Mechanism of nonylphenol promoting colorectal cancer cell proliferation by ac-tivating ERK signaling pathway via inhibiting ESR2 expression
Objective To investigate the effect of Xenoestrogen Nonylphenol(NP)on the proliferation of color-ectal cancer cells and its relationship with the activation of ERK pathway.Methods The expression of ERβ in colorectal cancer cells was analyzed by The Cancer Genome Atlas(TCGA)and Human Protein Atlas(HPA).The expression of ESR2 in COLO205 cells was detected by quantitative fluorescent polymerase chain reaction(qRT-PCR)after treatment with different concentrations of NP.Western blot was used to analyze the effect of NP on ERβ expression in COLO205 cells.Cells were divided into Control group,si-NC group,NP(10-6 mol/L)group,si-ESR2 group,and NP+si-ESR2 group.The expression of ESR2 was inhibited by using a small in-terfering RNA fragment(si-ESR2).After NP(10-6mol/L)was applied to COLO205 cells for 24 h,the prolif-eration and apoptosis of COLO205 cells were detected by CCK-8 and flow cytometry.The expression of apoptosis-related proteins(Bad,Bcl-2,and Cleaved caspase-3)and activation of ERK pathway were detected by Western blot.Results TCGA and HPA analysis showed that the expression of ERβ in CRC tissues was significantly lower than that in normal tissues(P＜0.001).The interference vector was constructed and transfected into the cells.The interference efficiency of si-ESR2-3# was more than 60％,and there was no significant difference in the si-NC control group.NP intervention significantly inhibited the expression of ESR2 and ERβ in COLO205 cells(F=27.791,＜0.001),and NP(10-6mol/L)significantly increased cell proliferation(F=107.3,P＜0.001)and inhibited cell apoptosis(F=51.1,P=0.006 8)compared with the Control group.The expression of Bad(F=46.56,P=0.003 2)and Cleaved caspase-3(F=18.04,P=0.003 5)were decreased,the expression of Bcl-2 was increased(F=32.58,P=0.005 4),and the expression of p-ERK was significantly increased(F=39.07,P=0.001 1).Compared with NP group,the viability and proliferation of cells in NP combined with si-ESR2 group were increased(F=107.3,P＜0.001),the apoptosis rate was significantly decreased(F=51.1,P=0.000 3),and the expression of Bad(F=46.56,P=0.000 9)and Cleaved caspase-3(F=18.04,P=0.001 9)were significantly decreased.The expression of Bcl-2(F=32.58,P=0.001 9)and p-ERK was sig-nificantly increased(F=39.07,P=0.008 1).Conclusion Nonylphenol can promote the proliferation of CRC cells by activating ERK pathway through ESR2.
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