PHA-665752通过下调JAK2/STAT3通路调控PD-L1表达  

PHA-665752 regulates PD-L1 expression by downregulating the JAK2/STAT3 pathway

作　　者：邵倩倩 宋雪翼 涂丽 于滢 刘洋 丁晨冉 唐小龙 SHAO Qian-qian(Anhui University of Science&Technology,Huainan 232001,Anhui,China)

机构地区：[1]安徽理工大学医学院生物化学与分子生物学教研室,安徽淮南232001

出　　处：《牡丹江医学院学报》2024年第5期8-14,21,共8页Journal of Mudanjiang Medical University

基　　金：国家自然科学基金项目(82071862);安徽理工大学研究生创新基金项目(2023CX2152)。

摘　　要：目的探究索拉非尼(Sorafenib tosylate,SFB)耐药的肝细胞癌(Hepatocellular carcinoma,HCC)中,程序性细胞死亡蛋白配体1(PD-L1)的表达调控机制,关注细胞间质上皮转换因子(cellular-mesenchymal epithelial transition factor,c-Met)对PD-L1表达的影响,并寻找克服耐药性的潜在治疗策略。方法利用HepG2 ^(SR)细胞系作为研究对象,该细胞系对索拉非尼具有获得性耐药性,并表现出c-Met和PD-L1的高表达。并通过平板细胞克隆形成实验、Edu-594染色、Transwell小室、Matrigel基质胶、划痕愈合实验和CCK-8评估细胞增殖、迁移和侵袭能力。利用线粒体膜电位检测(JC-1试剂盒)膜电位变化,反映细胞凋亡情况。采用PHA-665752(c-Met的特异性抑制剂),观察其对c-Met磷酸化和PD-L1表达的影响。并通过蛋白免疫印迹研究PHA-665752对JAK2/STAT3信号通路激活的抑制效果,慢病毒介导的基因敲低技术来评估STAT3对PD-L1表达的影响。结果c-Met和PD-L1在HepG2 ^(SR)中高表达,PHA-665752作为c-Met抑制剂不仅能有效降低HepG2 ^(SR)细胞的增殖能力,迁移能力,提高细胞凋亡率,而且PHA-665752还抑制了JAK2/STAT3信号通路的激活。在敲低STAT3的细胞中,PD-L1的表达也相应下降。结论c-Met通过JAK2/STAT3信号通路在调控PD-L1的表达中起着重要作用。Objective To investigate the regulatory mechanism of programmed cell death protein ligand 1(PD-L1)expression in sorafenib tosylate(SFB)-resistant hepatocellular carcinoma(HCC)and to examine the impact of cellular-mesenchymal epithelial transition factor(c-Met)on PD-L1 expression,with the aim of identifying potential therapeutic strategies to overcome drug resistance.Methods The HepG2 ^(SR) cell line,which exhibits acquired resistance to sorafenib and high expression of c-Met and PD-L1,was used as the research model.Assessments of cell proliferation,migration,and invasion were conducted using a variety of assays,including the plate cell cloning formation assay,Edu-594 staining,Transwell assay,Matrigel matrix gel invasion assay,wound healing assay and CCK-8 assay.Changes in mitochondrial membrane potential,a marker of apoptotic activity,were detected using the JC-1 kit.PHA-665752,a specific inhibitor of c-Met,was employed to observe its effects on c-Met phosphorylation and PD-L1 expression.Western blot was employed to investigate the inhibitory effects of PHA-665752 on JAK2/STAT3 signaling pathway activation.Lentiviral-mediated gene knockdown technology was utilized to assess the impact of STAT3 on PD-L1 expression.Results High expression of c-Met and PD-L1 was observed in the HepG2 ^(SR) cells.Treatment with PHA-665752 led to a significant reduction in the proliferation and migration capabilities of HepG2 ^(SR) cells and an increase in the rate of apoptosis.Additionally,PHA-665752 inhibited the activation of the JAK2/STAT3 signaling pathway.In cells with STAT3 knockdown,a corresponding decrease in PD-L1 expression was observed.Conclusion c-Met plays a crucial role in the regulation of PD-L1 expression through the JAK2/STAT3 signaling pathway in sorafenib-resistant HCC.

关 键 词：C-MET PD-L1 JAK2-STAT3 肝细胞癌 索拉非尼 

分 类 号：R735.1[医药卫生—肿瘤]