摘要
目的:研究糖原合成酶激酶-3β(glycogen synthase kinase,GSK-3β)对子宫内膜样腺癌已分化细胞株Ishikawa、HEC-1-A及未分化细胞株KLE增殖和侵袭能力的影响。方法:用GSK-3β小分子干扰RNA(GSK-3βsiRNA)转染上述3种细胞株,采用Western印迹法检测GSK-3β蛋白及凋亡相关蛋白caspase-3的表达;Brdu掺入实验检测细胞增殖率;FCM法检测细胞周期及凋亡率;Transwell侵袭试验检测对细胞侵袭、运动能力的影响;明胶酶谱法检测细胞分泌基质金属蛋白酶-2(MMP-2)的情况。结果:转染GSK-3βsiRNA后,3种细胞株中GSK-3β蛋白的表达均低于对照组(P<0.05);Ishikawa、HEC-1-A细胞Brdu掺入率降低,S期细胞数比例减少,凋亡率增加,caspase-3表达上调;细胞侵袭能力降低,与对照组相比差异有统计学意义(P<0.01)。但是KLE细胞株与对照组相比,增殖和侵袭能力均无明显差异(均为P>0.05)。结论:GSK-3β能促进已分化的子宫内膜样腺癌细胞株Ishikawa、HEC-1-A的增殖和侵袭;但是对未分化的KLE细胞株的增殖和侵袭能力无明显影响。
Objective：To investigate the effects of glycogen synthase kinase-3β（ GSK-3β） on the proliferation and invasion abilities of the three cell lines with endometrioid adenocarcinoma Ishikawa, HEC-1A （differentiated） and KLE （undifferentiated）. Methods：Three human endometriold adenocarcinoma cell lines Ishikawa, HEC-1A, and KLE were transfected with GSK-3β siRNA. The expression of GSK-3β protein and apoptosis-related protein caspase-3 were examined by Western blotting. The proliferation of cells transfected with GSK-3β siRNA or negative control siRNA were meausred with BrdU incorporation assay while cell cycle distribution and apoptotic ratio were detected by flow cytometry （ FCM ）. The invasion and migration of cells were tested by Matrigel invasion assay. MMP-2 secretion was assessed by gelatin zymography. Results：The GSK-3β siRNA had high inhibitory effect on the expression of GSK- 3β gene. The level of GSK-3β protein was significantly lower in the three cell lines compared with control （P 〈0.05）. Compared with cells transfected with negative control siRNA or non transfected cells, the BrdU incoporation ratio was significantly decreased in Ishikawa and HEC-1-A cells after transfected with GSK-3β siRNA （ P 〈 0.05 ）. The proportion of cells in S phase was reduced and the apoptotic rate was increased while the expression of caspase 3 protein was up-regulated and cell invasion ability was down-regulated （P 〈0.01 ）. But there was no statistical difference in the proliferation and invasion abilities between KLE cells and control （P 〉 0.05 ）. Conclusion： GSK-3β could promote the proliferation and invasion of the differentiated endometrioid adenocarcinoma cell lines Ishikawa and HEC-1-A, but had no effect on the undifferentiated cell line KLE.
出处
《肿瘤》
CAS
CSCD
北大核心
2008年第4期288-292,共5页
Tumor
基金
上海市医学重点学科建设资助项目(编号:05Ⅲ016)
