RNAi 介导的 ezrin 基因沉默逆转人胃癌细胞系 SGC-7901 的恶性行为。

RNAi-mediated silencing of ezrin gene reverses malignant behavior of human gastric cancer cell line SGC-7901.

机构信息

Department of Gastroenterology, Sixth People's Hospital Affiliated to Shanghai Jiaotong University, Shanghai, China.

出版信息

J Dig Dis. 2009 Nov;10(4):258-64. doi: 10.1111/j.1751-2980.2009.00394.x.

DOI:10.1111/j.1751-2980.2009.00394.x

PMID:19906104

Abstract

OBJECTIVE

To investigate the effects of ezrin targeting gene of RNA interference (RNAi) on human gastric cancer cell line SGC-7901 in vitro.

METHODS

The highly metastatic human gastric cancer cell line SGC-7901 transfected with a small interfering (siRNA) lentivirus vector was selected for this research study. Expressions of ezrin mRNA and ezrin protein in the SGC-7901 cells were detected using RT-PCR and Western blot. Cell apoptosis was observed using flow cytometry. Transwell invasion and the cell adhesion test were used to verify the effect of RNAi on ezrin expression in the human gastric cancer cell line SGC-7901 in vitro.

RESULTS

Ezrin gene targeting via a RNAi-mediated lentivirus vector had obvious inhibitory effects on ezrin expression in the human gastric cancer cell line SGC-7901. The results of the RT-PCR show the obvious inhibition of ezrin mRNA expression in Eai and Ebi groups (0.22 +/- 0.01 vs 0.95 +/- 0.04, P < 0.05; 0.31 +/- 0.01 vs. 0.95 +/- 0.04, P < 0.05). Western blot analysis revealed a 72.35 +/- 3.74% reduction of the ezrin protein level after interference with the ezrin targeting gene. Moreover, the inhibition of ezrin expression clearly inhibited SGC-7901 cell migration and invasion, and improved cell adhesion as well as increased sensitivity to camptothecin-induced apoptosis.

CONCLUSION

Ezrin gene targeting by RNAi can inhibit the metastatic growth and migration of SGC-7901 human gastric cancer cells.

摘要

目的

研究 RNAi 靶向 ezrin 基因对体外人胃癌细胞系 SGC-7901 的影响。

方法

本研究选择转染小干扰（siRNA）慢病毒载体的高转移性人胃癌细胞系 SGC-7901。采用 RT-PCR 和 Western blot 检测 SGC-7901 细胞中 ezrin mRNA 和 ezrin 蛋白的表达。采用流式细胞术观察细胞凋亡。采用 Transwell 侵袭和细胞黏附试验验证 RNAi 对体外人胃癌细胞系 SGC-7901 中 ezrin 表达的影响。

结果

RNAi 介导的慢病毒载体靶向 ezrin 基因对人胃癌细胞系 SGC-7901 中 ezrin 表达具有明显的抑制作用。RT-PCR 结果显示 Eai 和 Ebi 组 ezrin mRNA 表达明显抑制（0.22±0.01 比 0.95±0.04，P<0.05；0.31±0.01 比 0.95±0.04，P<0.05）。Western blot 分析显示，干扰 ezrin 靶向基因后，ezrin 蛋白水平降低 72.35±3.74%。此外，抑制 ezrin 表达明显抑制 SGC-7901 细胞迁移和侵袭，提高细胞黏附性，并增加喜树碱诱导的细胞凋亡敏感性。