The Expression And Function Of Long Non-coding RNA LINC01093 In Hepatocellular Carcinoma

ObjectiveTo investigate the disordered expression of long non-coding RNAs(lncRNAs)in hepatocellular carcinoma(HCC)tissues and corresponding adjacent non-tumor(ANT)liver tissues.And explore the correlation between the disordered expression of LINC01093 and clinical features as well as the function of LINC01093 in hepatocellular carcinoma,providing the theoretical basis for the hepatocellular carcinoma in early diagnosis,targeted therapy and prognosis analysis.MethodsHigh-throughput lncRNAs microarray was performed in 5 paired HCC tissues and ANT tissues.LINC01093,a novel lncRNA,was identified to be significantly down-regulated in HCC.Further,detecting the expression of LINC01093 in 104 pairs of HCC tissues and ANT tissues by quantitative real-time polymerase chain reactions(qRT-PCR),and elucidate the clinical significance of LINC01093.In vitro functional assays,conducting a stably transfected HepG2 cells lines withLINC01093 overexpression,and to explore the ability of proliferation,apoptosis,invasion,migration and the variety of cell cycle.The cell proliferation in Hep G2 cells after transfection with Linc01093 was explored by MTT.The cell cycle distribution and cell apoptosis rate in HepG2 cells after transfection with Linc01093 was explored by flow cytometry.The invasion and migration of Linc01093 in HepG2 cells after transfection with Linc01093 was detected by Transwell.Resulets(1)Microarray analysisA panel of long non-coding RNA transcripts were aberrantly expressed,1708 lncRNAs up-regulation and 2725 lncRNAs down-regulated(p<0.05).A total of 19 lncRNAs were p<0.01,of which12 lncRNAs were up-regulated and 7 lncRNAs were down-regulated.Bioinformatics analysis shown that LINC01093 was expressed in liver tissues specially and down-regulated significantly(p<0.01)in the microarray screening.(2)qRT-PCRUsing the qRT-PCR to further detect the expression of LINC01093 in 104 pair of HCC tissues and ANT tissues.LINC01093 was down-regulated in HCC tissues(p<0.001)as well as the results of microarray screening.(3)Clinical significanceThe HCC patients was divided into high-expression group and down-expression group according the median in the relative expression of LINC01093.In the basic demographic information of HCC patients,the cases proportion in the high-expression group and low-expression groupwas significantly different in HBV carriers(p=0.044)and HCC family history(p=0.006).In the clinic-pathological features of HCC patients,the cases proportion in the high-expression group and down-expression group was significantly different in tumor differentiation(p=0.001),whether the nodule(p=0.011)and BCLC staging(p=0.020).In routine detection of HBV test of HCC patients,the cases proportion in the high-expression group and down-expression group was significantly different in HBeAb(p=0.003).In the hematological index of HCC patients,the cases proportion in the high-expression group and down-expression group was significantly different in alpha fetoprotein(AFP)(p=0.030).In the immunohistochemical index of HCC patients,the cases proportion in the high-expression group and down-expression group was significantly different in CD34(p=0.005).(4)In vitro experiment,using the q RT-PCR to further detect the expression of LINC01093 in HCC cell of HepG2、Huh7、SMCC-7721 and normal liver cell of LO2,respectively.LINC01093 was significant down-regulated in HCC cell lines(p<0.001).(5)Studies on the function of cellThe ability of cell proliferation in HepG2 was decreased and rate of cell apoptosis was increased with overexpression of LINC01093,but not in invasion and migration(p>0.05).The variation of cell cycle had significant difference(p<0.05)with overexpression of Linc01093.The cells of G0/G1 phase was significantly increased,the cells of G2/M phase was increased but the cells of S phase was significantly decreased.ConclusionThe expression of LINC01093 is down-regulated in the tissues andcells of HCC.The ability of proliferation is declined and has an effect on the cell cycle with LINC01093 overexpression.It reveal that LINC01093 may play a role of tumor suppressor in the development of HCC,and it is expected to become a new molecular marker and therapeutic target for HCC.