siRNA抑制S100A4蛋白对人胃腺癌细胞增殖和凋亡及化疗敏感性的影响  

Effect of small interference RNA silenced S100A4 protein in hu- man gastric carcinoma cells on proliferation, apoptosis and chemotherapy sensitivity in vitro

作　　者：李鹏[1] 刘江伟[2] 韩振魁[1] 

机构地区：[1]新疆维吾尔自治区人民医院胃肠外科,乌鲁木齐830000 [2]兰州军区乌鲁木齐总医院动物实验科,乌鲁木齐830000

出　　处：《国际外科学杂志》2016年第5期300-304,F0003,共6页International Journal of Surgery

基　　金：基金项目：新疆维吾尔自治区人民医院院内项目（20120139）;中国博士后科学基金第48批面上项目（20100481517）

摘　　要：目的研究应用小干扰RNA沉默S100A4蛋白后对人胃腺癌细胞BGC-823增殖、凋亡及化疗敏感性的影响。方法人胃腺癌BGC-823细胞系转染siRNA，RT—PCR检测转染效果后将对数期胃腺癌细胞分为干扰组、阴性对照组、正常对照组，MTT检测不同浓度奥沙利铂对胃腺癌细胞中效浓度IC50值；绘制细胞生长曲线；TUNEL法检测细胞凋亡；RT—PCR检测各组细胞mRNA变化；Westernblot检测蛋白水平的变化，计量资料选用t检验，SPSS17．0分析RT—PCR检测各组细胞mRNA的变化，Westernblot检测蛋白水平的变化，细胞生长曲线描述细胞增殖，TUNEL法检测细胞凋亡，MTT法检测不同浓度奥沙利铂对胃腺癌细胞中效浓度IC50值。计量资料以（s±s）表示，多个样本比较采用单因素方差分析和LSD检验方法。P〈0．05为差异有统计学意义。结果RT—PCR结果显示BGC-823细胞转染S100A4siRNA48h后，S100A4mRNA的表达量分别为：（0．674±0．011）、（0．652±0．021）、（0．345±0．040），正常对照组与干扰组相比差异有统计学意义（P=0．012，P〈0．05），阴性对照组与干扰组差异均有统计学意义（P=0．000，P〈0．05），正常对照组与阴性对照组差异无统计学意义（P=0．380，P〉0．05）；Westernblot结果显示BGC-823细胞转染S100A448h后表达量明显下调分别为：（0．654±0．025）、（0．642±0．014）、（0．317±0．061），正常对照组与干扰组相比差异有统计学意义（P=0．01，P〈0．05），阴性对照组与干扰组差异均有统计学意义（P=0．000，P〈0．05），正常对照组与阴性对照组无统计学差异（P=0．341，P〉0．05）。S100A4siRNA转染后人胃腺癌BGC-823细胞增殖下降；TUNEL法结果显示转染后凋亡明显增加；MTT结果表明人胃腺癌BGC-823单用奥沙利铂中效浓度IC50分别为56．31μmol／L，转染后奥沙利铂中效浓度IC50为0．654μmol／L。结论本研究结果表明，Objective To study the application of small interfering RNA silencing S100A4 protein in hu- man gastric cancer cell BGC-823 proliferation, apoptosis and the effect of chemotherapy sensitivity. Methods Human gastric carcinoma cell line BGC -823 transfection siRNA, RT-PCR detected the changes of mRNA after transfection. Groups divided into interference group, negative control group and normal con- trol group. MTT test determined different concentrations of oxaliplatin in gastric cancer cells and calculated ICs0, then draw cell growth curve, TUNEL method to detect apoptosis, RT-PCR tested each cell mRNA changed, Western blot detected the change of the S100A4 protein. All data analysis by SPSS17.0, t test ap- plied, RT-PCR and Western blot results analysis by SPSS17.0, comparing multiple samples by using single factor analysis of variance and LSD test. P 〈 0.05 was statistically significant. Results RT-PCR results showed that BGC-823 cell transfection, S100A4mRNA expression quantity respectively after 48 hours： （0.674±0. 011）, （0. 652±0.021 ）, （0. 345 ±0. 040）, the interference group and normal control group were statistically significant （P = 0. 012, P 〈 0.05 ） and the negative control group with interference group differences were statistically significant （P =0. 000, P 〈0.05）, and normal control group was no statistical- ly significant difference with the negative control group （ P = 0. 380, P 〉 0. 380 ） ; Western blot results showed BGC -823 cell transfeetion S100A4 expression significantly lowered respectively after 48 hours ,there were （0. 654 ± 0.025 ）, （0. 642 ± 0. 014）, （0.317 - 0. 061 ）, the interference group and normal control group was statistically significant （ P = 0. 01, P 〈 0.05 ） , between negative control group and interference group were statistically significant （ P = 0. 000, P 〈 0.05 ）, normal control group and the negative control group had no significant difference （ P = 0. 341, P 〉 0. 341 ）. After S100A4- siRNA transfeeti

关 键 词：胃腺癌 小干扰RNA S100A4 细胞增殖 细胞凋亡 化疗敏感性 

分 类 号：R735.2[医药卫生—肿瘤]