LINC02036在肝细胞癌中的表达及其参与肿瘤侵袭转移的分子机制  

Expression of LINC02036 in hepatocellular carcinoma and its molecular mechanism in tumor invasion and metastasis

作　　者：陈峰杰[1] 李仙仙 李素琴[1] CHEN Fengjie;LI Xianxian;LI Suqin(Department of Pathology,Henan Nursing Vocational College,Anyang 455000,China;Heji Hospital Affiliated to Changzhi Medical College,Changzhi 046000,China)

机构地区：[1]河南护理职业学院病理教研室,安阳455000 [2]长治医学院附属和济医院,长治046000

出　　处：《临床与实验病理学杂志》2024年第12期1293-1299,共7页Chinese Journal of Clinical and Experimental Pathology

基　　金：河南省卫健委科研项目(20202231)。

摘　　要：目的基于LINC02036探讨肝细胞癌(hepatocellular carcinoma,HCC)侵袭转移的机制。方法收集60对匹配的相邻非肿瘤组织和HCC组织,采用RNA测序确定差异表达的lncRNA。应用体外构建LINC02036敲低或过表达Huh7细胞,分别采用CCK-8、集落形成试验、Transwell法检测细胞的增殖和侵袭能力。通过Western blot检测上皮-间质转化(epithelial-mesenchymal transition,EMT)标记蛋白表达。此外,通过RNA下拉和质谱(MS)鉴定了LINC02036和TGF-β1之间的相互作用,并通过RNA免疫沉淀(RIP)进一步验证。结果通过RNA测序确定LINC02036在相邻正常组织和HCC组织中差异表达。LINC02036高表达与肿瘤最大径(P=0.025)、晚期TNM分期(P=0.028)以及微血管侵犯(P<0.001)显著相关。体外实验中,过表达LINC02036明显提高了细胞活力、侵袭和EMT(P<0.05),而敲低LINC02036基因则显示出相反的效果(P<0.05)。TGF-β1被鉴定为一种潜在的相互作用蛋白,并在HCC细胞裂解物中被LINC02036下拉。RIP分析验证了LINC02036和TGF-β1之间的相互作用。此外,放线菌素D分析显示TGF-β1过表达增强了LINC02036的RNA稳定性(P<0.05)。结论LINC02036在HCC中发挥致癌作用,促进肿瘤增殖和微血管侵犯,相关机制可能涉及TGF-β1增强其RNA稳定性。Purpose To study the mechanism of invasion and metastasis of hepatocellular carcinoma(HCC)based on LINC02036 expression.Methods A total of 60 pairs of the HCC tissues along matched adjacent non-tumor tissues were collected.The differential expression of lncRNA was confirmed by RNA sequencing.LINC02036 knockdown and overexpressing Huh7 cells were constructed in vitro,and the proliferation and invasion ability of the cells were detected by CCK-8,colony formation test,and Transwell method,respectively.The epithelial-mesenchymal transition(EMT)marker protein was detected by Western blot.In addition,the interaction between LINC02036 and TGF-β1 was identified by RNA pull down and mass spectrometry(MS),which was further validated by RNA immunoprecipitation(RIP).Results It was confirmed by RNA sequencing that LINC02036 was differentially expressed in adjacent normal tissues and HCC tissues.High expression of LINC02036 was significantly correlated with the largest tumor size(P=0.025),advanced TNM staging(P=0.028),and microvascular invasion(P<0.001).In vitro experiments,overexpression of LINC02036 obviously elevated cell viability,metastasis,invasion and EMT(P<0.05),while knockout of LINC02036 showed the opposite effects(P<0.05).TGF-β1 was identified as a potential interactive protein and pulled down by LINC02036 in HCC cell lysates.We verified the interaction between LINC02036 and TGF-β1 by RIP assay.Moreover,actinomycin D assay showed that overexpression of TGF-β1 enhanced the RNA stability of LINC02036(P<0.05).Conclusion LINC02036 plays a carcinogenic effect in HCC,promotes tumor proliferation and microvascular invasion,and the related mechanism may involve the enhancement of its RNA stability by TGF-β1.

关 键 词：肝细胞癌 TGF-Β1 LINC02036 侵袭 上皮-间质转化 

分 类 号：R735.7[医药卫生—肿瘤]