Effects of interleukin-1 receptor Ⅱ on proliferation,migration and invasion of gastric cancer cells
Objective To investigate the effect of human interleukin-1 receptor Ⅱ(IL-1R2)on biological behavior of gastric cancer cells and its effect on blood vessels.Methods mRNA and protein levels of IL-1R2 in various gastric cancer cell lines(AGS,MGC-803,BGC-823,SGC-7901,HGC-27,HSC-39)and human gastric mucosa cells(GES-1)were detected by reverse transcriptase-polymerase chain reaction(RT-PCR)and Western blotting.MGC-803 and BGC-823 were selected to construct down-regula-ted and overexpressed IL-1R2 cell models.The effects of IL-1R2 on the proliferation,migration and inva-sion of gastric cancer cells were determined by cell counting kit-8(CCK-8)assay,scratch test and Tran-swell invasion test.The effects of IL-1R2 on cell cycle and apoptosis were investigated by cell cycle and apoptosis experiment.Vascular endothelial growth factor(VEGF)mRNA levels in MGC-803 and BGC-823 cells with down-regulated or overexpressed IL-1R2 were detected by RT-PCR.The cells in the down-regu-lated and overexpressed IL-1R2 groups and the control group were co-cultured with vascular endothelial cells(HMEC-1)to determine the effects of IL-1R2 on the proliferation and migration of HMEC-1 and the mRNA levels of VEGF and IL-6.The difference expression between the two groups was compared by t test.Analysis of variance(ANOVA)and Tukey trend test were used among the groups.Results RT-PCR and Western blotting showed that the mRNA and protein levels of IL-1R2 in gastric cancer cell lines(AGS,MGC-803,BGC-823,SGC-7901,HGC-27)were significantly higher than GES-1(mRNA:1.944± 0.310,4.777±0.320,2.214±0.100,3.235±0.121,4.809±0.190 vs.1.000±0.118,t=2.971,11.54,10.01,16.06,18.81,P＜0.05,P.＜0.01;Protein:2.010±0.151,4.445±0.362,1.988± 0.113,2.821±0.083,4.777±0.280 vs.1.000±0.087,t=5.973,11.54,10.01,16.06,18.81,P＜0.01,P＜0.01).CCK-8 assay,scratch test and Transwell invasion test suggested that IL-1R2 could promote the proliferation of gastric cancer cells(48,72,96 h in down-regulated group:0.298±0.028 vs.0.438±0.030,0.418±0.030 vs.0.592±0.026,0.599±0.025 vs.0.779±0.014,t=3.415,4.396,6.193,P＜0.05,P＜0.01;in the ovcrcxprcssion group at 72 and 96 h:0.745±0.051 vs.0.562±0.009,0.886±0.025 vs.0.790±0.015,t=3.577,3.306,P＜0.05,P＜0.01),migration(down regulation group:0.620±0.070 vs.1.000±0.080,t=3.587,P＜0.05;overexpression group:1.643±0.116 vs.1.000±0.067,t=4.810,P＜0.01)and invasion(down-regulation group:76.670± 4.631 vs.144.300±4.978,t=9.953,P＜0.01;in the overexpression group,195.700±4.807 vs.141.700±7.219,t=6.226,P＜0.01).The cell cycle experiment showed that the number of S phase cells increased in gastric cancer cells down-regulated by IL-1R2(32.700±0.879 vs.25.500±0.209,t=7.967,P＜0.01),but did not enter G2 and M phase.The results of RT-PCR showed that the content of VEGF mRNA in MGC-803 cells with down-regulated IL-1R2 was significantly lower than that in empty carrier control group(0.573±0.039 vs.1.000±0.011,t=3.721,P＜0.05).VEGF mRNA content in BGC-823 cells with overexpressed IL-1R2 was significantly higher than that in empty carrier control group(1.368±0.052 vs.1.000±0.084,t=3.715,P＜0.05).The co-culture experiment indicated that the invasion ability of HMEC-1 in MGC-803 cell group with down-regulated IL-1R2 was significantly decreased(55.330±3.756 vs.77.670±5.925,t=3.183,P＜0.05).The invasion ability of HMEC-1 in BGC-823 group overexpressing IL-1R2 was significantly enhanced(103.700±4.978 vs.80.330±3.383,t=3.877,P＜0.05).MGC-803 cells with down-regulated IL-1R2 were co-cultured with HMEC-1.VEGF mRNA content in HMEC-1 was lower than that in empty carrier control group(0.446±0.023 vs.1.000± 0.029,t=15.03,P＜0.01),and IL-6 mRNA level was higher than that in empty carrier control group.There was no significant difference(P＞0.05).When BGC-823 cells overexprcssing IL-1 R2 were co-cul-tured with HMEC-1,VEGF mRNA in HMEC-1 was higher than that in empty carrier control group(1.896±0.236 vs.1.000±0.048,t=3.717,P＜0.05),and IL-6 mRNA level was higher than that in empty carrier control group.There was no significant difference(P＞0.05).Conclusion IL-1R2 can promote the proliferation,migration and invasion of gastric cancer cells,and promote the formation of tumor blood vessels.
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