中国癌症杂志 ›› 2018, Vol. 28 ›› Issue (2): 117-122.doi: 10.19401/j.cnki.1007-3639.2018.02.006

• 论著 • 上一篇    下一篇

XB130基因对肝细胞癌细胞增殖的影响及其机制

许文芳1,费迎明2,周建康2,陈将南1,周亚娣1,吕秋琼1   

  1. 1. 绍兴市立医院检验科,浙江 绍兴 312000 ;
    2. 绍兴市立医院消化内科,浙江 绍兴 312000
  • 出版日期:2018-02-28 发布日期:2018-03-08
  • 通信作者: 吕秋琼 E-mail:296882110@qq.com

The role of XB130 gene in proliferation of hepatocellular carcinoma cells and its mechanism

XU Wenfang1, FEI Yingming2, ZHOU Jiankang2, CHEN Jiangnan1, ZHOU Yadi1, LÜ Qiuqiong1   

  1. 1. Department of Clinical Laboratory, Shaoxing Municipal Hospital, Shaoxing 312000, Zhejiang Province, China; 2. Department of Gastroenterology, Shaoxing Municipal Hospital, Shaoxing 312000, Zhejiang Province, China
  • Online:2018-02-28 Published:2018-03-08
  • Contact: LÜ Qiuqiong E-mail: 296882110@qq.com

摘要: 背景与目的:XB130蛋白在肿瘤细胞增殖和侵袭中有重要作用,但在肝细胞癌(hepatocellular carcinoma,HCC)中的研究却极少,其作用至今仍不清楚。该研究拟探讨XB130基因对HCC细胞增殖能力的影响及其可能的下游机制。方法:采用蛋白[质]印迹法(Western blot)检测HCC细胞系Huh7、HepG2、SNU449及正常肝脏细胞系HL7702内XB130蛋白的表达。将XB130-siRNA转染Huh7细胞后,采用细胞计数试剂盒-8(cell counting kit-8,CCK-8)检测Huh7细胞活性,流式细胞术检测Huh7细胞周期,Western blot检测p-AKT、p-GSK3β、cyclin D1及p-Rb的蛋白表达水平,反转录聚合酶链反应(reverse transcription-polymerase chain reaction,RT-PCR)检测E2F/DP-1的靶基因cyclin E1、c-MycPCNA的mRNA表达水平。结果:XB130蛋白在Huh7、HepG2、SNU449和HL7702细胞中的相对表达量分别为0.66±0.10、0.78±0.11、0.83±0.08和0.32±0.06,各HCC细胞与HL7702细胞的差异均有统计学意义(P均<0.01)。XB130-siRNA转染可成功抑制Huh7细胞中XB130蛋白的表达。沉默XB130后,Huh7细胞活性在72 h后明显减弱(P<0.001),48 h后G0/G1期细胞比例升高,S和G2/M期细胞比例降低(P均<0.01),p-AKT、p-GSK3β、cyclin D1及p-Rb的蛋白表达下降(P均<0.01),cyclin E1、c-MycPCNA的mRNA表达下降(P 均<0.001)。结论:XB130通过对细胞周期蛋白及下游转录因子的调控,影响HCC细胞增殖。

关键词: 肝细胞癌, XB130, siRNA转染, 细胞增殖, 细胞周期

Abstract: Background and purpose: XB130 protein plays an important role in proliferation and invasiveness of tumor cells. However, there is little research on the role of XB130 protein in hepatocellular carcinoma (HCC) and the effect of XB130 is still unclear. This study investigated the role of XB130 gene in the proliferation of HCC cell and its potential mechanism. Methods: The protein expressions of XB130 in HCC cell lines, Huh7, HepG2 and SNU449, and liver cell line HL7702 were detected by Western blot. Huh7 cells were transfected with XB130-siRNA. Then cell viability was measured using cell counting kit-8 (CCK-8), and cell cycle was examined by flow cytometry. Protein expressions of p-AKT, p-GSK3β, cyclin D1 and p-Rb were detected by Western blot, while mRNA expression levels of E2F/DP1 target genes (cyclin E1, c-Myc and PCNA) were measured by reverse transcription-polymerase chain reaction (RT-PCR). Results: The relative protein expressions of XB130 in Huh7, HepG2, SNU449 and HL7702 cells were 0.66±0.10, 0.78±0.11, 0.83±0.08 and 0.32±0.06, respectively. The difference between HCC cell lines and HL7702 cell line was statistically significant (P<0.01). The transfection efficacy of XB130-siRNA was confirmed to be highly effective in Huh7 cells, and the viability of XB130-siRNA transfected Huh7 cells declined 72 h after transfection (P<0.001). The ratio of Huh7 cells in G0/G1 phase was increased, while the ratio in S or G2/M was decreased 48 h after XB130-siRNA transfection (P<0.01). In addition, compared with negative control, protein expressions of p-AKT, p-GSK3β, cyclin D1 and p-Rb, and mRNA expression levels of cyclin E1, c-Myc and PCNA were all decreased in XB130-siRNA transfected Huh7 cells (P<0.001). Conclusion: XB130 promotes the proliferation of HCC cells by regulating cell cycle-related proteins and downstream transcription factors.

Key words: Hepatocellular carcinoma, XB130, siRNA transfection, Proliferation, Cell cycle