Low-dose Nonylphenol Mediates Proliferation Of SW480 Cells Via GPR30 In Colon Cancer

Objective: To investigate the effect of low dose Nonylphenol(NP)on the proliferation of colon cancer SW480 cells and whether the effect was mediated by G protein-coupled estrogen receptor 30(GPR30).Methods: 1.CCK-8(Cell Counting kit-8)method was used to determine the effect of different concentrations of NP on the proliferation of human colon cancer cells SW480 in vitro and the optimal experimental concentration.2.The changes of cell cycle and apoptosis were detected by flow cytometry after the intervention of Estradiol E2,NP and GPR30 inhibitors G15+NP in SW480 cells,respectively.3.Immunofluorescence assay was used to detect the expression of GPR30 after E2 and NP acted on human colon cancer cells SW480.4.Western Blot was used to detect the expression changes of GPR30 after E2,NP and NP+G15 treated human colon cancer cells SW480,respectively.5.RT-PCR was used to detect GPR30 mRNA expression changes after E2,NP and NP+G15 acted on human colon cancer cells SW480,respectively.Results: The low dose NP promoted the proliferation of SW480 cells.Compared with the blank control group and the E2 control group,the cell viability of the low dose NP group increased,and the low dose promoted the proliferation of SW480 cells.Compared with the NP group,the cell viability of the NP+G15group decreased significantly;The proportion of cells entering the S phase in the low dose NP group increased compared with the blank control group and the E2 control group.Compared with the NP group,the proportion of cells entering the S phase in the NP+G15 group decreased;The apoptosis rate of cells in the low dose NP group decreased significantly compared with the blank control group and the E2 control group.meanwhile,compared with the NP group,the apoptosis rate of SW480 cells in the NP+G15 group increased;The immunofluorescence assay showed that the expression of GPR30 protein was higher after treatment with low dose NP compared with blank control group and E2 control group.Western-Blot experiment showed that the expression of GPR30 protein in low dose NP treatment group was higher than that in blank control group and E2 control group.Meanwhile,compared with the NP group,the expression of GPR30 protein decreased in the NP +G15 treatment group;RT-PCR experimental results showed that compared with blank control group and E2 control group,the expression of gpr30 mRNA in low dose NP treatment group was increased.The expression of GPR30 mRNA in the NP+ G15 treatment group decreased compared to the NP group.Conclusion: Low dose NP may significantly promote the proliferation of SW480 cells by upregulating the expression of GPR30.