小干扰RNA介导雌激素β基因沉默对肺腺癌LTEP-a2细胞凋亡的影响  

Effects of estrogen receptor β gene silencing mediated by small interference RNA on apoptosis of human lung cancer LTEP-a2 cells

作　　者：卢珠明[1] 伍硕允[1] 王庆涛 李宇明 叶敏[1] 林志潮[1] 沈涛[1] 段楚骁 张东熙 

机构地区：[1]江门市中心医院中山大学附属江门医院胸外科,529030 [2]江门市中心医院中山大学附属江门医院普外科,529030

出　　处：《中华实验外科杂志》2017年第10期1711-1714,共4页Chinese Journal of Experimental Surgery

基　　金：广东省科技计划项目（20128061700064、2017ZC0468）;江门市科技计划项目（2012-58）

摘　　要：目的观察小干扰RNA（siRNA）介导雌激素β（ERβ）基因沉默对ERβ高表达的肺腺癌LTEP-a2细胞凋亡作用及17-β雌二醇敏感性。方法合成针对ERβ的siRNA，对生长期的ERβ高表达的LTEP-a2细胞进行转染，48 h后，收集细胞随机分为阴性对照组、空白对照组和转染组。反转录-聚合酶链反应（RT-PCR）检测ERβ mRNA的表达，Western blot法检测ERβ蛋白的表达。噻唑蓝（MTT）比色法检测17-β雌二醇培养的转染细胞的存活，计算各组细胞的半数抑制率（IC50），流式细胞仪检测转染细胞的凋亡。结果阴性对照组、空白对照组和转染组ERβ mRNA的表达强度分别为0.32±0.06、0.28±0.05和0.05±0.02。转染siRNA组ERβ mRNA表达明显低于阴性对照组（P＝0.032）及空白对照组（P＝0.015）。阴性对照组、空白对照组和转染组ERβ蛋白的表达强度分别为0.18±0.04、0.28±0.03和0.06±0.01。转染siRNA组ERβ蛋白表达低于阴性对照组（P＝0.038）及空白对照组（P＝0.000）；不同浓度的17-β雌二醇干预后，ERβ-siRNA转染组细胞与阴性对照组及空白对照组比较存活率显著下降。阴性对照组、空白对照组和转染组IC50值分别为（28.34±4.32）%、（27.66±3.04）%和（8.34±1.22）%，雌二醇干预对LTEP-a2细胞生长与阴性对照组及空白对照组比较，转染组生长明显抑制；转染组、阴性对照组与空白对照组细胞凋亡率分别为（35.68±1.58）%、（3.55±0.33）%和（1.30±1.45）%，转染siRNA组在雌二醇干预后LTEP-a2细胞的凋亡率显著增加，转染组与阴性对照组与空白对照组比较，差异有统计学意义。结论siRNA介导的Erβ基因沉默可以抑制雌激素干预后肺癌细胞的增殖siRNA介导的Erβ基因沉默，可以增加雌激素干预后肺癌细胞的凋亡，Erβ可能在筛选女性性激素依赖性非小细胞肺癌中发挥重要作用。ObjectiveTo investigate the effect of estrogen receptor β （ERβ） gene silencing mediated by small interference RNA （siRNA） on apoptosis of human lung cancer LTEP-a2 cells overexpressing ERβ and the sensitivity to 17β-estradiol. MethodssiRNA was designed and synthesized as expression vector and transfected into human lung cancer LTEP-a2 cells. After 48 h, the collected cells were randomly divided into the negative control group, the blank control group and the transfected group. Transcription-polymerase chain reaction （RT-PCR） was used to detect ERβ mRNA, and Western blotting was used to detect the expression of ERβ protein. Cellular growth was tested by methylthiazoltetrazolium colorimetry （MTT） assay and the median inhibitory rate （IC50） of each group was calculated. The cell apoptosis rate after treatment by 17β-estradiol was detected by flow cytometry. ResultsThe expression intensities of ERβ mRNA in negative control, blank contro1 and siRNA group were 0.32±0.06, 0.28±0.05 and 0.05±0.02 respectively. The expression of ERβ mRNA was decreased significantly in siRNA group as compared with negative control group （P=0.032） and blank control group （P=0.015）. The expression intensities of ERβ protein in negative control, blank contro1 group and siRNA group were 0.18±0.04, 0.28±0.03 and 0.06±0.01, respectively. The expression of ERβ protein was decreased significantly in siRNA group as compared with negative control group （P=0.038） and blank control group（P=0.000）. After intervention with different concentrations of 17β-estradiol, the survival rate was decreased significantly in transfected group as compared with negative control group and blank control group ; The value of half maximal inhibitory concentration （IC50） in negative control , blank contro1 and siRNA group were （28.34±4.32）%, （27.66±3.04）% and （8.34±1.22）%, respectively. the growth of the lung cancer cell LTEP-a2 was significantly inhibited in siRNA group as compared with negative c

关 键 词：肺癌 性激素 RNA干扰 雌激素β基因 脱噬作用 

分 类 号：R734.2[医药卫生—肿瘤]