GBP2和STAT1协同调控肝细胞癌的发展

GBP2 and STAT1 Synergistically Regulate the Development of HCC

目的研究肝细胞癌(HCC)组织中鸟苷酸结合蛋白2(GBP2)的表达及其对HCC的影响机制。方法选取南昌大学第二附属医院78例(男65例,女13例)HCC患者为研究对象,提取HCC肿瘤组织及癌旁正常组织;采购HCC细胞系(SK-HEP-1、HCCLM3和Huh-7)和正常肝细胞HL7702用于实验;选择4周龄雄性小鼠随机分为shGBP2干扰组(n=5)和shCtrl组(n=5)。分析GBP2在HCC中的表达水平及其与总生存期的关系;用短发夹RNA稳定敲低HCC细胞中的GBP2,并筛选相关性最强的GBP2共表达基因,通过多种体内、外细胞生物学实验研究其对肿瘤细胞的影响并阐明机制。结果HCC癌组织中GBP2表达水平显著高于癌旁正常组织(P<0.001),GBP2的表达水平与HCC的分期(P=0.001)和T浸润显著正相关(P=0.001),GBP2的高表达患者生存期低于低表达患者(P<0.001)。shGBP2干扰组小鼠(GBP2敲除)HCCLM3和SK-HEP 1细胞较shCtrl组小鼠增殖能力减弱(P<0.001)、迁移率显著较低(P<0.001)、凋亡力增强(P<0.01);shGBP2干扰组小鼠肿瘤生长较shCtrl组慢(P<0.001)且shGBP2组小鼠中的平均肿瘤重量显著轻于shCtrl组(P<0.001)。转录激活因子1(STAT1)和GBP2表达之间的正相关指数最高(r=0.248),HCC肿瘤细胞(HCCLM3、SK-HEP-1)中STAT1的表达高于正常肝细胞HL-7702(P<0.001)。GBP2过表达HCC细胞凋亡率最低(P<0.001),而STAT1敲除细胞凋亡率最高(P<0.05)。STAT1敲低的HCC细胞增殖能力显著降低(P<0.001)。将GBP2过表达的HCC细胞STAT1敲低后增殖力降低(P<0.001)、迁移力减弱(P<0.05)、凋亡率增加(P<0.001)。结论GBP2下调可抑制肝癌细胞的增殖、迁移和致瘤能力,GBP2敲低的HCC细胞表现出明显的凋亡增强。HCC患者中GBP2与STAT1的表达呈正相关,STAT1的敲低可部分缓解GBP2过表达对HCC细胞的驱动作用。GBP2和STAT1协同调控HCC的发生发展,可能成为HCC治疗的靶点。

Objective To investigate the expression of guanylate binding protein 2(GBP2)in hepatocellular carcinoma(HCC)and its effects on HCC.Methods A total of 78 HCC patients(65 males and 13 females)from the Second Affiliated Hospital of Nanchang University were selected for the study,and HCC tumor tissues and paracancerous tissues were extracted.HCC cell lines(SK-HEP-1,HCCLM3,and Huh-7)and normal hepatocytes HL7702 were procured for experiment.Four-week-old male mice were randomly divided into the shGBP2 interference group(n=5)and the shCtrl group(n=5).The expression level of GBP2 in HCC and its relationship with overall survival were analyzed.Short hairpin RNA was employed to achieve stable knockdown of GBP2 in HCC cells,and the most relevant co-expression genes of GBP2 were screened.Their effects on tumor cells were investigated through a series of in vivo and in vitro cell biology experiments.Results The expression level of GBP2 in HCC tissues was significantly higher than that in paracancerous tissues(P<0.001);GBP2 expression level was positively correlated with HCC stage(P=0.001)and T infiltration(P=0.001);patients with high GBP2 expression had lower survival rates compared to those with low expression(P<0.001).In the shGBP2 interference group(GBP2 knockout),HCCLM3 and SK-HEP 1 cells exhibited decreased proliferation(P<0.001),significantly lower mobility(P<0.001),and increased apoptosis(P<0.01).Tumor growth in the shGBP2 interference group was slower than that in the shCtrl group(P<0.001),and the mean tumor weight in the shGBP2 interference group was significantly lower than that in the shCtrl group(P<0.001).The positive correlation index between the expression of STAT1 and GBP2 was the highest(r=0.248).The expression of STAT1 in HCC tumor cells(HCCLM3,SK-HEP-1)was significantly higher than that in normal hepatocytes HL-7702(P<0.001).GBP2-overexpressed HCC cells exhibited the lowest apoptosis rate(P<0.001),while STAT1 knockout cells showed the highest apoptosis rate(P<0.05).The proliferation ability of HCC cells with low STAT1 knockdown was markedly decreased(P<0.001).Furthermore,both proliferation and migration of GBP2-overexpressed HCC cells decreased significantly after STAT1 knockdown(P<0.05),accompanied by an increase in apoptosis rate(P<0.001).Conclusion GBP2 downregulation can inhibit the proliferation,migration,and tumorigenicity of HCC cells,and GBP2 knockdown in HCC cells shows a marked enhancement of apoptosis.The expression of GBP2 is positively correlated with that of STAT1 in HCC patients,and STAT1 knockdown can partially alleviate the driving effect of GBP2 overexpression on HCC cells.GBP2 and STAT1 synergistically regulate the development and progression of HCC,which may become a target for HCC treatment.