摘要
目的:探讨GRP94基因对肝癌细胞的影响。方法:用脂质体法转染GRP94 siRNA序列及阴性对照序列,采用实时荧光定量PCR及Western blotting方法检测转染后GRP94的mRNA和蛋白表达水平。以CCK-8和流式细胞仪来检测细胞增殖和细胞凋亡的情况。以划痕实验和Transwell方法检测转染后细胞的迁移和侵袭能力。结果:在肝癌细胞Hep G2中瞬时转染siRNA-GRP94序列及阴性对照序列后,其mRNA和蛋白表达显著减低;转染48 h后,细胞增殖能力明显下降,而细胞凋亡率明显上升。划痕实验检测发现24 h细胞愈合率明显抑制,Transwell实验发现迁移和侵袭至下室的细胞数目显著减少。结论:GRP94基因对肝癌细胞Hep G2凋亡有抑制作用,促进增殖、迁移和侵袭能力,为进一步研究GRP94在肝细胞肝癌发生和进展中的作用奠定了基础。
Objective： To investigate the effect of GRP94 gene on hepatocellular carcinomas cells.Methods： The siRNA sequence and negative control sequence of GRP94 were transfected to cells using lipidosome.The mRNA and protein expression levels of GRP94 in HepG2 cells were detected using qRT-PCR and Western blotting,respectively.The proliferation and apoptosis of cells were detected using CCK-8 and flow cytometry,respectively.The migration and invasion abilities of HepG2 cells were tested using wound healing and Transwell assay,respectively.Results： The mRNA and protein expression levels of GRP94 in HepG2 cells after transfecting siRNA-GRP94 sequence and NC sequence significantly decreased.After 48 h of transfection,the proliferation ability of cells significantly decreased,and the apoptosis rate of cells significantly increased.The scratch test showed that the 24 h healing rate of cells was significantly inhibited,and the Transwell assay showed that the number of migration and invasion cells significantly decreased.Conclusions： GRP94 gene can inhibit the apoptosis,and promote the proliferation,migration and invasion of HepG2 cells,which can provide an experiment basis in investigating the effects of GRP94 gene on the occurrence and development of hepatocellular carcinoma.
作者
和小华
林兰
顾健美
张小霞
王秋红
HE Xiao-hua;LIN Lan;GU Jian-mei;ZHANG Xiao-xia;WANG Qiu-hong(Department of Clinical Laboratory,The Affiliated Cancer Hospital of Nantong University,Nantong Jiangsu 226000;Department of Clinical Laboratory,Nantong Maternal and Child Health Care Hospital Affiliated to Nantong University,Nantong Jiangsu 226000,China)
出处
《蚌埠医学院学报》
CAS
2018年第9期1129-1133,共5页
Journal of Bengbu Medical College
基金
江苏省南通市卫生局青年基金(WQ2015057)
江苏省南通市科技局科技计划项目指导性课题(YYZ17024)
关键词
肝癌细胞
葡萄糖调节蛋白
增殖
凋亡
迁移
侵袭
hepatoma cells
glucose-regulated 94
proliferation
apoptosis
migration
invasion
