目的:探讨前列腺素E2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24h,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,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。HuccTl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PGE210μmol/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E2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 EPa-Ⅲ on cell growth in human cholangiocarcinoma cells. Methods:Human cholangiocarcinoma cells HuccT1 was used in this experiment. The cells were treated with various concentrations of selective EP3 agonist sulprostone. The cell viability was detected by WST-8. The transcription of EP3-Ⅲ in HuccT1 was detected by RT-PCR. HEK293 cells were transiently transfected with EP3-Ⅲ by lipofectamine 2000TM. The cell viability was detected in the transfected cells by WST-8. Results:Sulprostone induced a concentration-dependent increase in cell viability in HuccT1 cells. RT-PCR analysis showed EP3-Ⅲ mRNA expression in HuccT1 cells. When HEK293 cells were transiently transfected with EP3-Ⅲ, the cell viability was increased after exposured to PGE2 10μmol/L. Conclusion:These results demonstrate that EP3-Ⅲ is expressed in human cholangiocarcinoma cells ,which may play an important role in cell growth.