前列腺素E2通过EP4受体调控甲状腺乳头状癌细胞增殖与侵袭的机制研究OA

Objective To investigate the role of prostaglandin E2(PGE2)in the pathogenesis of papillary thyroid carcinoma and its underlying mechanisms.Methods Human thyroid follicular cells(Nthy-ori 3-1)and papillary thyroid carcinoma cells(TPC-1)were cultured.The concentration of PGE2 in the cell culture supernatant was measured by ELISA.The expression of prostaglandin E2 receptor 4(EP4)in both cell lines was detected by Western Blotting and qRT-PCR.TPC-1 cells were treated with 0 μmol/L,1 μmol/L,2 μmol/L,or 5 μmol/L PGE2,cell viability was assessed using the CCK-8 assay,and EP4 protein expression was detected by Western Blotting.TPC-1 cells were treated with 0 μmol/L or 5 μmol/L PGE2,and cell proliferation and invasion capabilities were evaluated using the CCK-8 assay and Transwell assay,respectively.Furthermore,TPC-1 cells were treated with 0 μmol/L PGE2,5 μmol/L PGE2,5 μmol/L PGE2 combined with 5 μmol/L EP4 receptor agonist(receptor agonist 2),or 5 μmol/L PGE2 combined with 5 μmol/L EP4 receptor inhibitor(L-161982),and cell proliferation and invasion capabilities were subsequently measured using the CCK-8 assay and Transwell assay.Results The PGE2 content in the culture supernatant of TPC-1 cells was higher than that of Nthy-ori 3-1 cells(P<0.05).The mRNA expression level of the EP4 receptor in TPC-1 cells was higher than that in Nthy-ori 3-1 cells(P<0.05).The protein expression level of the EP4 receptor in TPC-1 cells was higher than that in Nthy-ori 3-1 cells(P<0.05).The cell viability in the 5 μmol/L PGE2 group was higher than that in the 0 μmol/L PGE2 group(P<0.05).A repeated-measures ANOVA was conducted to compare the relative proliferation rates of cells in the 0 μmol/L and 5 μmol/L PGE2 groups at 24,48,72,and 96 h,and the results showed that the differences in relative proliferation rates among different time points were statistically significant(P<0.05)and that the differences in relative proliferation rates between the two groups were statistically significant(P<0.05).The differences in the change trends of relative proliferation rates between the two groups were also statistically significant(P<0.05).The number of cells passing through the membrane in the 5 μmol/L PGE2 group at 24 h was higher than that in the 0 μmol/L PGE2 group(P<0.05).The relative expression levels of EP4 protein in the 2 μmol/L and 5 μmol/L PGE2 groups were higher than those in the 0 μmol/L PGE2 group(P<0.05).A repeated-measures ANOVA was performed to compare the relative proliferation rates of cells in the 0 μmol/L PGE2 group,5 μmol/L PGE2 group,5 μmol/L PGE2+5 μmol/L EP4 receptor agonist 2 group,and 5 μmol/L PGE2+5 μmol/L EP4 L-161982 group at 24,48,72,and 96 h,and the results showed that the differences in relative proliferation rates among different time points were statistically significant(P<0.05)and that the differences in relative proliferation rates among the groups were statistically significant(P<0.05).The differences in the change trends of relative proliferation rates among the groups were also statistically significant(P<0.05).The number of cells passing through the membrane in the 5 μmol/L PGE2+EP4 receptor agonist 2 group was higher than that in the 5 μmol/L PGE2 group(P<0.05),while the number of cells passing through the membrane in the 5 μmol/L PGE2+EP4 L-161982 group was lower than that in the 5 μmol/L PGE2 group(P<0.05).Conclusion PGE2 promotes proliferation and invasion of TPC-1 cells via the EP4 receptor.