Experimental study on the effect of Notch signaling regulated by TMEM100 on proliferation,migration,invasion and apoptosis of gastric cancer cells
Objective To investigate the effects of TMEM100 on proliferation,migration,invasion and apoptosis of gastric cancer cells and its possible molecular mechanism.Methods The expression of TMEM100 in gastric cancer tissues was analyzed by GEPIA database,and the expression of TMEM100 in gastric cancer cell lines was detected by real-time fluorescence quantitative PCR(qPCR).Gastric cancer SGC-7901 cells were divided into control group(empty carrier group),TMEM100 overexpressed group(transfected with overexpressed vector pcDNA3.0-HA-TMEM100),TMEM100 overexpressed+Notch1 agonist group(VPA-d4).MTT,scratch assay,Transwell chamber and flow cytometry were used to detect cell proliferation,migration,invasion and apoptosis,respectively.Results The TCGA data showed that TMEM100 in gastric cancer tissues was significantly decreased compared with normal gastric tissues,the difference was statistically significant(P＜0.05).The expression of TMEM100 in GES-1 cells was 1.000±0.103.They were higher than AGS cells(0.693±0.091),MKN-45 cells(0.567±0.079),MGC80-3 cells(0.491±0.094)and SGC-7901 cells(0.415±0.0.84),and the difference was statistically significant(P＜0.05).Compared with empty carrier group(1.000±0.184),the expression of TMEM100 protein(2.648±0.249)and mRNA(3.462±0.328)were increased in TMEM100 overexpressed group(P＜0.05).The expression of Notch1 protein(0.417±0.103)and mRNA(0.424±0.091)were decreased in TMEM100 overexpressed group(P＜0.05).Compared with empty carrier group,the proliferation activity of TMEM100 overexpressed group was significantly decreased(P＜0.05).Compared with TMEM100 overexpressed group,the cell proliferation activity in TMEM100 overexpressed+VPA-d4 group was significantly increased(P＜0.05).Scratch test showed that the scratch healing rate of empty carrier group was(80.3±12.3)%,higher than that of TMEM100 overexpressed group(35.1±7.38)%.The scratch healing rate of TMEM100 overexpressed+VPA-d4 group was(79.2±12.6)%,which was lower than that of TMEM100 overexpressed group(P＜0.05).The results of Transwell assay showed that the number of transmembrane cells in empty carrier group was 233±30,which was higher than that in TMEM100 overexpressed group(72±19),and the difference was statistically significant(P＜0.05).Compared with TMEM100 overexpressed group,the number of transmembrane cells in TMEM100 overexpressed+VPA-d4 group increased to 257±35,and the difference was statistically significant(P＜0.05).The apoptosis rate was(3.6±0.3)%in the empty carrier group and(19.6±1.6)%in the TMEM100 overexpressed group,the difference was statistically significant(P＜0.05).Compared with TMEM100 overexpressed group,the apoptosis rate of TMEM100 overexpressed+VPA-d4 group decreased to(4.0±0.4)%,and the difference was statistically significant(P＜0.05).Conclusion The expression of TMEM100 is decreased in gastric cancer tissues and cells,and it may play a role in tumor inhibition in gastric cancer by inhibiting Notch1 expression.
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