Study On The Mechanism Of Long Non-coding RNA FAM230B Regulating The Proliferation,Migration,Invasion And Epithelial Interstitiallization Of Hepatocarcinoma Cells

Objective Primary liver cancer is the fourth most common malignant tumor and the second cause of cancer death in China,which seriously threatens the life and health of our people.Hepatocellular carcinoma（HCC）accounts for 70 90% of primary liver cancer.It has a high mortality rate,difficult diagnosis,and poor prognosis.Only 20% of newly diagnosed patients will be eligible for potentially curative therapies such as liver transplantation,surgical resection,and radiofrequency ablation.Despite liver resection is the most frequently applied treatment,about 70% of patients will relapse,and 30% will suffer tumor-related death within five years after surgeries.Revealing the underlying mechanism of HCC may be essential for higher therapy efficiency.Therefore,the study of critical factors that can inhibit HCC cell metastasis is helpful in improving the therapeutic effect and prognosis of HCC.However,HCC is a complex disease that many factors lead to the occurrence and development of malignant tumors.At present,its etiology and pathogenesis are not completely clear.In addition to the mutation or abnormal expression of protein-coding genes,the mutation and misregulation of noncoding RNA（nc RNA）,especially long-chain non-coding RNA（LncRNA）,seem to play an essential role in cancer.Long non-coding RNA FAM230 B was involved in tumor growth,cell differentiation,and apoptosis in some cancers.However,the association and influence between long non-coding RNA FAM230 B and HCC have not been reported.This study aims to clarify whether long non-coding RNA FAM230 B regulates the proliferation,migration,invasion,and Epithelial-to-mesenchymal transition（EMT）of hepatoma cells and its mechanism.Methods With normal para cancer tissues and liver cells L02 as controls,RT-qPCR was used to detect the expression of LncRNA FAM230 B in HCC tissues and cells Hep G2,Huh7,SMMC-7721,And analyze the relationship between the expression of LncRNA FAM230 B and the clinicopathological factors of 90 patients.The Star Base predicts that miR-1-3p is a potential downstream target of LncRNA FAM230 B.The dualluciferase report verifies the targeted regulation relationship of LncRNA FAM230 B to miR-1-3p.Divided HCC cells Hep G2 and Huh7 into control group,si-NC group,si-FAM230 B group,pc DNA group;pc DNA-FAM230 B group,FAM230B+miRNC group,and FAM230B+miR-1-3p group;Transfected the plasmids into each group of cells separately or jointly with Lipofectamine 2000;MTT method and the number of cell colonies were used to detect cell proliferation;the scratch test was used to detect cell migration;Transwell chamber test was used to detect cell invasion;western blot method was used to detect the expression of epithelial cadherin（E-cadherin）,neural cadherin（Ncadherin）,phosphatidylinositol 3-kinase（PI3K）,protein kinase B（AKT）;immunohistochemical staining was used to observe the expression of PI3 K and AKT in tumor tissues;the tumorigenesis experiment in nude mice was used to detect the development of transplanted tumors.Results Compared with adjacent tissues,the level of LncRNA FAM230 B in HCC tissues increased,the level of miR-1-3p decreased,and the levels of LncRNA FAM230 B and miR-1-3p showed a negative correlation;The level of LncRNA FAM230 B is positively correlated with tumor size,TNM stage,and microvascular infiltration（P<0.05）.Compared with L02,LncRNA FAM230 B was up-regulated in Hep G2,Huh7,SMMC-7721,and miR-1-3p were down-regulated（P<0.05）;LncRNA FAM230 B and miR-1-3p had binding sites,LncRNA FAM230 B directly targeted miR-1-3p,overexpression of LncRNA FAM230 B inhibited the expression of miR-1-3p（P<0.05）;Overexpression of LncRNA FAM230 B can up-regulate the levels of N-cadherin,PI3 K,and AKT,downregulate the levels of E-cadherin,promote cell proliferation,increase scratch healing and the number of invaded cells,and promote the growth of transplanted tumors（P<0.05）;And miR-1-3p can partially reverse the malignant phenotype of LncRNA FAM230 B on HCC cells.Conclusion LncRNA FAM230 B promotes the proliferation,migration,invasion,EMT,and the development of transplanted tumors in vivo via targeting miR-1-3p and activating the PI3K/AKT signaling pathway may play a role.