ki67, and increased the phosphorylation levels of proliferation-related signaling molecules ERK and Akt. Treatment with ERK pathway inhibitor U0126 inhibited rhPGRN-promoted proliferation, autophagy and ERS in the cells. The rhPGRN-induced autophagy of the cells was also inhibited by PI3K/Akt pathway inhibitor 3-methyladenine. The rhPGRN-promoted protein expression of Ki67 was down-regulated by autophagy inhibitor bafilomycin A1 and ERS inhibitor 4-phenylbutyric acid. CONCLUSION These results not only established a method for stable extraction of biologically active high-concentration high-purity recombinant protein rhPGRN, but also confirmed that the biological effect of rhPGRN on promoting cell proliferation was achieved through regulating autophagy and ERS via MAPK and PI3K/Akt pathway. 相似文献
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LI Feng YAO Li ZHANG Xi-hong XIA Yu-hong CHENG Jie LOU Xue-ling JIANG Qiu-hui 《园艺学报》2016,32(12):2251-2255
AIM: To examine the expression of miRNA-22 in the ovarian tissues and the effect of miRNA-22 over-expression on the proliferation, migration and invasion in SKOV-3 cells. METHODS: The expression levels of miRNA-22 in different ovarian tissues and SKOV-3 cells were determined by qPCR. miRNA-22 was over-expressed by transfection of miRNA-22 mimic. The cell viability was examined by CCK-8 assay. The cell migration was measured by wound healing test. The cell invasion was analyzed by Transwell assay. The protein expression levels of VEGF and P53 were determined by Western blot. RESULTS: Compared with the normal ovarian tissue, the expression level of miRNA-22 was remarkably decreased in the ovarian tumor tissues. After transfection with miRNA-22 mimic, the expression level of miRNA-22 in the SKOV-3 cells was significantly increased, while the cell viability, migration and invasion were obviously decreased. Moreover, the protein expression of VEGF and P53 was dramatically inhibited after over-expression of miRNA-22. CONCLUSION: The decreased miRNA-22 expression may be correlated with the development of ovarian can-cer. Over-expression of miRNA-22 decreases the cell viability, migration and invasion by reducing the protein expression of VEGF and P53. 相似文献
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AIM: To investigate the effect of Ikaros isoforms on the proliferation of human ovarian cancer SKOV3 cells. METHODS: Three isoforms of Ikaros, IK1, IK2 and IK6, were transfected into ovarian cancer SKOV3 cells. CCK-8 assay and cell counting were used to detect the effects of Ikaros isoforms on the proliferation of SKOV3 cells. The cell cycle was analyzed by flow cytometry. The cell cycle-related proteins were detected by Western blot. RESULTS: IK1 and IK2 expression inhibited SKOV3 cells proliferation. Flow cytometry analysis indicated that IK1 and IK2 induced SKOV3 cell cycle arrest at the G1 phase. IK6 isoform exerted no obvious effect on the proliferation or cell cycle of SKOV3 cells. Compared with control EV group, IK1 group and IK2 group showed a dramatic elevation in the expression of the cell cycle inhibitor p21, along with a substantial decrease in the expression of the cell cycle inducers cyclin D1 and cyclin D2, which did not change in IK6 group. CONCLUSION: IK1 and IK2 significantly inhibit the proliferation of ovarian cancer SKOV3 cells and induce cell cycle arrest at G1 phase by regulation of cell cycle-related proteins cyclin D1, cyclin D2 and p21, while IK6 isoform exerts no obvious effect on the proliferation and cell cycle of SKOV3 cells. 相似文献
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AIM: To investigate the effect of decitabine (Dacogen, DAC) on the proliferation and differentiation of K562 cells. METHODS: The K562 cells were treated with different concentrations of DAC. The colony formation ability of the cells was detected by the colony formation assay with semi-solid culture. The cell viability was detected with MTT assay. The morphologic features were observed under inverted microscope with Wright's staining. The changes of the cell cycle distribution and the expression of CD11b and CD42b were analyzed with flow cytometry. The protein expression of CDK2, cyclin E1, P27, GATA-1 and PU.1 in the K562 cells was determined by Western blot. RESULTS: DAC significantly decreased the colony number of the cells and cell viability in a dose-dependent manner. The morphological changes of the cells displayed partial differentiation. After treated the K562 cells with DAC for 72 h, the cell proportion in S phase was obviously decreased, while the cell proportion in G2/M phase was obviously increased in a dose-dependent manner. After treated the K562 cells with DAC for 7 d, the percentage of CD11b and CD14 positive cells was further elevated, and the protein expression of P27, GATA-1 and PU.1 was increased. However, the protein expression of CDK2 and cyclin E1 was decreased. CONCLUSION: DAC inhibits the proliferation and induces differentiation of the K562 cells via regulation of cell cycle. 相似文献
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AIM: To explore the influence of long non-coding RNA HOTAIR on the proliferation and apoptosis of acute lymphoblastic leukemia cells via the regulation of glucocorticoid receptor (GR).METHODS: The expression le-vels of HOTAIR and GR mRNA in human bone marrow stromal cell line HS-5 and human acute lymphoblastic leukemia cell lines MOLT-4, CCRF-CEM and CEM-C1 were examined by RT-qPCR. HOTAIR was knocked down by siRNA in acute lymphoblastic leukemia cells. CCK-8 assay was used to assess the cell viability, and the effect of si-HOTAIR on the proli-feration of CEM-C1 cells was evaluated by BrdU method. The effect of si-HOTAIR on apoptosis of CEM-C1 cells was examined by Hoechst 33342 staining and Caspase-Glo® 3/7 assay. Western blot was utilized to examine the protein level of GR.RESULTS: The expression level of HOTAIR in acute lymphoblastic leukemia cells was significantly increased as compared with normal human bone marrow stromal cells (P<0.01). The viability and proliferation of acute lymphoblastic cells was inhibited, the apoptosis was induced, and the anti-proliferation effect of dexamethasone on CEM-C1 cells was enhanced after knockdown of HOTAIR expression (P<0.01). The expression of GR was up-regulated at both mRNA and protein levels (P<0.01).CONCLUSION: Long non-coding RNA HOTAIR may modulate the viability, proliferation and apoptosis of acute lymphoblastic leukemia cells via a GR regulatory way. 相似文献
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AIM: To investigate the characteristic of T-cell acute lymphocytic leukemia 1 (TAL1) gene expression in acute myeloid leukemia (AML) cell lines and in primary AML cells from de novo AML patients with different subtypes. METHODS: Real-time PCR was used to determine the expression of TAL1 mRNA in acute leukemia cell lines (Jurkat, CCRF-CEM, HL-60 and NB4 cell lines) and peripheral blood mononuclear cells from 47 newly diagnosed AML patients. Twelve healthy individuals were served as healthy control group. RESULTS: A significantly increased level in TAL1 mRNA was found in AML cell lines (HL-60 and NB4), T-cell acute lymphacytic leukemia (T-ALL) cell lines (Jurkat, CCRF-CEM) and primary AML cells compared with the healthy controls. Over-expression of TAL1 was found in all detected AML subtypes, the highest level of TAL-1 mRNA was found in AML-M1 and AML-M5 subtype (P<0.05). CONCLUSION: High expression of TAL1 in AML might influence the differentiation and proliferation of myeloid cells, further investigation needs to confirm whether it would be as a biomarker for pathogenesis of AML. 相似文献
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AIM: To investigate the expression of cystic fibrosis transmembrane conductance regulator (CFTR) in acute myeloid leukemia (AML) and its effect on the biological function of human erythroleukemia cell line TF1, and to explore the underlying mechanism. METHODS: The abundance of CFTR in the bone marrow mononuclear cells of patients with AML was measured by real-time PCR. After TF1 cells were incubated with CFTR specific inhibitor CFTRinh-172, cell viability, cell cycle distribution and cell apoptosis were analyzed by CCK-8 assay and flow cytometry. The Wnt signaling pathway-related proteins were determined by Western blot. RESULTS: CFTR was highly expressed in both patients with AML and leukemia cell lines. After incubated with CFTRinh172, the viability of TF1 cells was decreased, the proportion of the cells in G0/G1 phase was increased, while that in S phase declined (P<0.05). Furthermore, the cells treated with CFTRinh-172 exhibited higher apoptotic rate, accompanied with lower protein expression of β-catenin, c-Myc and cyclin D1 (P<0.05). CONCLUSION: CFTR expression is dramatically increased in AML. Inhibition of CFTR restrains the growth and promotes the apoptosis of TF1 cells via classical Wnt signaling pathway. 相似文献
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AIM: To investigate the inhibitory effect of CC-223, an inhibitor of mammalian target of rapamycin (mTOR) kinase, on the viability of human breast cancer cells and its mechanism. METHODS: The inhibitory effect of CC-223 on the viability of MCF-7 cells and MDA-MB-231 cells was measured by CCK-8 assay. The cell cycle distribution of breast cancer cells was examined by flow cytometry. The expression of cell cycle-related proteins and oncoproteins c-Myc and survivin was analyzed by Western blot. RESULTS: CC-223 significantly inhibited the viability of both MCF-7 and MDA-MB-231 cells (P <0.05). CC-223 induced cell cycle arrest in both G1 phase and G2/M phase in the MCF-7 cells (P <0.05). However, low concentration of CC-223 treatment resulted in the accumulation of MDA-MB-231 cell cycle in the G2/M phase, and the cell number in G1 phase was unaffected. Treatment with CC-223 for 24 h clearly inhibited the protein levels of cyclin B1, cyclin D1 and phosphorylated cell division cycle protein 2 in the breast cancer cells (P <0.05). CC-223 suppressed the expression of c-Myc and survivin in both MCF-7 cells and MDA-MB-231 cells (P <0.05). CONCLUSION: CC-223 inhibits cell viability by blocking cell cycle progression and down-regulating expression of c-Myc and survivin in both MCF-7 and MDA-MB-231 cells. 相似文献
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Ursolic acid inhibits migration and invasion of human lung cancer A549 cells by targeting miRNA-133a
AIM: To investigate the effects of ursolic acid (UA) on the migration and invasion of human lung cancer cell line A549, and to explore its mechanism. METHODS: The cell viability was detected by MTT assay. The expression of miRNA-133a was detected in the A549 cells treated with UA by real-time PCR. The miRNA-133a mimics and inhibitor were transfected into the A549 cells, and the transfection efficiency was analyzed by real-time PCR. The cell migratory and invasive abilities were determined by wound healing and Transwell methods, respectively. RESULTS: The viability of the human lung cancer A549 cells was significantly inhibited by UA in a dose-dependent manner (P<0.05). IC50 of UA (24 h) for lung cancer A549 cells was 31.04 μmol/L. UA treatment significantly inhibited the migratory and invasive abilities of A549 cells in a concentration-dependent manner, accompanied by significantly elevation of miRNA-133a expression. The mimics and inhibitor of miRNA-133a significantly upregulated and downregulated the expression of miRNA-133a in the transfected A549 cells, respectively. In addition, the viability of the A549 cells was decreased extremely after tansfected with the miRNA-133a mimics (P<0.01), so did the results of the cell migration and invasion test. The A549 cells tansfected with the miRNA-133a inhibitor showed an opposite changes of the cell viability, migration and invasion. CONCLUSION: UA inhibited the viability, migration and invasion of lung cancer A549 cells by elevating the expression of miRNA-133a. 相似文献
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AIM To investigate the effect of over-expression of BTB and CNC homology 2 （BACH2） on the viability and apoptosis of human acute lymphoblastic leukemia T lymphocytes CCRF-CEM. METHODS CCRF-CEM cells were divided into 3 groups： control group, empty vector group, and BACH2 over-expression group. The BACH2 over-expression vector was transfected into CCRF-CEM cells of BACH2 over-expression group by liposome transfection method. The difference in mRNA expression of BACH2 between CCRF-CEM cells and peripheral blood mononuclear cell （PBMC） was detected by qPCR. CCK8 assay was performed to evaluate the viability of CCRF-CEM cells. Flow cytometry was used to analyzed the apoptosis of CCRF-CEM cells. The protein expression of BACH2 and cyclin D3 in the CCRF-CEM cells was observed by immunofluorescence staining. The protein expression of cyclin D3, Bcl-2 and caspase-3 was determined by Western blot. RESULTS The mRNA expression of BACH2 in CCRF-CEM cells was significantly lower than that in PBMC （P <0.05）. Compared with control group, BACH2 over-expression significantly suppressed the viability,increased the apoptotic rate and caspase-3 expression, and decreased the expression of cyclin D3 and Bcl-2 in CCRF-CEM cells （P <0.05）. CONCLUSION BACH2 expression is decreased in T lymphocytes of human acute lymphoblastic leukemia. Over-expression of BACH2 inhibited the viability of human acute lymphoblastic leukemia T lymphocyte and induced apoptosis. 相似文献
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ZHAO Chao-hui CHEN Xiao-chun JIN Jian-sheng ZHU Yuan-gui SHI Guang-bin ZENG Yu-qi LI Yong-kun PENG Xiao-song 《园艺学报》2005,21(2):238-242
AIM: To investigate the mechanism of cell cycle in aging model induced by tert-butylhydroperoxide(t-BHP).METHODS: From the 30th population doubling(PD), WI-38 cells were exposed for 1 h to t-BHP at every two PDs and four stresses were induced. The cell morphology and ultrastructure, cell cycle assay and β-galactosidase cytochemistry staining were used to evaluate cell senescence. Then the levels of CDK4, CDK2, cyclin D1, cyclin E, p21 and p16 protein were detected by Western blotting.RESULTS: After four times treated with 100 μmol/L t-BHP, the proliferation and division in WI-38 cells were ceased, which showed that cell became larger and flat, the number of secondary lysosome and activity of SA-β-galactosidase were increased. In addition, the percentage of cells in the G1 phase was decreased. Furthermore, it showed that the protein levels of CDK4, CDK2 and cyclin E was reduced and cyclin D1, p21 and p16 was elevated.CONCLUSION: t-BHP induces senescence in WI-38 cells. The possible mechanism is that t-BHP can change the expression of proteins related with cell cycle. 相似文献
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QIN Xiao-ping ZHAN Xiong-yu CHEN Qi-biao HUANG Bao-yuan HUANG Jun ZHUO Yu-min 《园艺学报》2018,34(6):1025-1030
AIM: To observe the effects of the combination of berberin (Ber) and mitomycin C (MMC) on the cell cycle arrest and apoptosis of T24 bladder cancer cells and the underlying mechanisms. METHODS: The T24 cells were exposed to MMC in the presence or absence of difference concentrations of Ber. The viability of the T24 cells was determined by CCK-8 assay. The cell cycle distribution was detected by flow cytometry. The apoptosis was analyzed by flow cytometry with Annexin V-FITC/PI staining, and the protein expression levels of cyclin D1, survivin, CDK2, CDK4, p21 and p27 were determined by Western blot. RESULTS: CCK-8 experiments showed that Ber enhanced the inhibitory effect of MMC on the viability of T24 cells. The results of flow cytometry showed that Ber also enhanced the blockade effect of MMC on T24 cells in G0/G1 phase (P<0.05). Compared with the MMC group, Ber increased the expression of p21 and p27 up-regulated by MMC, and decreased the expression of cynlin D1, CDK2 and CDK4 (P<0.05). Meanwhile, Ber promoted MMC to inhibit the expression of survivin (P<0.05). Ber increased the apoptosis of T24 cells induced by MMC (P<0.05). CONCLUSION: Ber significantly enhances the inhibitory effect of MMC on the viability of T24 cells. The mechanism may be related to up-regulation of p21 and p27, thereby inhibiting the expression of cyclin D1, CDK-2 and CDK-4. At the same time, Ber inhibits the protein expression of survivin, which eventually leads to cell arrest in G0/G1 phase and promotes apoptosis. 相似文献