Bcl-2 siRNA对胃癌细胞凋亡及其紫杉醇敏感性影响研究  

Bcl-2 siRNA increase the sensitivity of gastric cancer cells to paclitaxel treatment by promoting it's apoptosis

作　　者：陈曦[1] 林振兴[2] 林锦娟[2] 陈英玉[2] 余宗阳[1] 欧阳学农[1] 

机构地区：[1]南京军区福州总医院肿瘤科,福建福州350025 [2]福建省血液病研究所.福建医科大学附属协和医院,福建福州350001

出　　处：《中华肿瘤防治杂志》2013年第20期1580-1584,共5页Chinese Journal of Cancer Prevention and Treatment

基　　金：福建省自然科学基金(Z0516080)

摘　　要：目的:研究特异性siRNA对胃癌细胞的抑制作用,以及探讨采用RNA干扰技术下调Bcl-2基因能否增强SGC-7901胃癌细胞对紫杉醇的敏感性。方法:通过脂质体HiperFect将Bcl-2 siRNA转染入胃癌细胞SGC-7901;采用RT-PCR和蛋白印迹法检测Bcl-2 siRNA转染前后胃癌细胞Bcl-2基因表达的变化;用Annexin Ⅴ法及细胞周期分析法检测细胞凋亡;用MTS法观察细胞对药物紫杉醇敏感性的影响。结果:Bcl-2 siRNA组较空白对照组明显抑制Bcl-2 mRNA的表达水平,抑制率为(87.6±5.3)%,P=0.001;Bcl-2 siRNA组较空白对照组明显下调Bcl-2蛋白表达水平,抑制率为(85.7±3.6)%,P=0.003。Bcl-2 siRNA组、紫杉醇组和紫杉醇+Bcl-2 siRNA细胞总凋亡率分别为49.00%、46.10%和66.00%,联合用药组细胞凋亡率最强。Bcl-2siRNA+紫杉醇联合用药对SGC-7901细胞周期分析结果显示,联合用药组凋亡率为48.00%,Bcl-2 siRNA组和紫杉醇组分别为9.03%和21.50%。Bcl-2 siRNA组、阴性对照组和空白对照组的IC50分别(7.25±0.22)、(54.45±0.82)和(68.9±0.91)μg/L,Bcl-2 siRNA转染胃癌细胞SGC-7901对药物紫杉醇更敏感,其IC50值比阴性siRNA转染组降低86.7%,即对紫杉醇敏感性增加7.25倍。结论:靶向Bcl-2 siRNA可明显下调靶基因Bcl-2表达,促进SGC-7901细胞凋亡并能提高细胞对紫杉醇敏感性,为胃癌治疗提供新的策略。OBJECTIVE：To investigate if downregulation of Bcl-2 by siRNA would sensitize gastric carcinoma cell SGC 7901 to paclitaxel. METHODS：BcI-2 siRNA was transfected into human gastric carcinoma cell by cationic liposome HiperFect,The expression of Bcl-2 gene mRNA and its protein in gastric carcinoma cells were evaluated by semi-quantita- tive RT-PCR and Western blot respectively,The cell apoptosis were examined by Annexin V staining and cell cycle assay, The chemosensitivity of transfected cell to paclitaxel was determined by MTS assy. RESULTS：Bcl-2 siRNA significantly downregulated the expression of Bcl 2 gene in human gastric carcinoma cells at both mRNA （87.6±5.3） % （P=0. 001） and protein levels （85. 7±3. 6）% （P=0. 003）. The cellular apoptosis of Bcl-2 siRNA group was 66. 00%, paclitaxel group was 49.00% ,paclitaxel and Bcl-2 siRNA group was 46. 10%. Combination of Bcl-2 siRNA with paclitaxel signifi cantly increased the rate of apoptosis （48.00 %） comparing with siRNA （9.03 % ） or paclitaxel （21. 50 %） treated alone. The IC50 of groups of Bcl-2 siRNA,negative siRNA and untreated cells siRNA were （7.25±0.22）, （54.45±0.82） and （68.9±0.91） . The Bcl-2 siRNA transfected SGC-7901 cells were much more sensitive to the cytetoxic effect of pa- clitaxel with 86.7 % decrease in the value of ICs0 compared to the negative siRNA transfected SGC-7901 cells. Bcl-2 siR- NA increased the sensitivity of SGC-7901 to paclitaxel by sevenfold. CONCLUSION： The effective siRNA targeting Bcl-2 can downregulat Bcl 2 gene expression and enhanc the chemosensitivity of the SC-7901 gastric cancer cells to paclitaxel through promoting aptoposis in SGC-7901 cells,it is expected to provide a novel therapy approach for gastric cancer.

关 键 词：BCL-2 SIRNA 胃肿瘤 药物疗法 细胞凋亡 紫杉酚 药理学 

分 类 号：R735.2[医药卫生—肿瘤]