The Effect Of Silencing TGM2 Expression On The Growth And Proliferation In MGC803 Cells

Objectives: Gastric cancer is a common malignant tumor which is a serious threat to people’s health and life.In order to understand its pathogenesis,the group used quantitative proteomics technology in the early stages of the study to screen for Tissue Transglutaminase(TGM2)proteins from different stages of gastric mucosal cancer to further understand the protein’s role in the development of gastric cancer.Mechanism,It can provide experimental data for elucidating the molecular mechanism of gastric cancer.Methods: 1.The TGM2 interference vector was constructed to transfect into gastric cancer MGC803 cells.After screening,the MGC803 cell line of TGM2 low expression was established.2.The expression of TGM2 protein was detected by Western-blot and cell immunofluorescence in MGC803 cells.3.The cell growth curve,plate cloning and soft agar colony forming experiment and flow cytometry were used to observe the effect of TGM2 low expression on growth and proliferation,cycle and apoptosis in MGC803 cells.Results:1.The pcDNA6.2-GW/Em GFP-Sin-TGM2 interference plasmid was successfully constructed.2.The results of cell growth curve showed that the growth rate of MGC803-Sin-TGM2 cells was significantly lower than that of the control group MGC803 and MGC803-pcDNA6.2 cells,and the low expression of TGM2 in MGC803 cells inhibited the growth and proliferation of cells(P<0.01).3.The results of plate clone formation experiment showed that the clones number of MGC803-Sin-TGM2 cells and the control group of MGC803 and MGC803-pcDNA6.2 cells were96±12,282±25 and 294±38.The clones number of MGC803-Sin-TGM2 cell significantly decreased compared with the control group and the clone size was small(P<0.01).The results showed that the colony formation ability of MGC803 cells was decreased significantly after low expression of TGM2 protein.4.The soft agar colony formation assay showed that the colonies number of MGC803-Sin-TGM2 cells and the control group of MGC803 and MGC803-pcDNA6.2 cell were 124±35,542±54 and 568±66.The colony formation of MGC803-Sin-TGM2 cell were significantly reduced,and the clone size was small(P<0.01).The colony forming ability of MGC803 cells decreased significantly after low expression of TGM2 protein.5.The flow cytometry showed that the percentage of S phase cells was 15.77±2.14,the percentage of G1 phase cells was 82.96±6.46,and the cell proliferation index was17.05 in MGC803-Sin-TGM2 cells.The percentage of S stage cells was 42.15±3.24,the percentage of G1 phase cells was 45.41±2.42,and the cell proliferation index was 54.60 in MGC803 cells.The percentage of S stage cells was 38.95±3.25,the percentage of G1 phase cells was 46.58±2.56,and the cell proliferation index was 53.42 in MGC803-pcDNA6.2 cells.The results showed that the proportion of S phase cells decreased significantly,the proportion of G1 phase cells increased and the cell proliferation index decreased after the low expression of TGM2 in MGC803 cells(P<0.01).Meanwhile,the apoptotic rate of MGC803-Sin-TGM2 cells,MGC803 and MGC803-pcDNA6.2 cells were respectively 35.94±3.45,18.28±2.56 and 12.84±2.1351.The results showed that the apoptosis rate of the cells increased after the low expression of TGM2 in MGC803 cells(P<0.01).Conclusions: The low expression of TGM2 reduced the proliferation index,increased cell apoptosis and inhibited the growth and proliferation of MGC803 cells.