LncRNA PCBP1-AS1 Aggravates The Progression Of Hepatocellular Carcinoma Via Regulating PCBP1/PRL-3/AKT Pathway

Hepatocellular carcinoma(HCC)is emerging as a high incidence common carcinoma and the third highest tumor-related mortality worldwide.The overall prognosis of patients with hepatocellular carcinoma is poor because of the reasons such as occult primary onset,rapid development,low differentiation,strong metastasis and invasion,and high postoperative recurrence rate.Although clinical therapeutic methods such as hepatectomy,liver transplantation,radiofrequency ablation,and microwave therapy for liver cancer have achieved good results,there is still a high recurrence rate and metastasis rate in patients with liver cancer after treatment.Therefore,it is urgent to find a better treatment method because of the high recurrence and metastasis of liver cancer after treatment.The pathogenesis of HCC is multifactorial due to the mutation of the tumor suppressor gene leading to the function weakening or loss,and activation of the oncogene.There are many comprehensive reasons for the formation of malignant tumors in the liver,such as the dysfunction of liver cell function,the dysregulation of the hepatic microenvironment,the massive destruction of the cellular matrix,and the imbalance between tumor angiogenesis and anti-angiogenesis.Therefore,it is crucial to find one or more key protein markers that are clearly and efficiently associated with the development or metastasis of liver cancer to predict the biological dynamics of the tumor.Long non-coding RNA(Lnc RNA)usually refers to RNA with more than 200 nucleotides that lack protein-coding capacity.A large number of studies have shown that lnc RNAs play multiple roles in the pathogenesis of hepatocellular carcinoma,which has certain potential for the exploration of markers for disease diagnosis.There is a potential cis regulatory relationship between the coding protein and Lnc RNA,that is,in the same chromosome locus,there are corresponding long-chain non-coding genes within 10 kb of the flanks of a certain protein coding gene,and the Lnc RNA has the potential to regulate the corresponding protein-coding genes.Poly(C)binding protein 1(PCBP1),also known as hn RNP E1,is located at the chromosomal locus 2p12-13,has three KH domains,and has a special molecular structure known as ???????,which provides a binding site for RNA polycytosine.A large number of studies have shown that lnc RNAs play multiple roles in the pathogenesis of hepatocellular carcinoma,which provides certain potential for early detection,diagnosis and treatment of hepatocellular carcinoma.Currently,there are no relevant studies on the specific mechanism of PCBP1 in HCC,and whether Poly(r C)-binding protein 1 antisense RNA 1(PCBP1-AS1)affects the protein PCBP1 still remains largely explored.In this study,we used chip technology to identify lnc RNA in the preoperative and postoperative plasma of patients with hepatocellular carcinoma,and found that PCBP1-AS1 showed an increasing trend in preoperative specimens and a decreasing trend in postoperative specimens.q RT-PCR was used to detect the expression level of RNA in HCC tissues and paracancer tissues in 109 cases.The results showed that PCBP1-AS1 was highly expressed in hepatocellular carcinoma tissues and relatively low in paracancer tissues.Furthermore,the clinicopathological data of 109 patients were analyzed,and the single-factor analysis showed that the relationship between the high and low expression of PCBP1-AS1 in vascular invasion,intrahepatic metastasis and tumor size was statistical significance.The abnormally high expression of PCBP1-AS1 promoted the rapid growth of the tumor,which was prone to intrahepatic and extrahepatic metastasis,and the survival time of the patients was significantly reduced.Furthermore,cell function experiments in vitro were carried out,and real-time fluorescence quantitative PCR was used to screen cell lines from HCC cell lines and normal L02 liver cell lines for the construction of PCBP1-AS1 overexpression and interference cell lines.It was found that PCBP1-AS1 could promote the proliferation of liver cancer cells by using the clone formation experiment,EDU and CCK8 experiments,and the cell scratch healing experiment,cell migration and invasion experiment showed that PCBP1-AS1 could promote the invasion and migration of liver cancer cells.In animal model experiments,we established subcutaneous xenotransplantation model and tail vein xenotransplantation model in nude mice,and the results showed that the overexpression of PCBP1-AS1 promoted the proliferation,migration and invasion of liver cancer cells,thus promoting the growth of HCC and lung metastasis in vivo.Similar experimental methods were used to detect the m RNA and protein expression levels of PCBP1.Both in vivo and in vitro experiments showed that PCBP1 inhibited the migration and invasion of hepatocellular carcinoma.Overexpression of PCBP1 reduced the migration,invasion and lung metastasis of tumor cells,which was completely contrary to the effect of PCBP1-AS1 on HCC.For the follow-up mechanism study,FISH experiment and ISH experiment were used to confirm that PCBP1-AS1 and PCBP1 were mostly located in the cytoplasm.Then,we found that PCBP1-AS1 had a strong binding with PCBP1 by RNA pull down,and RNA immunoprecipitation confirmed that PCBP1-AS1 and PCBP1 antibodies were enriched.We conducted cell sequencing on Hu H7-LV-NC and Hu H7-LV-PCBP1-AS1 cells,and the experimental results again confirmed that PCBP1-AS1 promotes the proliferation and metastasis of liver cancer cells,and the biological function of PCBP1-AS1 is closely related to the PI3K-Akt signaling pathway.However,western blotting was used to detect the positive correlation between PCBP1-AS1 and the expression levels of PRL-3 and p-AKT,which ultimately affected the AKT pathway.The over-expression of PCBP1-AS1 induced the activation of PRL-3-AKT signaling pathway.Thus,the high expression of PCBP1-AS1 could promote the proliferation and metastasis of HCC by regulating the PCBP1-PRL3-AKT axis.To sum up,this paper studied the important role of PCBP1-AS1 in the occurrence and development of hepatocellular carcinoma,and PCBP1-AS1 can inhibit the target protein PCBP1 to promote the proliferation and metastasis of HCC by regulating the PCBP1-PRL3-AKT axis.