TBC1D5通过JAK/STAT通路对肝细胞癌进展的影响  

Effect of TBC1D5 on hepatocellular carcinoma progression via JAK/STAT pathway

作　　者：韦豪伟 陶学文 余德才[1] Wei Haowei;Tao Xuewen;Yu Decai(Dept of Hepatobiliary and Transplantation Sugery,Affiliated Drum Tower Hospital,Medical School of Nanjing University,Nanjing 210008;Dept of Hepatobiliary,Pancreatic and Fransplantation Surgery,The First Affiliated Hospital of Anhui Medical University,Hefei 230022)

机构地区：[1]南京大学医学院附属鼓楼医院肝胆与肝移植外科,南京210008 [2]安徽医科大学第一附属医院肝胆胰及移植外科,合肥230022

出　　处：《安徽医科大学学报》2024年第8期1361-1369,共9页Acta Universitatis Medicinalis Anhui

基　　金：国家自然科学基金面上项目(编号:82372834)。

摘　　要：目的探讨TBC1结构域家庭成员5(TBC1D5)在肝细胞癌(HCC)进展中的作用。方法利用蛋白质印迹实验(WB)、免疫组化(IHC)和实时荧光定量PCR(qPCR)分析TBC1D5在HCC肿瘤组织与癌旁组织间的表达量差异,构建相应的TBC1D5稳转肝癌细胞株;细胞计数试剂盒8、平板克隆实验和EdU实验检测细胞增殖能力变化;划痕实验和Transwell实验检测细胞迁移和侵袭能力,流式检测细胞周期变化和H2O2诱导的HCC细胞凋亡,最后通过WB检测敲低和过表达TBC1D5后对JAK/STAT通路的影响。结果WB、IHC和qPCR结果提示,HCC组织中TBC1D5在蛋白、mRNA水平表达量高于其对应癌旁组织(P<0.0001、P<0.01)。与对照组比较,敲低TBC1D5后HCC细胞增殖水平降低(P<0.05)、平板克隆集落数形成减少(P<0.001)、EdU阳性细胞比例下降(P<0.001)。划痕实验与Transwell实验结果显示,敲低TBC1D5后HCC细胞的迁移和侵袭能力相比于对照组降低(P<0.01)。敲低TBC1D5后HCC细胞相比于对照组,细胞周期减慢、抗凋亡能力降低(P<0.05、P<0.01)。与对照组相比,敲低TBC1D5使JAK与STAT蛋白磷酸化水平下降(P<0.01)并抑制JAK/STAT通路。结论TBC1D5在HCC中高表达,TBC1D5敲低后,HCC细胞的增殖、迁移和侵袭能力、细胞周期速率以及抗凋亡能力均降低,并且可能通过JAK/STAT通路影响HCC进展。Objective To investigate the role of Tre2-Bub2-Cdc161 domain family member5(TBC1D5)in the development of hepatocellular carcinoma(HCC).Methods Western blot(WB),Immunohistochemistry(IHC)and quantitative real-time PCR(qPCR)were used to verify the difference in TBC1D5 expression in clinical samples.The HCC cell lines MHCC97H and Hep3B were chosen to construct the knockdown model.The effects on cell proliferation were detected by cell proliferation assay,colony formation assay and EdU assay.Wound assay and Transwell assay were used to detect cell migration and invasion.Flow cytometry was used to detect the changes of cell cycle and H 2O 2-induced apoptosis of HCC cells.Finally,the effects of knockdown and overexpression of TBC1D5 on JAK/STAT pathway were detected by WB.Results The results of WB,IHC and qPCR showed that the expression of TBC1D5 in HCC tissues was higher at the protein level(P<0.0001)and mRNA level(P<0.01)than that in corresponding adjacent tissues.Compared with the control group,the proliferation level of HCC cells with TBC1D5 knockdown was decreased(P<0.05),the formation of plate colony number decreased(P<0.001),and the proportion of EdU-positive cells decreased(P<0.001).The results of scratch assay and Transwell assay showed that the migration(P<0.01)and invasion ability(P<0.01)of HCC cells after TBC1D5 knockdown were significantly lower than those in the control group.After TBC1D5 knockdown,the cell cycle of HCC cells was slowed down(P<0.05)and the ability to resist apoptosis was reduced(P<0.01)than those in the control group.Compare with the control group,knockdown of TBC1D5 decreased the phosphorylation level of JAK and STAT proteins and inhibit the JAK/STAT pathway.Conclusion TBC1D5 is highly expressed in HCC.After knocking down TBC1D5,the proliferation,migration and invasion ability,cell cycle rate and anti-apoptosis ability of HCC cells decreased and may affect HCC progression through the JAK/STAT pathway.

关 键 词：肝细胞癌 TBC1D5 增殖 侵袭 JAK/STAT通路 

分 类 号：R735.7[医药卫生—肿瘤]