The Effect Of Silencing ARID1A On The Pliferation Inhibition Of Gastric Cancer MGC-803 Cells

Objective This study used lentiviral infection technique to construct a human gastric cancer MGC-803 cell model with ARID1A gene silencing to investigate the effect of ARID1A gene silencing on the pliferation inhibition of human gastric cancer cells.Methods Experimental grouping:according to the ARID1A gene sequences provided by the Gen Bank database,three ARID1A target interference sequences were designed,named as KD1?KD2?KD3,negative control group(NC).Three ARID1A target interference sequence primers were annealed to form double-stranded DNA,and ligated and transformed into linearized GV493 vector.Virus packaging and determination T virus titer in 293 cells.Recombinant lentivirus was transfected into human gastric cancer MGC-803 cells to RT-PCR and Western blot detect cell ARID1A m RNA and protein expression levels.Cell proliferation,cell cycle,cell migration,cell invasion,apoptosis and other pliferation inhibition of ARID1A cells after gene silencing were studied.Results The results showed that the designed interference sequence was inserted into GV493vector correctly and sh RNA-ARID1A recombinant vector was constructed successfully.Lentivirus vectors were packaged in 293 T cells with a virus titer TU/m L8*10~8.RT-PCR and Westen blot results showed that lentiviral vector transfected ARID1A cells significantly reduced the m RNA and protein expression levels of the gene.After successful transfection,KD3 groups of cells were selected for cell proliferation,cell cycle,cell migration,cell invasion,apoptosis and other cell behavior experiments.Results After silencing the ARID1A,the cell proliferation experiment showed that the proliferation ability of KD3 group was enhanced compared with that of NC group,and its effect on the proliferation enhancement of MGC-803 cells was time-dependent,and the difference was statistically significant(P<0.05).The cell cycle experiment showed that KD3 group had more cells entering the S stage than NC group,23.05%±0.09%of the cells entering the S stage and 27.97%±0.38%of the cells entering the S stage in the KD group,indicating that the cell proliferation was active and the difference was statistically significant(P<0.05).The cell migration experiment showed that the migration ability of KD3 group was increased compared with NC group,the cell scratch was 24 hours,the average mobility of NC group was 29%±0.06%,and the average mobility of KD group was 47%±0.03%,the difference was statistically significant(P<0.05).The cell invasion experiment showed that KD3 group had 187±2.51 cells passing through,NC group had 110±1.52cells passing through,and KD3 group had more invasion ability.The apoptosis rate of KD3 group was 4.54%,that of NC control group was 25%,and that of KD3 group was significantly lower than that of NC group.Experiments show that ARID1A gene can inhibit cell proliferation,cell migration,cell invasion and promote apoptosis.Conclusions The proliferation,migration and invasion ability of gastric cancer MGC-803cells were enhanced after targeting silencing ARID1A genes in human gastric cancer cells;the apoptotic behavior was weakened,more cells entered the S stage,and the cell proliferation was active.