前列腺素E_2受体EP3-Ⅲ对胆管上皮癌细胞生长的影响

The effect of prostaglandin E_2 receptor EP3-Ⅲ on cell growth in human cholangiocarcinoma cells

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 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.