肝癌细胞中EP3受体亚型表达及其调控肝癌细胞生长的研究  

The EP3 receptor splice variants expression patterns in liver cancer cell and the relationship to the growth of liver cancer cell

作　　者：陈萌[1] 马娟[1] 夏树开[1] 汪亦品[1] 白小明[1] 张丽[1] 冷静[1] 

机构地区：[1]南京医科大学肿瘤中心生殖医学重点实验室病理学系,江苏南京210029

出　　处：《南京医科大学学报（自然科学版）》2013年第1期16-21,共6页Journal of Nanjing Medical University(Natural Sciences)

基　　金：国家自然科学基金(81172003);江苏高校优势学科建设工程资助项目(JX10131801021)

摘　　要：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：This experiment sought to investigate the EP3 receptor splice variants expression patterns in liver cancer cell lines and their functions in controlling hepatoma cell growth.Methods：WST assay was used to detect the growth of liver cancer cell induced by EP3 receptor agonist Sulprostone;Two-dimensional electrophoresis and mass spectrometry were used to detect the protein level in CCLP1 cells by EP3 receptor agonist sulprostone;Real-Time PCR was used to detect the expression of EP3 receptor splicing variants in liver cancer cell.Results：CCLP1 cells were treated with 10 μmol/L sulprostone for 24 h,the results of cell growth rate increased to 131.46%（P 0.01）detected by WST assay;CCLP1 cells were treated with 10 μmol/L Sulprostone for 24h,twodimensional electrophoresis and mass spectrometry were used to detect the protein level.The results showed that promoting cell proliferation,invasion and cell metabolism related proteins were increased,on the contrary that reducing the cell metabolism rate, slowing down the cell growth,invasion and inhibiting cell metabolism related proteins were reduced;EP3-4,EP3-5,EP3-6 and EP3-7 splice variants were observed in HUH-7,Hep3B,HepG2,HuCCT1 and CCLP1 cells.Conclusion：EP3 receptor promoted the proliferation of liver cancer cell by up-regulating proteins that promoting cell growth and metabolism at same time down-regulating proteins that inhibiting cell growth and metabolism.EP3-4,EP3-5,EP3-6 and EP3-7 splice variants were observed in HUH-7,Hep3B, HepG2,HuCCT1 and CCLP1 cells.

关 键 词：PGE2 EP3受体 剪接体亚型 肝癌 

分 类 号：R735.7[医药卫生—肿瘤]