Expression of LINC02036 in hepatocellular carcinoma and its molecular mechanism in tumor invasion and metastasis
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 col-lected.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 for-mation 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 spec-trometry(MS),which was further validated by RNA immuno-precipitation(RIP).Results It was confirmed by RNA se-quencing that LINC02036 was differentially expressed in adja-cent 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 micro vascular 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,acti-nomycin D assay showed that overexpression of TGF-β1 en-hanced the RNA stability of LINC02036(P＜0.05).Conclu-sion 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.
万方数据
维普
