The Research Of The Function Of Runt Related Transcription Factor 3 On Lung Cancer Cell Line A549 In Vitro

Purpose: In this study,we used plasmid vector-mediated human RUNT-related transcription factor 3 to infect lung cancer cell line A549 through liposome in vitro and to detect the effect of gene on the proliferation,migration and apoptosis of lung cancer cells.To explore the effect of RUNX3 on the proliferation,migration and apoptosis of lung cancer,and to explore potential targets for targeted lung cancer.Method:(1)In this experiment,four groups of A549 cell lines were studied.Four groups were set up as experimental group,positive control group,negative control group and blank group.Runx3 plasmid and liposome were added into the experimental group Lipofectamine? 2000 was added to the positive control group and Lipofectamine?2000 was added to the negative control group.In the negative control group,lipofectamine?2000 was added to the control group and 1640 medium in the blank group.(2)The expression of RUNX3(EGFP-(EGFP-RUNX3)was transfected with Lipofectamine? 2000,and the plasmid was transfected with Lipofectamine? 2000(EGFP-NC).(3)The plasmid vector(eGFP-RUNX3)expressing the RUNX3 gene was transfected into lipofectamine? 2000(EGFP-NC)was set as positive controlgroup,plus Lipofectamine? 2000 plus negative control group,single serum-free medium 1640 was used as blank control group.(4)The expression of RUNX3 mRNA in different groups was detected by QPCR(6)Transwell chamber test was used to detect migration and invasion of cell lines in different groups;(7)Using flow cytometry(FCM);(3)Using CCK-8 kit to detect cell proliferation in different groups;Cytology was used to detect apoptosis in different groups.Result:(1)The Runx3 gene plasmid and the no-load plasmid were successfully extracted and transfected into the cell line.(2)The results of fluorescence quantitative PCR(QPCR)showed that the mRNA expression of Runx3 gene was the highest in the experimental group,and the expression of Runx3 gene was significantly higher than that of the Runx3 gene(P <0.01).(3)CCK-8 method was used to detect the proliferation of cells in each group(12h24h 48 h 72h 96h).The results showed that all the cells showed an increasing trend over time In the experimental group,the expression of RUNX3 OD(0.883± 0.15),the positive control group(1.638 ± 0.17),the negative control group(1.631 ± 0.12)and the blank group(1.788 ± 0.18)were significantly higher than those in the experimental group(P <0.01).(4)In the invasion experiment,the mean number of cells in the three visual field was randomly selected under the high magnification of 200 times.The experimental group,the positive control group,the negative control group and the blank group were(28 ± 3.33),(100 ±2.66),(110 ± 2.33),(180 ± 3.66),each experimental group were lower than the control group(P <0.01);(5)Annexin V / PI double staining cells in each group withered ratio of necrosis and apoptosis,the results shown in the experimental group,positive control group,negative control And the ratio of necrosis and apoptosis in the control group were respectively 16.25%,5.39%,5.152%,4.202%,the proportion of apoptosis and necrosis in the experimental group was significantly higher than the respective control group(P <0.01).Conclusion: In this study,it was proved that Runx3 gene could inhibit the proliferation of lung cancer cells in vitro and block the invasion and migration of lung cancer cells,leading to the apoptosis of lung cancer cells.This study provides a new idea and possible new molecular therapy target for molecular targeted therapy.