共查询到20条相似文献，搜索用时 31 毫秒
1.
ZHANG Meng-xia LUO Hong-mei TANG Sheng-song LEI Xiao-yong LONG Zhi-feng ZHANG Xiao-hong WANG Yu-hua 《园艺学报》2005,21(12):2382-2387
AIM: To study the expression and characterization of intracellular macrophage colony-stimulating factor (M-CSF) in human hepatoma cell line, SMMC 7721 cell, and to explore the mechanism by which M-CSF regulates the proliferation of human hepatoma cells. METHODS: The immunohistochemical staining, flow cytometry, antisense technique and Western blotting were used to study the effects and mechanisms of intracellular M-CSF on the proliferation of human hepatoma cells. RESULTS: SMMC 7721 cells highly expressed M-CSF and its receptor. The localization of positive reactions was mainly in cytoplasma and nucleus in SMMC 7721 cells. In cytoplasma and nucleus, one isoforms of M-CSF was found with the molecular weight (MW) of 20 kD, while one type of M-CSFR was discovered with MW of 120 kD. Immunoprecipitation assay showed that these ligands existed in binding with its receptor. Monoclonal antibody (McAb) against M-CSF and antisense oligodeoxynucleotides (ASODN) blocking M-CSF expression inhibited the proliferation of SMMC 7721 cells. McAb and ASODN regulated the expression of cyclin D1/E and p16. Simultaneous administration of both McAb and ASODN inhibited the proliferation of SMMC 7721 cells and modulated the expression of cyclins at greater degrees. CONCLUSION: Our results suggest that an autocrine and an intracrine loop of M-CSF/M-CSFR are present in SMMC 7721 cells. 相似文献
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AIM: To study the effect and mechanism of yeast cytosine deaminase/ thymidine kinase (yCD/TK) double suicide gene driven by alpha fetoprotein (AFP) promoter on hepatocellular carcinoma (HCC) in vitro and in vivo. METHODS: The expression plasmid with yCD/TK double suicide gene, which was driven by AFP promoter, was constructed. HepG2 (AFP positive) and SMMC7721 (AFP negative) human HCC cell lines were both transfected with the above-mentioned expression plasmid through cationic liposome. The cells were treated with 5-fluorocytosine (5-FC) and/or ganciclovir (GCV) at different concentrations. The cell proliferation and cell cycle phase were evaluated by MTT test and flow cytometry respectively. The effect of double suicide gene on HCC xenografts in nude mice was observed through measuring the tumor size and the number of apoptosis cells. RESULTS: The double suicide gene was expressed selectively on HepG2 cells, rather than on SMMC7721 cells. The 5-FC and/or GCV inhibited effectively the proliferation of HepG2 cells in a dose-dependent manner, but had no influence on SMMC7721 cells. The inhibitory effect on HepG2 cells among different treatments was GCV+5-FC>5-FC>GCV. In vivo, the treatments inhibited markedly the growth of HepG2 cell xenografts in nude mice, transfected with yCD/TK gene. More apoptotic cells were found in HepG2 xenografts after the treatment. However, the growth of SMMC7721 cell xenografts could not be inhibited by this double suicide gene therapy, and few apoptotic cells were found. CONCLUSION: yCD/TK double suicide gene driven by AFP promoter has a significant efficacy in treatment of AFP positive HCC. Cell apoptosis may be an important mechanism of yCD/TK double suicide gene-inhibiting the growth of HCC. 相似文献
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AIM:To study the mechanism of growth inhibitory effect of a selective cyclooxygenase-2 (COX-2) inhibitor,NS-398,on cancer cells.METHODS:The esophageal cancer cell line (EC9706),which expresses COX-2 constitutively,and hepatocellular carcinoma cell line (SMMC7721),which expresses no COX-2,were studied.The cell lines were incubated with NS-398 at doses of 10,20,50,100 μmol/L for 24 h,48 h and 72 h.Antiproliferation effect was measured by ［3H］-TdR incorporation.The cell apoptosis were determined by flow cytometry (FCM) and DNA fragmentation analysis.Survivin was detected by immunocytochemical technique.RESULTS:The growth inhibition was induced by NS398 in a dose- and time-dependent manners in both cell lines.FCM analysis revealed a high sub-G1 cell peak in EC9706 group and agarose electrophoresis showed marked apoptosis ladder pattern.However,no apoptosis was observed in SMMC7721 cells treated with NS-398.The difference of apoptosis percentage in EC9706 and SMMC7721 was (45.23±1.08)% and (3.05±0.15)% (P<0.01).After 24 h incubation with NS-398 at concentration of 100 μmol/L,the expression of survivin was markedly reduced in EC9706,no change was observed in SMMC7721.CONCLUSION:NS-398 suppresses cell growth in cancer cell lines by different mechanism.NS-398 suppresses cell growth and increases apoptosis in the cancer cells that expresses COX-2. 相似文献
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LI Jian-jun CHEN Shi-ping GU Mo-fa YANG Wei-min LI Yong-qiang WANG Qi-jing HE Jia PAN Ke ZHAO Jing-jing XIA Jian-chuan 《园艺学报》2012,28(4):638-642
AIM: To evaluate the role of natural killer(NK) cells against a variety of human hepatocellular carcinoma (HCC) cell lines in vitro and in vivo, and to investigate the expression of MHC class I chain-related protein (MIC protein) in these HCC cell lines. METHODS: Peripheral blood mononuclear cells (PBMC) were isolated from 50 mL peripheral blood donored by a healthy volunteer and cultured in the NK cell kit followed by amplification and cytokine activation. The release of lactate dehydrogenase (LDH) was detected by lysis experiment. Animal experiments were performed following vaccination with HCC cell lines (BEL7402, HepG2 and SMMC7721) in BALB/c-nu/nu mice. The diameters of the tumors were measured and the growth curves of difference cell lines were delineated. Three weeks later, these mice were sacrificed and the weight of the transplanted tumors was also detected. Furthermore, the expression of MIC protein was determined. RESULTS: The obvious differences of lysis effects of NK cells on different cell lines were observed, in which the lysis effect of NK cells on K562 cells was the strongest, the effect on BEL7402 cells was in the middle and the effect on SMMC7721 cells was the weakest. In animal experiments, NK cells also showed obvious and different restraining effects on a variety of HCC cell line-transplanted tumors in nude mice. The tumor inhibitory rates of NK cells were 43.5% in BEL-7402 cell-transplanted tumor, 40.7% in HepG2 cell-transplanted tumor and 36.0% in SMMC-7721 cell-transplanted tumor. The protein expression of MIC was 48.7%, 32.8% and 0.9% in the 3 cell lines,respectively. CONCLUSION: The lysis effects of NK cells on a variety of human HCC cell lines are different. The expression of MIC in the tumor cells may play distinct role in evaluating these differences. 相似文献
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AIM: To explore the effects of the new drug of sulfonylurea (1-{4-［2-(3-ethyl- 4-methyl-2-oxo-3-pyrroline-1- carboxamido)ethyl］-phenylsulfonyl}-3-(1, 4-tetramethylene)-urea, BGW) on the glucose uptake and the activation of Akt/PKB in SMMC7721 cells. METHODS: Cultured SMMC7721 cells were divided into control group, glibenclamide group, insulin group, BGW group and BGW+insulin group. Scintillation was used to detect the glucose uptake in SMM7721 cells. The activation of Akt/PKB was tested by Western blotting. RESULTS: Compared to control cells, gibenclamide, insulin, BGW and BGW+insulin significantly increased the glucose uptake (P<0.01), and increased by 22%, 108%, 60% and 400% in SMMC7721 cells, respectively. The new drug of sulfonylurea increased the insulin-stimulated glucose uptake (P<0.01) in SMMC7721 cells. The BGW-stimulated glucose uptake was higher than that with the glibenclamide-stimulated, and increased the insulin-stimulated glucose uptake (P<0.01). The sulfonylurea significantly enhanced the activity of Akt in SMMC7721 cells. CONCLUSION: The new drug of sulfonylurea (BGW) stimulates glucose uptake, increases the insulin-stimulated glucose uptake in SMMC7721 cells, and is a potential anti-hyperglycemic agent. 相似文献
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AIM: To investigate the effects of recombinant human endostatin (Endostar) on proliferation, cell cycle constitution and regulation of related protein expression in RPMI 8226 cells. METHODS: CCK-8 assay was used to explore the effect of Endostar on the proliferation of RPMI 8226 cells. The action of Endostar on apoptosis and cell cycle distribution were determined by flow cytometry. The expression of Bcl-2 and caspase-3 was evaluated by Western blotting analysis. The production of vascular cell adhesion molecule 1 (VCAM-1), interleukin 6 (IL-6) and vascular endothelial growth factor (VEGF) in response to Endostar was measured by real-time PCR and Western blotting. The influence of Endostar on the secretion of IL-6 and VEGF in the culture supernatants was detected by ELISA. RESULTS: Endostar inhibited the proliferation of RPMI 8226 cells with the highest inhibitory rate of 59.5±5.6% at dose of 250 mg/L, which was associated with slight up-regulation in the proportion of G1 phase. No significant modulation of apoptotic rate was observed with Endostar, as well as the expression of Bcl-2 or caspase-3. Endostar declined the expression of VCAM-1 and also attenuated the production and secretion of IL-6 and VEGF in RPMI 8226 cells. CONCLUSION: Endostar suppresses the proliferation of RPMI 8226 cells by enhancing the proportion of G1 phase and decreasing VCAM-1 expression and self-secretion of IL-6 and VEGF. No apoptosis of RPMI 8226 cells is induced by Endostar. 相似文献
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AIM: To investigate inhibition of K562 cell growth by antisense drug targeted VEGF mRNA. METHODS: X7, 20-mer antisense sequences were selected, synthesized and modified with phosphorothioate. The drug was transfected into K562 cells in the present of lipofection. Cell growth was assayed by trypan blue dye exclusion assay and MTT. The level of VEGF protein in the media was determined by ELISA. The morphology of apoptotic cells were observed by Giemsa staining, and the propotion of apoptotic cells was detected by flow cytometry. RESULTS: The antisense drug inhibited growth of K562 and downregulated expression of VEGF protein significantly, compared with Scrambed control group and showed dose-dependent relation. Signs of apoptosis of K562 cells were not observed. CONCLUSION: Inhibition of K562 cell proliferation, but not cells apoptosis induction is the mechanism of inhibing growth of K562 cells by antisense drug targeted VEGF mRNA. At same time, VEGF has function of promoting K562 cell proliferation, and VEGF mRNA may be a new target attached by drugs. 相似文献
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BAI Hui-li LI Bao-lin HE Fang ZHANG Ru-yi YAN Shu-juan LIU Chen YANG Dan-dan SHI Qiong 《园艺学报》2013,29(11):1921-1927
AIM: To investigate the effects of marrow stromal HS-5 cells on hepatocellular carcinoma SMMC-7721 cells in the tumor microenvironment. METHODS: The effects of HS-5 cell-conditioned medium (HS-5-CM) on the proliferation, migration and invasion abilities of SMMC-7721 cells were detected by MTT, wound-healing and Transwell assays. After co-culture of SMMC-7721 cells with HS-5 cells in the Transwell chamber, the expression of chemokine CCL5 and its receptor CCR5 at mRNA and protein levels in SMMC-7721 cells was examined by quantitative real-time PCR (qRT-PCR), ELISA or Western blotting. Akt and p-Akt473 protein levels in SMMC-7721 cells treated with PI3K inhibitor LY294002 were observed by Western blotting. RESULTS: HS-5-CM promoted the proliferation, migration and invasion abilities of SMMC-7721 cells. The expression of CCL5 and CCR5 at mRNA and protein levels in SMMC-7721 cells was increased after co-cultured with HS-5 cells. PI3K inhibitor LY294002 inhibited the activation of PI3K-Akt signaling pathway and the secretion of CCL5 in SMMC-7721 cells after co-cultured with HS-5 cells. CONCLUSION: HS-5 cells significantly promote the proliferation, migration and invasion abilities of SMMC-7721 cells. Co-culture of SMMC-7721 cells with HS-5 cells activates PI3K-Akt signaling pathway to increase the secretion of CCL5 in SMMC-7721 cells. 相似文献
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DAI Hong-liang GE Shu-qing CHU Ming-hui LIANG Chun-guang WANG Yue JIA Gui-zhi 《园艺学报》2016,32(3):464-469
AIM: To investigate the effect of genistein on the proliferation of human oral cancer TCA8113 cells and to explore the underlying mechanisms.METHODS: The cell proliferation was examined by MTT assay, cell counting and colony formation assay. Western blotting was employed to examine the protein levels of vascular endothelial growth factor(VEGF), extracellular signal-regulated kinase(ERK) and p-ERK. RESULTS: Genistein significantly inhibited the proliferation of TCA8113 cells in a concentration-dependent fashion. Moreover, genistein dose-dependently decreased the protein levels of VEGF, ERK and p-ERK. The expression of VEGF was also blunted by U0126, a specific inhibitor of ERK. U0126 and axitinib, a VEGF receptor antagonist, both significantly inhibited the proliferation of TCA8113 cells. CONCLUSION: Genistein inhibits the proliferation of TCA8113 cells, which may be related to its inhibitory effect on ERK expression and activation, thus subsequently decreasing the expression of VEGF. 相似文献
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AIM: To observe the influence of high expression of miR-15a-5p on the proliferation and migration of human hepatocellular carcinoma SMMC-7721 cells.METHODS: The miR-15a-5p oligonucleotide, which was reconstructed with additional restriction sites of EcoR Ⅰ and Hind Ⅲ, was chemically synthesized and confirmed by sequencing. The miR-15a-5p eukaryotic expression system was constructed by pcDNA6.2-GW/Em-GFP-pre-miR-15a-5p plasmid. The miR-15a-5p was transfected into the SMMC-7721 cells transiently by plasmid, and quantified by quantitative real-time PCR at the mRNA level. The cell viability was measured by CCK-8 assay, and the living cell counting was performed by the method of Trypan blue exclusion. The migration ability of the SMMC-7721 cells with high expression of miR-15a-5p was detected by wound healing test.RESULTS: The sequence of miR-15a-5p oligonucleotide 100% matched the designed sequence. Compared with control group, the miR-15a-5p expression was increased significantly (P<0.05). The viability, the living cell number and the migration ability of the SMMC-7721 cells were decreased in high expression of miR-15a-5p group with statistically significant difference (P<0.05).CONCLUSION: The abilities of proliferation and migration in human hepatocellular carcinoma SMMC-7721 cells are decreased by high expression of miR-15a-5p. 相似文献
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AIM:The effective antisense sequences targeted VEGF mRNA with computer software would be screened and designed, and effect of them on growth K562 cells and protein expression of VEGF were studied with experiments.METHODS:Seven antisense sequences were selected and synthesized, which consisted of 18-20 deoxynucleotide acid and were modified with phosphorothioate, according to principle of low free energy of overall △G37 Overall. Cell growth was assayed by trypan blue dye exclusion assay and level of VEGF protien in the media was determined by ELISA.RESULTS:Six of seven sequences were capable of inhibing growth of K562 cells and downregulating the VEGF protein expression significantly, compared with Scrambed control group. It was found that there was a close correlation between low level of overall △G37 and antisense effectiveness (r=0.887,P<0.01).CONCLUSION:VEGF mRNA antisense oliogdeoxynucleotides, which were designed by computer software of RNAstructure, were able to inhibit growth of K562 and its protein expression. The VEGF mRNA may be new target attached by drugs. At same time, the computer aided design is useful methods to obtain the effective antisense. 相似文献
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AIM: To investigate the effects of 2-deoxy-D-glucose (2-DG) or oxaliplatin (L-OHP) alone and the combination of both on the proliferation and apoptosis of hepatoma cell line SMMC-7721 in vitro. METHODS: Human hepatoma cell line SMMC-7721 was treated with 2-DG or L-OHP alone, or both. The inhibitory effect on the proliferation of SMMC-7721 cells was estimated by MTT method. The q value, which represents synergistic effect, was determined. Apoptotic rate and cell cycle were assayed by flow cytometry. The activity of caspase-3 was detected by a caspase-3 activity assay kit. RESULTS: 2-DG or L-OHP at different concentrations inhibited the growth of SMMC-7721 cells obviously and the inhibitory effect on SMMC-7721 cell growth strongly depended on the exposure time and dose. When the 2 drugs worked together, the inhibitory effect was improved (P<0.05). 2-DG induced cell apoptosis and arrested the cells at G2/M phase. When combined with L-OHP,the 2 drugs induced more severe apoptosis and arrested the cells at S and G2/M phase. Meanwhile, the activity of caspase-3 increased when the 2 drugs used together. CONCLUSION: 2-DG inhibits the growth of hepatoma cell line SMMC-7721. The combination of 2-DG and L-OHP improves the ability of L-OHP to attack the tumor cells. The mechanism might be related to increasing the activity of caspase-3. 相似文献
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XIE Xiao-bin LONG Jie ZHANG Hui-yu WANG Hong-yan XU Ruo-bing LI Shu-hua ZHANG Ya-jie 《园艺学报》2008,24(4):650-654
AIM: To study the inhibitory effect of VEGF-C/Flt-4 system on lymphangiogenesis and lymphatic metastasis of breast cancer. METHODS: Lymphatic endothelial cells (LEC) were cultured in vitro, the effects of VEGF-C and anti-Flt-4 antibody on the proliferation of treated cells were observed. The antisense oligodeoxynucleotides (ASODN) targeting VEGF-C was designed and its effect on VEGF-C gene expression in vitro experiments was observed. The nude mice transplantation tumor model was made and the effects of VEGF-C ASODN on lymphangiogenesis and metastasis in the model were determined. RESULTS: The supernatant of cultured PC3 cells promoted LEC proliferation obviously while the cells treated with anti-Flt-4 antibody were obviously decreased whenever cell counting. The mRNA and protein expression of VEGF-C in MCF-7 cells treated with ASODN were significantly lower than that in control groups in vitro. In vivo ASODN also significantly reduced the VEGF-C mRNA expression detected by RT-PCR. The result of 5-Nase-ALPase enzyme -histochemistry showed that ASODN had obvious inhibitory effect on tumor lymphangiogenesis. Tumor growth velocity in ASODN group was much slower than that in control group. ASODN also inhibited tumor volume and lymphatic metastasis. CONCLUSION: The strong relationships between VEGF-C/Flt-4 system and lymphangiogenesis and lymphatic metastasis of breast cancer have been observed. If the expression of Flt-4 is blocked, the proliferation of LEC induced by tumor cells can be blocked in some degree. ASODN inhibits tumor lymphangiogenesis and lymphatic metastasis by down-regulating VEGF-C expressions. 相似文献
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AIM To investigate the effect of ligustilide on human hemangioendothelial cells (HemECs) and to analyze its mechanism. METHODS The effect of ligustilide at different concentrations on the viability of HemECs was measured by CCK-8 assay. The HemECs were divided into control group and ligustilide （10, 25 and 50 μmol/L） treatment groups, and the proliferation of HemECs was detected by EdU staining. The effects of ligustilide on the angiogenesis of HemECs was tested by microtubule formation experiment. The protein expression of vascular endothelial growth factor (VEGF) and epithelial-mesenchymal transition (EMT)-related markers in HemECs cells was determined by Western blot. RESULTS There was no significant difference in the viability of the cells treated with ligustilide at the concentrations between 0.1~50 μmol/L compared with control cells. Compared with control group, ligustilide at 25 and 50 μmol/L significantly reduced the number of EdU-positive cells and microtubule-like structures (P <0.05), reduced the protein expression level of VEGF (P <0.05), increased the protein expression of E-cadherin, and decreased the protein expression of vimentin and β-catenin (P <0.05). Compared with control group, the expression of VEGF and vimentin was significantly up-regulated, and the protein expression of E-cadherin was significantly down-regulated in VEGF overexpression group (P <0.05). Compared with VEGF overexpression group, the expression of VEGF and vimentin in 50 μmol/L ligustilide-treated VEGF-overexpressing cells were significantly reduced (P <0.05), and the protein expression of E-cadherin was significantly increased (P <0.05). CONCLUSION Ligulide inhibits the proliferation of HemECs, and also inhibits the angiogenesis and EMT process of HemECs by reducing the level of VEGF. 相似文献
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AIM:To investigate the antitumor activity and mechanism of Paecilomyces tenuipes polysaccharide(PTPS).METHODS:PTPS-I was obtained by water extraction and alcohol precipitation, and purified by DEAE-cellulose and Sephadex G-100 chromatography. Human erythroleukemia cell line K562, laryngocarcinoma cell line Hep2 and hepatic carcinoma cell line SMMC-7721were co-cultured with PTPS-I or the conditioned medium which prepared with PTPS-I-stimulated human mononuclear cells (PTPS-I-MNC-CM), and the proliferation of tumor cells was determined. The cell counting kit-8 (CCK-8) was used to determine the proliferation of MNCs. The FQ-RT-PCR was applied to investigate the expression of TNF-α and IL-6 mRNA in MNCs. RESULTS:PTPS-I-MNC-CM inhibited the proliferation of K562, Hep2 and SMMC-7721 cells in vitro (P<0.01). Cytotoxicity of PTPS-I against K562, Hep2 and SMMC-7721 cells was not observed (P<0.01). PTPS-I stimulated the proliferation of MNCs (P<0.01) and significantly enhanced the expression of TNF-α and IL-6 mRNA in MNCs (P<0.05, P<0.01). CONCLUSION:The results suggest that PTPS-I is an immunomodulator and its antitumoral activity is through the immunomodulatory mechanism rather than the direct cytotoxicity against tumor cells. 相似文献
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