The Effect Of Knockdown OLFML2A Gene On The Biological Behavior Of Hepatocellular Carcinoma Cells

Objective:1.Detection of OLFML2 A gene expression in liver cancer cell lines;2.Construction of OLFML2 A RNAi Lentivirus;3.Detection of OLFML2 A RNAi Lentivirus infection BEL-7404 cells,cell proliferation,cell cycle and apoptosis,initially identified OLFML2 A gene on the biological behavior of liver cancer cells.Method:1.Construction of OLFML2 A RNAi Lentivirus;2.GFP green fluorescence was observed by fluorescence microscope to determine the infection efficiency.The changes of OLFML2 A gene expression after infection with lentivirus were analyzed by RT-qPCR and Western-bolt.3.The BEL-7404 cell line in the logarithmic growth phase was divided into two groups: Lentiviral vector and lentivirus group,which were infected with lentivirus empty vector and lentivirus respectively.The growth rate of liver cancer BEL-7404 cells was detected by Celigo method;MTT The cell doubling rate was detected by the method;the change of cell proliferation cycle and apoptosis was detected by FACS.Result:1.Successfully constructed RNAi lentivirus of OLFML2 A gene.2.Human hepatocellular carcinoma cell line BEL-7404 was successfully infected with OLFML2 A RNAi lentivirus.Microscopically,green fluorescence was observed with an infection rate of 80%;OLFML2A mRNA levels were significantly decreased after infection(p=0.006,p<0.05).The expression of OLFML2 A gene was significantly reduced(p<0.01),and its knockdown efficiency was 68.6%.3.BEL-7404 cells infected with lentivirus: Compared with the lentivirus empty vector group,the growth rate of BEL-7404 cells in the lentivirus group was continuously inhibited(p=0.001,p<0.05);on the 5th day after cell infection,L1 decreased cell number in G1 phase(p<0.05),no significant change in the number of cells in S phase,significantly increased in G2/M phase cells(p<0.05);3 days aftercell infection,Lentivirus The number of apoptotic cells was significantly increased in the group(p<0.05).Cell proliferation was detected by MTT assay: The cell proliferation rate in the lentivirus group was significantly decreased(p<0.05),and the difference was statistically significant.Conclusion:The OLFML2 A gene RNAi lentivirus significantly inhibits the expression of OLFML2 A gene,inhibits the proliferation of human hepatoma BEL-7404 cells,promotes their apoptosis,and affects their cell cycle.