| 摘要: |
| 目的探索靶向沉默成纤维细胞生长因子受体1（FGFR1）表达对胃癌细胞SGC-7901生长及凋亡的影响。方法通过Westernblotting法研究FGFR1蛋白在胃癌细胞SGC-7901以及人正常胃黏膜上皮细胞GES-1的表达情况；设计合成特异性针对FGFR1基因的小干扰RNA（siRNA）及阴性对照-siRNA（NC-siRNA）；采用瞬时转染法将siRNA转入胃癌细胞SGC-7901中，Westernblotting法检测转染siRNA-FGFR1后对SGC-7901细胞中FGFR1蛋白表达的影响。通过MTT法检测siRNA-FGFR1对SGC-7901细胞增殖抑制的时间-效应关系，以及克隆集落形成实验检测siRNA-FGFR1对SGC-7901细胞克隆集落形成的影响；采用Hoechst染色法和流式细胞术检测siRNA-FGFR1对胃癌细胞凋亡的影响。结果与人正常胃黏膜上皮细胞GES-1相比，胃癌细胞SGC-7901的FGFR1蛋白表达水平明显增高（P＜0.05）；siRNA-FGFR1转染SGC-7901细胞后，FGFR1蛋白表达水平明显下调（P＜0.05）；胃癌细胞增殖生长曲线提示，用siRNA技术沉默胃癌细胞中FGFR1的表达后，相较于空白组和NC-siRNA组，siRNA-FGFR1组细胞增殖明显受抑制，另外胃癌细胞SGC-7901克隆集落形成明显受抑制；流式细胞术结果表明，正常组与NC-siRNA组中胃癌SGC-7901细胞早期凋亡百分比差异无统计学意义（P＞0.05），而siRNA-FGFR1组细胞早期凋亡百分比显著升高（P＜0.01）。结论siRNA-FGFR1可抑制FGFR1蛋白在人胃癌细胞SGC-7901中的表达，并抑制胃癌细胞的生长，促进其凋亡。以FGFR1为靶点的小RNA干扰技术有望成为胃癌靶向治疗的新方法。 |
| 关键词: 胃癌 增殖 凋亡 成纤维细胞生长因子受体1 |
| DOI：10.12056/j.issn.1006-2785.2017.39.22.2016-853 |
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| 基金项目: |
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| Effect of siRNA-mediated FGFR1 gene silencing on growth and apoptosis of gastric cancer cell line SGC-7901 |
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Taizhou Hospital
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| Abstract: |
| Objective To investigate the effect of siRNA-induced FGFR1 gene down-regulation on proliferation and apoptosis of gastric cancer SGC-7901 cells. Methods siRNA-FGFR1 and the negative-control(NC)-siRNA were designed and synthesized, then transfected into SGC-7901 cells. The expression levels of FGFR1 protein was detected by Western blotting analysis. The MTT assay and cell clonogenic assay were used to investigate the inhibitory effect of siRNA-FGFR1 on the viability of gastric cancer SGC-7901 cells. The apoptosis of SGC-7901 cells was detected by flow cytometry with Hoechst staining. Results The expression of FGFR1 in SGC-7901 cells was higher than that in human normal gastric mucosa epithelial GES-1
cells (P＜0.05), and the expression level of FGFR1 in SGC-7901 cells after transfection with siRNA-FGFR1 was down-regulated
(P ＜0.01). The MTT and clonogenic assay showed that the growth of SGC-7901 cells was inhibited after transfecton with siRNA-FGFR1 in comparison to that in NC-siRNA and mock groups. Flow cytometry and Hoechst staining showed that the apoptotic rate was significantly increased in siRNA-FGFR1 transfected cells compared to control cells. Conclusion siRNA-FGFR1 can significantly inhibit FGFR1expression, suppress the growth and induce apoptosis in FGFR1-overexpressing gastric cancer SGC-7901 cells. |
| Key words: Gastric cancer ( GC) Proliferation Apoptosis Fibroblast growth factor receptor 1 ( FGFR1) |
