肝细胞生长因子及其受体在常染色体显性多囊肾病囊肿衬里上皮细胞的表达研究

Expression of hepatocyte growth factor and its receptor in autosomal dominant polycystic kidney disease cyst-lining epithelial cells

目的 研究常染色体显性多囊肾病(ADPKD)囊肿衬里上皮细胞中肝细胞生长因子(HGF)及其受体c-MET的分布。方法 采用酶联免疫(ELISA)法测定ADPKD患者囊肿液、血清和原代培养ADPKD囊肿衬里上皮细胞上清液中HGF浓度。逆转录聚合酶链反应(RT-PCR)、原位杂交、Western印迹、免疫组织化学及计算机图像定量分析 HGF、c-MET mRNA和蛋白在囊肿衬里上皮细胞的表达。结果 ADPKD未透析患者囊液中HGF浓度为(8.61±0.07)ng/ml,显著高于血清HGF浓度(0.26±0.05)ng/ml(P<0.01)。原代培养的囊肿衬里上皮细胞上清液中也检测到HGF,浓度为(1.43±0.01)ng/ml。体外培养的ADPKD囊肿衬里上皮细胞中c-MET mRNA含量(75.69±12.36)与HGF mRNA含量(79.89±11.58)呈正相关(r=0.54,P<0.01)。ADPKD囊肿组织中囊肿衬里上皮细胞c-MET mRNA含量(114.75±36.24)与HGF mRNA含量(136.48±39.69)呈正相关(r=0.50,P<0.01)。ADPKD囊肿组织中囊肿衬里上皮细胞c-MET蛋白含量(101.68±21.06)与HGF蛋白含量(125.03±19.34)、培养的囊肿衬里上皮细胞c-MET蛋白含量(11.60±1.83)与HGF蛋白含量(18.46±1.71)均呈正相关(r=0.51,P<0.05;r=0.67,P<0.01)。结论 ADPKD囊肿衬里上皮细胞可合成和分泌HGF。HGF可调节囊肿衬里上皮细胞表达c-MET。

Objective To explore the autocrine mechanism of hepatocyte growth factor (HGF) and its receptor c-MET distribution in autosomal dominant polycystic kidney disease (ADPKD) cyst-lining epithelial cells. Methods The concentration of HGF was examined with ELISA in ADPKD patients' cystic fluid, serum and cultured media of cyst-lining epithelial cells. The expression of HGF and c-MET mRNA and protein in cyst-lining epithelial cells was detected by RT-PCR, in-situ hybridization, Western blotting, immunohistochemical analysis and computer image analysis. Results The concentration of HGF in ADPKD non-dialyzed patients'cystic fluid was much higher than that in ADPKD patients' serum [(8. 61±0. 07)ng/ml vs (0.26±0.05) ng/ml, P <0. 01]. The concentration of HGF in cultured media of ADPKD cyst-lining epithelial cells was (1. 43±0. 01)ng/ml. In ADPKD cyst-lining epithelial cells cultured in vitro, the expression level of c-MET mRNA (75. 69±12. 36) was positively correlated with HGF mRNA (79. 89±11. 58) (r = 0.54, P < 0. 01) . In cyst-lining epithelial cells in ADPKD cystic tissues, the expression level of c-MET mRNA (114. 75±36. 24) was positively correlated with that of HGF mRNA(136. 48±39. 69) (r = 0. 50, P< 0.01). In addition, the expression level of c-MET protein (101. 68±21. 06) of cyst-lining epithelial cells in ADPKD cystic tissues was positively correlated with that of HGF protein (125. 03 ±19. 34) ( r = 0. 51, P < 0.05) . The expression level of c-MET protein (11. 60±1.83) of ADPKD cyst-lining epithelial cells cultured in vitro was positively correlated with that of HGF protein (18. 46±1. 71) ( r = 0. 67, P < 0. 01). Conclusions ADPKD cyst-lining epithelial cells synthesize and secret HGF. HGF adjusts the expression level of c-MET in cyst-lining epithelial cells.