| Hepatocellular carcinoma(HCC)is one of the most malignant tumors in the world.The obvious characteristics of HCC include hard to diagnose in the early stage,extensive invasion of hepatic surgery,adverse effects of chemotherapies,easy to be resistant to immunotherapies,and worse prognosis for OS(overall survival)or DFS(disease-free survival).Currently,the mechanism of HCC has not been revealed in detail yet.With the development of NGS(next-generation genome sequencing)and gene chip,it has been accessible for researchers to discern some new non-coding gene transcripts and study their functions in detail.Long non-coding RNA is usually defined as the kind of transcripts longer than 200 nucleotides without the ability of coding proteins.Recently,lnc RNAs are found to play an important role in the functions of tumor genesis,proliferation,invasion and migration,etc.Firstly,we have found a new lnc RNA FBXL19-AS1 which has not been reported in HCC,by gene chip sequencing of HCC tissues.We also find that lnc RNA FBXL19-AS1 is highly expressed in HCC,so we have designed this study to research the functions and mechanism of lnc RNA FBXL19-AS1.AimsTo elaborate the relationship between expression of lnc RNA FBXL19-AS1 and prognosis of HCC patients.MethodsCompare the expression of lnc RNA FBXL19-AS1 between cancer foci and para-cancer tissues or normal liver tissues,based on the public data of TCGA and GTEx.Make Kaplan-Mieir survival analyses by classifying the patients into lnc RNA FBXL19-AS1 high expression group and low expression group.Utilize RT-q PCR to analyze the expression of lnc RNA FBXL19-AS1 in different HCC cell-lines.ResultsIt is found that lnc RNA FBXL19-AS1 is more highly expressed in HCC cancer than para-cancer tissues or normal liver tissues,and lnc RNA FBXL19-AS1 high expression group has worse survival than low expression group.There are different expressions of lnc RNA FBXL19-AS1 in different HCC cell-lines.Conclusionlnc RNA FBXL19-AS1 is highly expressed in HCC tissues and is connected with worse survival of HCC patients.lnc RNA FBXL19-AS1 may develop as a kind of early diagnosis marker for HCC or the potential target site of immunotherapies.AimsTo interrogate the influence of the knocked-down expression of lnc RNA FBXL19-AS1 on the proliferation,apoptosis,invasion and migration of HCC.MethodsTest the changes of CCK8,EDU,Flow Cytometry(apoptosis,cell cycle),colony formation,wound healing experiment,Transwell(invasion,migration)and etc,after knocking-down the expression of lnc RNA FBXL19-AS1.Test the Subcutaneous tumors of HCC in nude mice(BCLB/C)after the HCC cells transducted by lenivirus with sh RNA.Utilize the FISH and PARIS? Seperation kit to analyze the sub-cellular location expression of lnc RNA FBXL19-AS1.ResultsIt is found that knockdown of lnc RNA FBXL19-AS1 can greatly deter the proliferation,cell-cycle,invasion and migration of HCC cell-lines.Meanwhile,it can also enhance the apoptosis of HCC.The proliferation of HCC in nude mice is also suppressed in vivo.Besides,the lnc RNA FBXL19-AS1 is high expressed in nucleus.Conclusionlnc RNA FBXL19-AS1 is vital important in the proliferation,apoptosis,invasion and migration of HCC both in vitro and in vivo.AimsTo annotate the regulatory mechanisms of lnc RNA FBXL19-AS1 in the proliferation,apoptosis,invasion and migration of HCC.MethodsSearch the specific proteins which can be bound with lnc RNA FBXL19-AS1 by CHIRP-MS test in Hep3 B.Screen the meaningful genes changing with the knockdown of lnc RNA FBXL19-AS1 in Hep-LM3 by RNA-Sequencing.Test the changing expressions of proteins connected with apoptosis by Western Blot.Figure out the relationship between lnc RNA FBXL19-AS1 and target genes or proteins via RT-q PCR,Western Blot,and RIP.ResultsIt is found that lnc RNA FBXL19-AS1 can be bound with protein Hn RNPA2B1 exclusively.There are in all 92 genes with changing expressions after knockdown of lnc RNA FBXL19-AS1,including 68 genes up-regulated and 24 genes down-regulated.We found a very meaningful gene(Fold changes 2.58,p<0.05),namely,dusp1,which is dual specificity phosphatase 1 and plays a very important role in MAPK pathway.To test the relationship of lnc RNA FBXL19-AS1,Hn RNPA2B1 and DUSP1,RT-q PCR proved that lnc RNA FBXL19-AS1 competes with DUSP1 in expression;DUSP1 is regulated by Hn RNPA2B1;There is no co-regulation between expressions of Hn RNPA2B1 and lnc RNA FBXL19-AS1.RIP displayed that Hn RNPA2B1 can cooperate with lnc RNA FBXL19-AS1 and m RNA(dusp1),respectively.Besides,proteins p53 、 Caspase3 、 Caspase8 、 Caspase9 can also be up-regulated by knocking-down of lnc RNA FBXL19-AS1.ConclusionLnc RNA FBXL19-AS1 can regulate the proliferation of HCC via Hn RNPA2B1/DUSP1/MAPK axis.It would be a great potential target site of diagnosis and immunotherapies of HCC in the future. |
PDF Full Text Request