PKM2对人胃癌细胞HMGB1释放的影响

目的:探讨M2型丙酮酸激酶(pyruvate kinase M2,PKM2)对胃癌细胞高迁移率族蛋白B-1(high mobility group box 1,HMGB1)释放的作用。方法:通过siRNA干扰抑制人胃癌BGC823细胞PKM2的表达，Western blot检测各组细胞PKM2蛋白的表达，以评价PKM2 siRNA的转染效率；CCK-8法检测各组细胞活力，分析干扰PKM2的表达对胃癌BGC823细胞增殖的影响；Western blot检测各组细胞HMGB1的表达以及ELISA检测各组细胞培养液HMGB1的水平，分析PKM2对胃癌BGC823细胞HMGB1的作用。结果:与空载体pU6组相比，PKM2 siRNA 组PKM2的表达显著降低(P＜0.01)，表明siRNA有效地抑制胃癌BGC823细胞PKM2的表达；CCK-8结果显示，PKM2 siRNA 组细胞活力明显低于空载体pU6组(P＜0.001)，表明干扰PKM2抑制胃癌BGC823细胞增殖；此外，PKM2 siRNA 组HMGB1的表达以及细胞外HMGB1的水平显著降低(P＜0.01)，表明干扰PKM2抑制人胃癌BGC823细胞HMGB1的释放。结论:PKM2促进人胃癌BGC823细胞HMGB1的释放。

The effect of PKM2 on the release of HMGB1 in human gastric cancer BGC823 cell

Objective:To explore the effect of pyruvate kinase M2 (PKM2) on the release of high mobility group box 1 (HMGB1) in human gastric cancer cell.Methods:siRNA was used to reference the expression of PKM2 in gastric cancer BGC823 cell.The effect of inferenced PKM2 on the invasion of gastric cancer BGC823 cell was analysed,and the cell viability was detected by Cell Counting Kit (CCK-8).In order to evaluate the role of PKM2 in the HMGB1 release of gastric cancer BGC823 cell,the Western blot and ELISA were applied to measure the level of HMGB1.Results:Compared to pU6 group,the expression of PKM2 in PKM2 siRNA group was dramatically decreased (P＜0.01),which indicated that the siRNA plasmid was successfully transfected into gastric cancer BGC823 cell.The result from CCK-8 showed that the cell viability in PKM2 siRNA group was declined compared to pU6 group (P＜0.001),suggesting inhibitory role of PKM2 siRNA in the gastric cancer BGC823 cell invasion.More importantly,PKM2 siRNA decreased level of HMGB1 in gastric cancer BGC823 cell (P＜0.01).Conclusion:Reference PKM2 by siRNA can inhibit the HMGB1 release of human gastric cancer BGC823 cell.