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目的:探讨前列腺素E_2受体EP3-Ⅲ对胆管上皮癌细胞生长的影响。方法:采用常规培养的人胆管上皮癌细胞株HuccT1,分别给予不同浓度的EP3受体激动剂sulprostone(0、1、5、10μmol/L)作用24 h,用WST-8法检测细胞的活性。用RT-PCR方法检测HuccT1细胞表面EP3-ⅢmRNA水平的表达。用细胞瞬时转染方法使HEK293细胞表达EP3-Ⅲ蛋白,并用WST-8法检测PGE_2对转染HEK293细胞生长的影响。结果:WST-8检测发现,随着sulprostone作用浓度的升高,HuccT1的细胞活性呈浓度梯度依赖性增加。RT-PCR检测发现,HuccT1细胞有EP3-ⅢmRNA表达。HEK293细胞瞬时转染EP3-Ⅲ后,WST-8检测发现转染细胞在PGE_2 10μmol/L作用下细胞活性明显增加。结论:人胆管上皮癌细胞株HuccT1表面有前列腺素E_2受体EP3-Ⅲ表达,EP3-Ⅲ对细胞生长有显著的促进作用。
Objective: To investigate the effect of prostaglandin E2 receptor EP3-III on the growth of bile duct epithelial carcinoma cells. METHODS: Conventional cultured human bile duct epithelial carcinoma cell line HuccT1 was used to treat different concentrations of EP3 receptor agonist sulprostone (0, 1, 5, and 10 μmol/L) for 24 h. The activity of the cells was measured by WST-8 assay. The expression of EP3-III mRNA on the surface of HuccT1 cells was detected by RT-PCR. EP3-III protein was expressed in HEK293 cells by transient transfection method, and the effect of PGE2 on the growth of transfected HEK293 cells was detected by WST-8 assay. RESULTS: WST-8 detection revealed that the concentration of sulprostone increased the concentration of HuccT1 in a concentration-dependent manner. RT-PCR detection revealed that the HuccT1 cells had EP3-III mRNA expression. After transient transfection of EP3-III in HEK293 cells, WST-8 assay showed that the activity of cells transfected with PGE 2 at 10 μmol/L was significantly increased. Conclusion: The expression of prostaglandin E2 receptor EP3-III is expressed on the surface of human bile duct epithelial cancer cell line HuccT1. EP3-III has a significant effect on cell growth.