Effect And Mechanism Of LncRNA HAND2-AS1 On Invasion Metastasis And Proliferation Of Hepatocellular Carcinoma

Objective:This project intends to explore the expression of lncRNA HAND2-AS1 in Hepatocellular carcinoma(HCC)cell lines.Reveal the effect of lncRNA HAND2-AS1 on the invasion,metastasis and proliferation of HCC cell lines.Uncover its mechanism in the initiation and development of HCC,so as to bring a new target for the clinical treatment of HCC.Methods:1 Under the premise that the normal liver cell line L02 was used as a control,RT-PCR was used to detect the expression level of lncRNA HAND2-AS1 in a variety of HCC cell lines.2 Two HCC cell lines with the lowest expression of lncRNA HAND2-AS1 in the above test results were selected for cell transfection.Then three groups were named:(1)the blank group without any intervention(blank group);(2)the control group(CON group)transfected with empty plasmid;(3)an experimental group with an overexpression plasmid(HAND2-AS1 group).The transfection efficiency was tested by RT-PCR.The wound healing assay,transwell chamber assay and MTT assay were used to detect the migration,invasion and proliferation capabilities of the above three groups of cell lines.3 Western blot assay was used to detect the contents of PI3 K and AKT in three distinct groups of the above two cell lines.4 Cell lines overexpressing HAND2-AS1 were treated with AKT activator and classified as AKT group.PI3K-AKT inhibitor LY294002 was added to the AKT group to classify it as the LY294002 group.The migration,invasion and proliferation abilities of these two groups of cell lines were tested respectively.Results:1 Compared with the normal liver cell line L02,the expression of lncRNA HAND2-AS1 in HCC cell lines Hep G2,SMMC-7721,Hep3 b,PLC,Huh-7,MHCC-97 H,MHCC-97 L was significantly decreased.Compared with other HCC cell lines,lncRNA HAND2-AS1 has lower expression in SMMC-7721 and MHCC-97 H cell lines.2 There was no difference in migration,invasion and proliferation of HCC cell lines between blank group and CON group.Compared with the blank group and the CON group,the migration,invasion and proliferation of HCC cell lines in the HAND2-AS1 group were significantly reduced.3 There was no difference in PI3 K and AKT in HCC cell lines between blank group and CON group.Compared with the blank group and the CON group,the expression of PI3 K and AKT protein in the two HCC cell lines in the HAND2-AS1 group was significantly decreased.4 Compared with the HAND2-AS1 group,the cell line migration,invasion and proliferation ability of the AKT group was significantly enhanced,and when PI3 K inhibitors were added,this enhancement effect was weakened.Conclusion:1 The expression of lncRNA HAND2-AS1 in HCC cell lines was significantly reduced,and it played an important tumor suppressor effect by regulating the PI3K-AKT signaling pathway.2 Overexpression of lncRNA HAND2-AS1 can significantly reduce the migration,invasion and proliferation of HCC cell lines.