Lin28A过表达对胃癌细胞糖酵解的抑制作用及其机制  

Inhibitory effect of over-expression of Lin28A on glycosis in human gastric cancer cells and the underlying mechanism

作　　者：宋虎[1] 徐为[1] 宋军[1] 梁勇[1] 徐溢新[1] 许腾[1] 郑骏年 彭俊生[3] 

机构地区：[1]徐州医学院附属医院胃肠外科,江苏省徐州221002 [2]江苏省肿瘤生物治疗重点实验室 [3]中山大学附属第六医院食管胃肠外科,广州510655

出　　处：《中华临床营养杂志》2015年第4期234-237,共4页Chinese Journal of Clinical Nutrition

基　　金：江苏省高校自然科学研究面上项目（13KJB320029）;江苏省六大人才高峰项目（2014-WSW-032）

摘　　要：目的探讨Lin28A基因对胃癌细胞糖酵解的作用及其分子机制。方法利用慢病毒载体建立Lin28过表达胃癌BGC-823细胞株，以生物化学法检测细胞培养液乳酸含量，酶联免疫吸附试验（ELISA）法检测细胞内磷酸果糖（PFK）含量，蛋白质印迹法检测Lin28A、缺氧诱导因子1仅（HIF-1α）、葡萄糖转运蛋白1（GLUT-1）、丙酮酸激酶M2（PKM2）蛋白水平表达，反转录（RT）-PCR法检测let-7aRNA水平，膜联蛋白（Annexin）V-别藻蓝蛋白（APC）单染法检测细胞凋亡。结果Lin28A蛋白在转染组的表达水平明显高于对照组和空载组（均P〈0．001）。转染组let-7aRNA表达水平（0．602±0．017）明显低于空载组（1．001±0．063）（P=0．0052）。转染组HIF-1α、GLUT-1、PKM2蛋白的表达水平明显低于其他两组（均P〈0．001）。对照组、空载组、转染组细胞培养液乳酸含量分别为（1．71±0．13）、（1．53±0．11）、（1．24±0．04）mmol／L，转染组明显低于对照组和空载组（P=0．0170、0．0310）；细胞内PFK含量分别为（3．71±0．13）、（3．49±0．14）、（1．79±0，05）mmol／L，转染组明显低于对照组和空载组（P=0．0140、0．036O）；凋亡率分别为5．02％±0．14％、7．16％±0．21％、10．39％±0．37％，转染组凋亡率明显高于对照组和空载组（P=0．0005、0．0007）。结论Lin28A过表达可以抑制let-7a表达，并通过抑制细胞的糖酵解促进人胃癌细胞凋亡。Objective To explore the effects of over-expression of Lin28A on glycolysis of gastric cancer cells and the underlying mechanism. Methods Human gastric cancer BGC-823 cells in vitro were stably transfected with lentivirus-mediated over-expression of Lin28A. Lactic acid levels of cell culture medium were detected with biochemical method, phosphofructose kinase （PFK） levels in cells with ELISA, the expression levels of Lin28A, hypoxia-inducible factor （HIF） -1α, glucose transporter （GLUT） -1, and pyruvate kinase M2 （PKM2） were detected using Western blotting, and let-7a RNA using reverse transcription-PCR. Annexin V- allophycocyanin （APC） staining method was used to assess cell apoptosis. Results Compared with the normal control （CON） group and the negative control （NC） group, Lin28A protein level in the over-expression （OE） group was significantly higher （both P 〈 0. 001 ）. RNA expression level of let-7a was significantly lower in the OE group than in the NC group （0. 602 ±0. 017 vs. 1. 001 ±0. 063, P =0. 005 2）. The levels of the HIF-1α, GLUT- 1, PKM2 proteins were all significantly lower in the OE group than in the CON and the NC groups （ all P 〈0. 001 ）. The lactic acid levels of cell culture medium in the CON, the NC, and the OE groups were （ 1.71 ± 0. 13 ）, （ 1.53 ± 0. 11 ）, （ 1.24 ± 0. 04） mmol/L, which was significantly lower in the OE group than in the CON group and the NC group （P =0. 017 0, 0. 031 0）. Intracellular PFK level in the OE group was also significantly lower than in the CON group and the NC group [ （1.79 ±0.05） mmol/L vs. （3.71 ±0. 13） mmol/L, P = 0. 014 0 ; （ 1.79 ± 0. 05 ） mmol/L vs. （ 3.49 ± 0. 14） mmol/L, P = 0. 036 0 ]. Apoptosis rates of the CON, the NC, and the OE groups were 5.02% ±0. 14%, 7. 16% ±0. 21%, 10. 39% ±0. 37%, respectively, which was significantly higher in the OE group than in the CON group and the NC group （P = 0. 000 5, 0. 000 7）. Conclusion Over-expression of Lin28A can inh

关 键 词：Lin28A 胃癌 糖酵解 凋亡 

分 类 号：R735.2[医药卫生—肿瘤] R730.2[医药卫生—临床医学]