siRNA靶向抑制SPAG_9对HepG_2细胞增殖、凋亡及JNK信号通路的影响  

Effects of Small Interfering RNA(siRNA)-Mediated Knockdown of SPAG_9 on HepG_2 Proliferation,Apoptosis and JNK Signal Pathway

作　　者：罗书笛 任碧琼[2] 刘俊龙 陈维 Luo Shudi;Ren Biqiong;Liu Junlong(Clinical Medical College of Hunan University of Chinese Medicine,Hunan 410007,China)

机构地区：[1]湖南中医药大学临床医学院,长沙410007 [2]湖南省第二人民医院,长沙410007

出　　处：《医学研究杂志》2019年第1期103-107,共5页Journal of Medical Research

基　　金：湖南省研究生创新基金资助项目(CX2016B381)

摘　　要：目的探讨精子相关抗原9(SPAG9)在肝癌细胞HepG_2中表达对HepG_2增殖分化及凋亡的影响,并初步揭示SPAG9与JNK信号通路的关系。方法采用细胞免疫荧光检测SPAG9在HepG_2中的表达及定位,siRNA技术敲低SPAG9的表达,Western blot法检测转染后SPAG9及JNK蛋白的表达,CCK8(cell counting kit-8)实验检测肿瘤细胞的增殖能力,流式细胞术检测肿瘤细胞的周期及凋亡。结果 SPAG9在HepG_2中表达,定位于细胞质和细胞核。与对照组细胞比较,转染SPAG9siRNA使JNK蛋白的表达下调,差异有统计学意义(P<0.05)。与对照组比较,转染SPAG9si RNA的HepG_2增殖能力显著降低(P<0.01),细胞凋亡显著增加(P<0.01),细胞周期阻滞于S期。结论 HepG_2细胞表达SPAG9蛋白,定位于细胞质和细胞核;SPAG9促进HepG_2细胞增殖分化、抑制凋亡与JNK信号通路有关;敲低SPAG9的表达使HepG_2细胞增殖阻滞于S期。Objective To explore the expression of SPAG 9 in hepatoma HepG 2 cell line and its effects on cell proliferation,differentiation and apoptosis,and reveal the relationship between SPAG 9 and JNK signal pathway.Methods The expression and location of SPAG 9 in HepG 2 was detected by cell immunofluorescence.SiRNA transfection was used to knockdown the expression of SPAG 9.Western blot was used to detect the expression of SPAG 9 and JNK protein after siRNA transfection.Cell Counting Kit-8 assay was used to detect the effect of SPAG 9 on the proliferation of HepG 2 after siRNA transfection.Flow cytometry was used to detect cell cycle and apoptosis of HepG 2 after transfection.Results SPAG 9 was positively expressed in HepG 2 cell line,and located on the cytoplasm and nucleus of HepG 2 cells.Compared with control group,the expression of JNK protein was significantly decreased after SPAG 9 siRNA transfection(P<0.05).Compared with control group,SPAG 9 knockdown significantly reduced HepG 2 cell proliferation(P<0.01)and increased cell apoptosis(P<0.01).SPAG 9 knockdown also resulted in cell cycle arrest in S phase.Conclusion SPAG 9 is positively expressed in HepG 2 cells,and located on cytoplasm and nucleus.SPAG 9 promotes cell proliferation and differentiation,inhibits cell apoptosis in liver cancer possibly through JNK signal pathway.SPAG 9 knockdown could arrest cell cycle at S phase.

关 键 词：精子相关抗原9 HEPG2 JNK 增殖 凋亡 

分 类 号：R73[医药卫生—肿瘤]