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目的探讨内皮素受体(ETR)-B拮抗剂BQ788和磷脂酰肌醇-3激酶(PI3K)抑制剂LY294002对黑素瘤细胞株A375骨桥蛋白基因的表达。方法对照组在人A375细胞培养体系中加入100 nmol/L内皮素3,试验组除加入100 nmol/L内皮素3外,分别加入1μmol/L BQ788和20μmol/L、50μmol/L LY294002,采用RT-PCR和蛋白质印迹方法检测骨桥蛋白的mRNA和蛋白表达。结果ETR-B拮抗剂BQ788和PI3K抑制剂LY294002均能显著减少人恶性黑素瘤细胞株A375中骨桥蛋白mRNA和蛋白的表达,与对照组比较,差异均有统计学意义(P<0.05)。高浓度较低浓度LY294002对骨桥蛋白表达的抑制作用更明显(P<0.05)。结论内皮素3/ETR-B能通过激活PI3K信号通路上调骨桥蛋白的表达。
Objective To investigate the expression of Osteopontin gene of melanoma cell line A375 by endothelin receptor (ETR) -B antagonist BQ788 and phosphatidylinositol 3 - kinase (PI3K) inhibitor LY294002. Methods The control group was treated with 100 nmol / L endothelin 3 in the culture system of human A375 cells. In addition to 100 nmol / L endothelin 3, 1 μmol / L BQ788, 20 μmol / L and 50 μmol / L LY294002, respectively, The mRNA and protein expression of osteopontin were detected by PCR and Western blotting. Results Both EQ-8 antagonist BQ788 and PI3K inhibitor LY294002 could significantly decrease the expression of osteopontin mRNA and protein in human malignant melanoma cell line A375. Compared with the control group, the differences were statistically significant (P <0.05) . The inhibitory effect of LY294002 at higher concentration and lower concentration on osteopontin expression was more obvious (P <0.05). Conclusion Endothelin-3 / ETR-B up-regulates the expression of osteopontin by activating the PI3K signaling pathway.