RNA干扰NGAL基因表达对乳腺癌细胞增殖凋亡的影响机制研究  

Study on the mechanism of effect of RNA interference NGAL gene expression on proliferation and apoptosis of breast cancer cells

作　　者：周庆[1] 胡清林[1] 郭智慧[1] 李晓诗 ZHOU Qing;HU Qing-Lin;GUO Zhi-Hui;et al.(Department of Thyroid and Breast Surgery, the First Affiliated Hospital of Chengdu Medical College, Chengdu, Sichuan 610500, China)

机构地区：[1]成都医学院第一附属医院甲乳外科,四川成都610500

出　　处：《中国妇幼保健》2018年第13期3056-3060,共5页Maternal and Child Health Care of China

摘　　要：目的分析RNA干扰中性粒细胞相关载脂蛋白(NGAL)基因的表达对乳腺癌细胞增殖凋亡的影响及机制。方法利用蛋白质印迹(Western Blot)方法检测乳腺癌组织及人乳腺癌MCF-7、T47D和MDA-MB-231细胞株中NGAL的蛋白表达;小干扰RNA(siRNA)-NC及NGAL-siRNA转染人乳腺癌MCF-7细胞,未转染任何siRNA作为对照组,检测转染48 h后各组细胞中NGAL的蛋白表达情况;利用细胞计数试剂盒(CCK8)方法检测细胞的增殖情况;利用流式细胞仪检测细胞的凋亡;利用Western Blot方法检测活化的含半胱氨酸的天冬氨酸蛋白水解酶3(Cleaved Caspase-3)、磷脂酰肌醇-3(PI3K)、丝氨酸苏氨酸激酶(AKT)及磷酸化的丝氨酸苏氨酸激酶(p-AKT)的蛋白表达情况。结果乳腺癌组织中NGAL的蛋白表达显著高于癌旁组织(P<0.05),NGAL于乳腺癌MCF-7细胞中的蛋白表达最高,选择其作为后续研究;siRNA-NC组NGAL、Cleaved Caspase-3、PI3K及p-AKT蛋白表达和细胞存活率及凋亡率与相应的对照组比较差异无统计学意义(P>0.05),而NGAL-siRNA组NGAL、PI3K和p-AKT蛋白表达及细胞存活率均显著低于相应的对照组(P<0.05),NGAL-siRNA组的细胞凋亡率及Cleaved Caspase-3蛋白表达均显著高于相应的对照组(P<0.05)。结论 NGAL基因在乳腺癌组织及细胞中高表达,抑制NGAL基因的表达会显著降低细胞增殖及促进细胞凋亡,其机制与PI3K/AKT信号通路的调控有关。Objective To analyze the effect and mechanism of RNA interference neutrophil gelatinase-associated lipocalin （NGAL） gene expression on proliferation and apoptosis of breast cancer cells. Methods Western Blot was used to detect NGAL protein expression in breast cancer tissues and human breast cancer MCF-7, T47D, and MDA-MB-231 cells, siRNA-NC and NGAL-siRNA were used to trans- feet human breast cancer MCF-7 cells, the cells not transfected by any siRNA were collected as control group. After 48 hours of transfection, the expression levels of NGAL protein in different groups were detected. CCK8 method was used to detect cell proliferation. Flow cy- tometry was used to detect cell apoptosis. Western Blot was used to detect the expression levels of Cleaved Caspase-3, PI3K, AKT, and p-AKT. Results The expression level of NGAL in breast cancer tissue was statistically significantly higher than that in paraeaneerous tissue （ P〈0. 05） . The expression level of NGAL in breast cancer MCF-7 cells was the highest, so it was selected for the tollow-up study. There was no statistically significant difference in the expression levels of NGAL, Cleaved Caspase-3, PI3K, and p-AKT, cell survival rates, and cell apoptotic rates between siRNA-NC group and control group （P〉0. 05） . The expression levels of NGAL, PI3K, and p-AKT, cell sur- vival rate in NGAL-siRNA group were statistically significantly lower than those in control group （ P〈0. 05 ） . Cell apoptotic rate and expression level of Cleaved Caspase-3 in NGAL-siRNA group were statistically significantly higher than those in control group （P〈0.05） . Con- clusion The expression of NGAL gene is high in breast cancer tissues and cells, the inhibition of NGAL gene expression can significantly reduce cell proliferation and promote apoptosis, and the mechanism is related to the regulation of PI3K/AKT signaling pathway.

关 键 词：RNA干扰 NGAL基因 乳腺癌 增殖 凋亡 

分 类 号：R737.9[医药卫生—肿瘤]