摘要
目的观察Girdin基因在肝癌细胞表达及抑制其表达对癌细胞增殖、凋亡的影响。方法Western blot检测人正常肝细胞HL-7702及肝癌Huh7、HepG2、MHCC97H、HCCLM3细胞Girdin的蛋白表达；Girdin的小干扰RNA（siRNA）通过Lipofectamine？2000转染HCCLM3细胞，并设置阴性对照组和空白对照组，收集转染48 h的细胞，Western blot检测Girdin、细胞核增殖抗原（Ki-67）、半胱氨酰天冬氨酸特异性蛋白酶（Caspase）家族含Caspase-3、B细胞淋巴瘤/白血病-2（bcl-2）家族促凋亡基因bcl-2相关X蛋白（bax）及酪氨酸激酶2（JAK2）/信号转导与转录激活因子3（STAT3）通路磷酸化的酪氨酸激酶2（p-JAK2）、磷酸化的信号转导与转录激活因子3（p-STAT3）的蛋白表达；细胞计数试剂盒（CCK-8）法检测转染的24、48、72 h各组细胞的活力；流式细胞术检测转染48 h的细胞凋亡率。结果Girdin在HepG2（0.355±0.043）、HCCLM3（0.599±0.068）、MHCC97H（0.462±0.055）、Huh7（0.378±0.049）肝癌细胞中的表达均显著高于在正常肝细胞HL-7702（0.112±0.013）的表达（PHepG2＝0.001；PHCCLM3＝0.000；PMHCC97H＝0.000；PHuh7＝0.001）；Girdin-siRNA组在24 h（0.247±0.032）、48 h（0.477±0.043）和72 h（0.666±0.073）的细胞活力均显著低于空白对照组（0.385±0.045）、（0.711±0.068）、（0.936±0.096），在48 h的细胞凋亡率（16.18±1.38）%显著高于空白对照组（1.87±0.39）%，Ki-67（0.162±0.019）、p-JAK2（0.096±0.011）、p-STAT3（0.057±0.009）蛋白表达显著低于空白对照组（0.626±0.073）、（0.170±0.021）、（0.145±0.018），Caspase-3（0.257±0.032）和bax（0.279±0.032）的蛋白表达显著高于空白对照组（0.120±0.013、0.135±0.015）（PKi-67＝0.000；Pp-JAK2＝0.006；Pp-STAT3＝0.002；PCaspase-3＝0.002；Pbax＝0.002）。结论Girdin基因在肝癌细胞高表达，通过RNA干扰抑制其表达通过下调JAK2/STAT3信号降低癌细胞的增殖及诱�
ObjectiveTo investigate the Expression of Girdin gene in hepatocellular carcinoma cells and inhibition of its expression on proliferation and apoptosis of cells.MethodsThe expression of Girdin protein in HL-7702 normal liver cells and liver cancer Huh7, HepG2, MHCC97H, HCCLM3 cells were detected by Western blotting; Girdin small interfering RNA （siRNA） were transfected into HCCLM3 cells by Lipofectamine？2000, and set the negative control group and blank control group, the expression of proliferation cell nuclear antigen （Ki-67）, cysteinyl aspartate-specific protease （Caspase）-3, B cell lymphoma/leukemia-2 associated X protein （bax）, phosphorylation of janus kinase 2 （JAK2） and signal transducer and activators of transcription 3 （STAT3） protein were detected by Western blotting; cells activity were detected by cell counting kit-8 （CCK-8） assay in transfected 24, 48 and 72 h; cell apoptosis were detected by flow cytometry in transfected 48 h.ResultsThe expression of Girdin in HepG2 （0.355±0.043）, HCCLM3 （0.599±0.068）, MHCC97H （0.462±0.055）, Huh7 （0.378±0.049） HCC cells was significantly higher than that in normal liver cells HL-7702 （0.112±0.013, PHepG2=0.001; PHCCLM3=0.000; PMHCC97H=0.000; PHuh7=0.001）; cell activity in Girdin-siRNA group in 24 h （0.247±0.032）, 48 h （0.477±0.043） and 72 h （0.666±0.073） were significantly lower than the control group （0.385±0.045）, （0.711±0.068）, （0.936±0.096）, the apoptosis rate （16.18±1.38）% in 48 h was significantly higher than the blank control group （1.87±0.39）%, the expression of Ki-67 （0.162±0.019）, p-JAK2 （0.096±0.011） and p-STAT3 （0.057±0.009） protein were significantly lower than the control group （0.626±0.073）, （0.170±0.021）, （0.145±0.018）, the expression of Caspase-3 （0.257±0.032） and bax （0.279±0.032） protein were significantly higher than the control group （0.120±0.013, 0.135±0.015） （PKi-67=0.000; Pp-JAK2=0.006; Pp-STAT3=0.002;
出处
《中华实验外科杂志》
CSCD
北大核心
2017年第12期2109-2112,共4页
Chinese Journal of Experimental Surgery
基金
河南省教育厅科技研究重点项目（13A32041）
关键词
Girdin基因
肝癌
增殖
凋亡
酪氨酸激酶2/信号转导与转录激活因子3信号通路
Girdin gene
Hepatocellular carcinoma
Proliferation
Apoptosis
Janus kinase 2/signal transducer and activators of transcription 3 signaling pathway
