RNA干扰survivin基因表达对肝癌细胞放射敏感性影响  

EFFECT OF RNA INTERFERENCE TARGETING SURVIVIN GENE ON RADIOSENSITIVITY OF HEPATOCARCINOMA CELLS

作　　者：肖超[1] 雷炜[1] 张碧媛[1] 姚如永[2] 

机构地区：[1]青岛大学医学院附属医院肿瘤科,山东青岛266003 [2]青岛大学医学院附属医院中心实验室,山东青岛266003

出　　处：《齐鲁医学杂志》2013年第1期5-7,10,共4页Medical Journal of Qilu

摘　　要：目的探讨RNA干扰survivin基因表达对肝癌细胞放射敏感性的影响。方法构建针对人肝癌HepG2细胞survivin基因靶序列的shRNA真核表达载体pshRNA-survivin-387。设置空白对照组(不加载体)、阴性对照组(加入pshRNA-survivin-NC)及siRNA转染组(加入pshRNA-survivin-387),用脂质体2000转染HepG2细胞,采用逆转录聚合酶链反应技术检测各组survivin基因mRNA表达水平。分别设置空白对照组、阴性对照组、siRNA转染组、X线照射组、siRNA转染+X线照射组,收集各组细胞,流式细胞仪检测细胞凋亡率,MTT法测定细胞存活分数。结果 siRNA转染组survivin基因mRNA表达水平明显低于空白对照组及阴性对照组,差异有显著性(F=218.93,q=17.73、18.66,P<0.05)。siRNA转染+X线照射组细胞凋亡率明显高于其他各组,差异有显著性(F=1 421.83,q=13.81～57.48,P<0.05);siRNA转染组及X线照射组与空白组、阴性组比较,细胞凋亡率也明显提高(q=17.55～39.10,P<0.05)。siRNA转染+X线照射组细胞存活分数显著低于其他各组(F=2870.58,q=15.32～72.51,P<0.05);siRNA转染组及单纯X线照射组细胞存活分数与空白对照组、阴性对照组比较也明显降低(q=11.14～73.90,P<0.05)。结论 pshRNA-survivin-387转染能抑制survivin基因表达,诱导肝癌细胞凋亡,增强肝癌细胞的放射敏感性。Objective To study the effect of RNA interference targeting survivin gene on radiosensitivity of hepatocarci- noma cells. Methods The shRNA vectors pshRNA-survivin 387 targeting survivin gene sequence were constructed. This study included blank control group （no vectors added）, negative control group （psh-RNA-surviviwNC added） and siRNA group （psh- RNA survivin-387 added）. HepG2 cells were transfected by liposomes 2000, the expression of survivin mRNA was detected by RT PCR. Blank control group, negative control group, siRNA group, irradiation group, and siRNA＋ irradiation group were es- tablished. The apoptosis in each group was detected by flow cytometry, and cell survival fraction by MTT test. Results The expression of survivin mRNA siRNA group was markedly lower than that in the blank and negative-control groups, the difference being significant （F=218.93;q= 17.73,18.66;P〈0.05）. The rate of apoptosis in siRNA -- irradiation group was significantly higher than that of other groups （F=I 421.83;q= 13.81-57.48;P〈0.05）, that in siRNA group and irradiation group was also higher than that in the blank and negative-control groups （q= 17.55 39.10,P〈0.05）. The survival fraction of siRNA ＋ irradia- tion group was significantly lower than that of other groups （F=2 870.58,q=15.32-72.51,P〈0.05）, that of siRNA group and irradiation group was obviously lower that that of the blank and negative control （q = 11.14- 73.90, P〈0.0.）. Conclusion Transfection of the pshRNA survivin 387 to HepG2 cell can inhibit the expression of survivin gene, induce the apoptosis of liver cancer cells,and increase the sensitivity of the cells to radiotherapy.

关 键 词：肝肿瘤 RNA干扰 SURVIVIN 放射疗法 辐射耐受性 

分 类 号：R735.7[医药卫生—肿瘤]