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目的:研究肝癌细胞中EP3受体剪接体亚型的表达类型及其调控肝癌细胞生长的功能。方法:EP3受体激动剂Sulprostone处理肝癌细胞株观察其生长情况;二维电泳和质谱分析研究EP3受体激动与下游信号蛋白之间的关系;Real-TimePCR实验鉴定肝癌细胞株中EP3受体剪接体亚型的表达类型。结果:10μmol/L Sulprostone处理CCLP1细胞24 h后,细胞生长率上调了31.46%(P<0.01);给予CCLP1细胞10μmol/L Sulprostone处理24 h后,二维电泳和质谱实验的结果显示促进细胞增殖侵袭、与细胞代谢正相关的蛋白水平升高,降低细胞代谢速度、减慢细胞生长、抑制细胞侵袭转移的相关蛋白水平下降;Real-Time PCR实验检测肝癌细胞,结果表明肝细胞癌HUH-7、Hep3B、HepG2细胞及胆管细胞癌HuCCT1、CCLP1细胞表达EP3-4、EP3-5、EP3-6和EP3-7 4种剪接体亚型。结论:EP3受体通过上调促细胞生长代谢蛋白,下调抑制细胞生长代谢蛋白而发挥促进肝癌细胞增殖的作用。肝细胞癌HUH-7、Hep3B、HepG2细胞及胆管细胞癌HuCCT1、CCLP1细胞表达EP3-4、EP3-5、EP3-6和EP3-7 4种EP3受体剪接体亚型。
OBJECTIVE: To study the expression of EP3 receptor spliceosome subtypes in hepatocellular carcinoma cells and its function of regulating the growth of hepatocellular carcinoma cells. Methods: EP1 receptor agonist Sulprostone was used to treat hepatocellular carcinoma cell line to observe its growth. The relationship between EP3 receptor agonist and downstream signaling protein was studied by two-dimensional electrophoresis and mass spectrometry. Real-time PCR was used to identify the splicing of EP3 receptor in hepatocellular carcinoma cell line Body subtypes of expression types. Results: After treated with 10 μmol / L Sulprostone for 24 h, the cell growth rate was increased by 31.46% (P <0.01). After treated with 10 μmol / L Sulprostone for 24 h, the results of two-dimensional electrophoresis and mass spectrometry showed that the proliferation of CCLP1 cells was promoted Invasion, the positive correlation with cell metabolism increased protein levels, reduce cell metabolism, slow down cell growth, inhibition of cell invasion and metastasis related protein levels decreased; Real-Time PCR test of liver cancer cells, the results show that the hepatocellular carcinoma HUH-7 , Hep3B, HepG2 cells and cholangiocarcinoma HuCCT1, CCLP1 cells expressed four subtype of spliceosomes of EP3-4, EP3-5, EP3-6 and EP3-7. Conclusion: EP3 receptor can promote the proliferation of HCC cells by up-regulating the cell growth-promoting protein and down-regulating the cell growth and metabolism of protein. Hepatocellular carcinoma HUH-7, Hep3B, HepG2 cells and cholangiocarcinoma HuCCT1, CCLP1 cells expressed EP3-4, EP3-5, EP3-6 and EP3-7 EP3 receptor spliceosome subtypes.