山东医药2019年第59卷第17期胃癌细胞中乳酸脱氢酶A表达变化 及其对EMT的影响陈妍\2，陈加华3,李小琴1(1江苏大学附属医院，江苏镇江212031 ；2徐州医科大学附属淮安医院;3沐阳仁慈医院)摘要：目的 观察胃癌细胞中乳酸脱氢酶A(LDH-A)的表达变化，并探讨沉默LDH-A表达对胃癌细胞上皮间 质转化(EMT)的影响。方法体外培养胃癌细胞株OCUM-1和正常胃黏膜上皮细胞株RGM-1,采用qRT-PCR法
检测细胞中LDH-A mRNA相对表达量,Western blotting法检测细胞中LDH-A蛋白表达情况。将OCUM-1细胞随机 分为3组,LDH-A siRNA组采用Lipfoctamine2000脂质体转染试剂对细胞转染LDH-A siRNA序列，转染对照组转染
siRNA无义对照序列，空白对照组只加入培养基。转染48 h后收集各组细胞，显微镜下观察胃癌细胞的形态学变
化，采用Western blotting法检测细胞中上皮细胞特征蛋白上皮性钙黏蛋白(E-cadherin)、细胞角蛋白(Cytokeratin) 和间质细胞特征蛋白神经性钙黏蛋白(N-cadherin)、锌指转录因子(Snail)、-连环蛋白(|3-catenin)、波形蛋白(Vim-
entin)的表达。结果 OCUM-1细胞中LDH-A mRNA及蛋白表达高于RGM-1细胞(P均＜0. 05)。LDH-A siRNA
组LDH-A mRNA及蛋白表达低于转染对照组和空白对照组(P均＜0. 05)。显微镜下可见LDH-A siRNA组细胞排 列紧密，细胞黏附性增加，表现为上皮细胞特性。LDH-A siRNA组E-cadherin、Cytokeratin蛋白表达水平高于转染对
照组和空白对照组，N-cadherin、Snail、［3-catenin、Vimentin蛋白表达水平低于转染对照组和空白对照组(P均＜
0.05)o结论 胃癌细胞中LDH-A表达升高;沉默LDH-A表达可上调胃癌细胞上皮细胞特征蛋白表达、下调间质
细胞特征蛋白表达,从而抑制胃癌细胞的EMT进程。关键词：胃癌细胞；胃黏膜上皮细胞;乳酸脱氢酶A；波形蛋白；上皮性钙黏蛋白；细胞角蛋白；神经性钙黏蛋 白；锌指转录因子；-连环蛋白；上皮间质转化doi ： 10. 3969/j. issn. 1002-266X. 2019.17. 006中图分类号:R735.2 文献标志码:A 文章编号1002-266X(2019) 174020-04：Expression changes of lactate dehydrogenase A in gastric cancer cells and its effect on
epithelial-mesenchymal transformationCHEN Yan , CHEN Jiahua, LI Xiaoqin(1 The Affiliated Hospital of Jiangsu University, Zhenjiang 212031 , China /： , ) and to explore the effect of silencing LDH-A expression on epithelial mesenchymal transformation (EMT) in gastric cancer
AbstractObjective To observe the expression changes of lactate dehydrogenase A LDH-Ain gastric cancer cells
(cells. Methods The gastric cancer cell line OCUM-1 and normal gastric mucosal epithelial cell line RGM-1 were cultured in vitrothe relative expression of LDH-A mRNA in cells was detected by qRT-PCRthe expression of LDH-A protein in
cells was detected by Western blotting. OCUM- 1 cells were randomly divided into 3 groupsLDH-A siRNA group was
, , transfected with LDH-A siRNA sequence using Lipfoctamine2000 liposome transfection reagentthe transfection control
, were observed under microscope, the expression of E-cadherin, Cytokeratin, N-cadherin, Snail, p-catenin, and Vimentin
um. After 48-hour transfectionthe cells of each group were collected and the morphological changes of gastric cancer cells
group was transfected with LDH-A siRNANC sequenceand the blank control group was only added with the culture medi-
, , was detected by Western blotting. Results The mRNA and protein expression of LDH-A in the OCUM-1 cells was higher than that in RGM-1 cells both P ＜0. 05 . The mRNA and protein expression of LDH-A in the LDH-A siRNA group was
lower than that in the transfection control group and blank control group all P ＜0. 05 . Under microscopythe cells in the LDH-A siRNA group were closely arrangedand the cell adhesion increasedshowing the characteristics of epithelial cells.
( ), ( , ), The protein expression levels of E-cadherin and Cytokeratin in the LDH-A siRNA group were higher than those in the trans-第一作者简介:陈妍（1986-），女，主治医师,主要研究方向为胃癌的基础与临床治疗。E-mail： chenyan86192@163.com 通信作者简介:李小琴（1967-），女，主任医师，主要研究方向为消化系统肿瘤。E-mail： lixiaoqin6699@126.com
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胃癌细胞中乳酸脱氢酶A表达变化及其对EMT的影响 - 论文
