| 张 乐,白月奎,刘凯东.c-Raf基因对大肠癌细胞生长的作用及其分子机制研究[J].肿瘤学杂志,2019,25(4):293-299. |
| c-Raf基因对大肠癌细胞生长的作用及其分子机制研究 |
| Effect of C-Raf Gene on Growth of Colorectal Cancer Cells and Its Molecular Mechanism |
| 投稿时间：2017-09-11 |
| DOI：10.11735/j.issn.1671-170X.2019.04.B002 |
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| 中文关键词: c-Raf基因 大肠肿瘤 分子机制 |
| 英文关键词:C-Raf gene colorectal cancer molecular mechanism |
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| 摘要点击次数: 2042 |
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| 中文摘要: |
| 摘 要：［目的］ 通过转染siRNA沉默c-Raf基因以探讨其在大肠癌细胞生长中的作用及其相关分子机制。［方法］通过阳离子脂质体介导的方法将siRNA转染入大肠癌细胞HCT116和SW620，经Western blot验证siRNA的干扰效果。经MTT法、Transwell实验、细胞克隆形成实验和流式细胞术研究下调c-Raf基因对HCT116和SW620增殖、迁移和相关细胞活力的影响。采用Western blot法检测转染siRNA后相关细胞周期蛋白水平。［结果］ HCT116和SW620转染后细胞活力明显下降，转染siRNA 48h后HCT116与SW620细胞的抑制率分别为34%、28%。流式细胞术发现HCT116和SW620在c-Raf基因下调后，较多细胞滞留在G1期(P<0.05)。Western blot实验发现转染siRNA后细胞p-Cdc2、E2F1、CyclinD1表达水平均有下降(P<0.05)。siRNA转染HCT116 和SW620细胞后，细胞凋亡比例分别为9.68%±2.37%、7.29%±1.68%，均高于对照组(P<0.05)。siRNA转染HCT116 和SW620细胞后Caspase-3相对水平分别为0.57±0.11、0.47±0.09，Bcl-2相对水平分别为0.16±0.05、0.23±0.04，与对照组相比差异均有统计学意义(P<0.05)。siRNA转染HCT116和SW620细胞后N-cadherin相对水平分别为0.24±0.07、0.22±0.04，E-cadherin相对水平分别为0.47±0.12、0.58±0.13，与对照组相比差异均有统计学意义(P<0.05)。［结论］通过下调c-Raf基因的表达可将大肠癌细胞阻滞在G1期，促使细胞发生凋亡，同时抑制大肠癌细胞发生EMT转化。 |
| 英文摘要: |
| Abstract：［Objective］ To investigate the role of c-Raf gene in the growth of colorectal cancer cells and its molecular mechanism. ［Methods］ SiRNA was transfected into colorectal cancer HCT116 and SW620 cells with cationic liposome method，and the interference effect of siRNA was verified by Western blot. The effects of c-Raf gene on proliferation，migration，related cell viability of HCT116 and SW620 were studied by MTT assay，Transwell assay，cell clone formation assay and flow cytometry，respectively. The expression levels of p-Cdc2，E2F1 and CyclinD1 were detected by Western blot. ［Results］ The viability of HCT116 and SW620 decreased significantly after the siRNA transfection(P<0.05). And the inhibition rate of HCT116 and SW620 cells was 34% and 28% respectively after transfection of siRNA for 48h. The down-regulated c-Raf gene induced G1-phase arrest in HCT116 and SW620 cells. The expression of p-Cdc2，E2F1 and CyclinD1 decreased after siRNA transfection(P<0.05). After siRNA transfection，the percentage of apoptosis in HCT116 and SW620 cells was 9.68%±2.37% and 7.29%±1.68% respectively，which was significantly higher than that in the control group(P<0.05). After siRNA transfection the relative level of Caspase-3 in HCT116 and SW620 cells was 0.57±0.11 and 0.47±0.09，the relative level of Bcl-2 was 0.16±0.05 and 0.23±0.04，and there was significant difference with the control group(P<0.05)，After siRNA transfection the relative level of N-cadherin was 0.24±0.07and 0.22±0.04，the relative level of E-cadherin was 0.47±0.12 and 0.58±0.13 in HCT116 and SW620 cells，and the difference with the control group was significant(P<0.05). ［Conclusion］ The expression of c-Raf gene can arrest colorectal cancer cells in G1 stage，induce cell apoptosis and inhibit the EMT transformation of colorectal cancer cells. |
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