小干扰 RNA 沉默 Bcl-2 表达对胃癌 BGC823 细胞放射敏感性的影响。

Effect of silencing Bcl-2 expression by small interfering RNA on radiosensitivity of gastric cancer BGC823 cells.

机构信息

Department of General Surgery, Zhengzhou People's Hospital, Zhengzhou 450001, China.

出版信息

Asian Pac J Trop Med. 2013 Jan;6(1):49-52. doi: 10.1016/S1995-7645(12)60199-0.

DOI:10.1016/S1995-7645(12)60199-0

PMID:23317885

Abstract

OBJECTIVE

To explore the influence of silencing Bcl-2 expression by small interfering RNA (siRNA) on Bcl-2 protein expression, cell apoptosis rate and radiosensitivity of gastric cancer BGC823 cells.

METHODS

siRNA segment for Bcl-2 gene was designed and synthesized, then was induced into gastric cancer BGC 823 cells by liposome transfection. Bcl-2 protein expression was detected by Western Blotting. After X radiation, flow cytometry and clone forming assay were used to determine the effects of RNA interference on BGC823 cell apoptosis rate and radiosensitivity.

RESULT

After the transfection of Bcl-2 siRNA, the positive expression rate of Bcl-2 protein in BGC823 cells was (35.45±2.35)%. Compared with the control group and negative siRNA transfection group, the rate was significantly decreased (P<0.01). The apoptosis rate of BGC823-RNAi cell was (10.81±0.91)%, which was significantly higher than the control group and negative siRNA transfection group (P<0.01). After 48h X radiation, the apoptosis rate of BGC823-RNAi was (28.91±1.40)%, which was significantly higher than the control group and the group without radiation (P<0.01). During clone forming assay D(0), D(q) and SF(2) values in Bcl-2 siRNA1 transfection group were all lower than those in the control group. The radiosensitivity ratio was 1.28 (the ratio of D(0)) and 1.60 (the ratio of D(q)).

CONCLUSIONS

Specific siRNA of Bcl-2 gene can effectively inhibit the expression of Bcl-2 gene, enhance the radiosensitivity and apoptosis of gastric cancer BGC823 cells, having good clinical application perspective.

摘要

目的

探讨小干扰 RNA（siRNA）沉默 Bcl-2 表达对胃癌 BGC823 细胞 Bcl-2 蛋白表达、细胞凋亡率和放射敏感性的影响。

方法

设计并合成 Bcl-2 基因的 siRNA 片段，然后通过脂质体转染将其导入胃癌 BGC823 细胞。采用 Western Blotting 检测 Bcl-2 蛋白表达。X 射线照射后，采用流式细胞术和克隆形成实验检测 RNA 干扰对 BGC823 细胞凋亡率和放射敏感性的影响。

结果

转染 Bcl-2 siRNA 后，BGC823 细胞中 Bcl-2 蛋白的阳性表达率为（35.45±2.35）%，明显低于对照组和阴性 siRNA 转染组（P<0.01）。BGC823-RNAi 细胞的凋亡率为（10.81±0.91）%，明显高于对照组和阴性 siRNA 转染组（P<0.01）。48 h X 射线照射后，BGC823-RNAi 组的凋亡率为（28.91±1.40）%，明显高于对照组和未照射组（P<0.01）。在克隆形成实验中，Bcl-2 siRNA1 转染组的 D（0）、D（q）和 SF（2）值均低于对照组。放射敏感性比为 1.28（D（0）比值）和 1.60（D（q）比值）。