Heatr-1 Gene Correlation Studies In Proliferation Of Gastric Cancer

Objective: A gastric cancer cell line AGS was divided into two groups: shCtrl(control group)and shHEATR1(target group).HEK293 cells were transfected with HEATR repeat containing-1 gene After lentivirus infection and knocking out the gene of interest,we observed whether the cell proliferation between normal two groups was inhibited.Method: Based on the nucleotide sequence of the mRNA of HEATR-1 searched in the gene bank,the lentiviral vector of target gene was designed and constructed.The target cells in good growth condition were cultured,and the experimental conditions were designed according to the pre-lentivirus infection experimental results,and the infection was performed formally.In the case of fluorescently labeled lentivirus infection,the expression of GFP was observed under a fluorescence microscope with reference to the pre-experiment time point of infection.The fluorescence reached about 70-80%,and the cell confluence reached 80%.The cells were collected for further experiments.The expression of AGS gene in gastric cancer cell line was detected by PCR and confirmed to be highly expressed in gastric cancer cells.The expression of HEATR-1 mRNA was detected by Q-PCR after the lentivirus infected target group.The effects of HEATR-1 knockdown on cell proliferation were evaluated by MTT assay in shCtrl(control group)and shHEATR1(target group),and proliferation of gastric cancer tissue was explored.Results:Control and purpose 72 hours after infection with lentivirus target cells fluorescence observation under a microscope,the results showed that the cell infection efficiency of more than 80% of the cells were normal.Fluorescent quantitative PCR was used to detect the gene knockdown of HEATR1 gene shCtrl and shHEATR1(shCtrl?Ct mean = 6.48,shHEATR1?Ct mean = 9.73,the expression abundance was 1.000 ± 0.014,0.105 ± 0.005,See Table 1).During the proliferative phase,exogenous MTT was removed and reduced to water-soluble blue-violet needle Formazan crystals and precipitated in the cells.The results of MTT assay showed that shHEATR1 cells had a more slow day5 proliferation compared with shCtrl group(P <0.05)).Based on the above experimental results,it was confirmed that interfering with the target gene HEATR1 can inhibit the proliferation of gastric cancer cell AGS Conclusion:Interfere with gene HEATR1 on gastric cancer cell AGS inhibit its proliferation.