角质细胞生长因子对囊肿衬里上皮细胞细胞周期及其调控蛋白表达的影响  

Effect of keratinocyte growth factor on cell cycle and regulatory protein of cyst-lining epithelia in autosomal dominant polycystic kidney disease

作　　者：孙田美[1] 戴兵[1] 梅长林[1] 刘沙勤[1] 沈学飞[1] 王文靖[1] 汤兵[1] 张树忠[1] 赵海丹[1] 宋吉[1] 

机构地区：[1]第二军医大学附属长征医院肾内科解放军肾病中心,上海200003

出　　处：《肾脏病与透析肾移植杂志》2003年第6期516-519,共4页Chinese Journal of Nephrology,Dialysis & Transplantation

基　　金：国家自然科学基金(30170901;30271523);上海市卫生系统百名跨世纪优秀学科带头人基金(97BR047); 上海市科委重大基础研究项目基金(02JC14029)

摘　　要：目的:研究角质细胞生长因子(KGF)对常染色体显性多囊肾病(ADPKD)囊肿衬里上皮细胞细胞周期及其调控蛋白的影响,以初步探讨ADPKD囊肿发生、发展的细胞学机制.方法:应用MTT法检测KGF对ADPKD囊肿衬里上皮细胞的促增殖作用;应用流式细胞仪观察KGF对ADPKD囊肿衬里上皮细胞细胞周期的影响;应用免疫组化结合病理图文检测KGF对囊肿衬里上皮细胞细胞周期调控蛋白Cyclin D1、P21wafl表达影响.结果:①KGF能明显促进ADPKD囊肿衬里上皮细胞增殖,影响细胞周期,使G0～G1期细胞减少,S期细胞增多;②体外培养的ADPKD囊肿衬里上皮细胞存在CyclinD1和P21wafl蛋白,经图文分析,50 ng/mlKGF刺激后,Cyclin D1蛋白表达(0.41±0.04)较对照组(0.30±0.01)明显增强,P21wafl的表达(0.32±0.02)较对照组(0.37±0.03)明显减弱(P＜.01).结论:CyclinD1和P21wafl是ADPKD囊肿衬里上皮细胞细胞周期G1期进展的调控蛋白;KGF可能通过刺激Cyclin D1蛋白的表达,抑制P21wafl蛋白的产生,从而进一步促进细胞通过G1～S调控点,致使ADPKD囊肿衬里上皮细胞明显增殖.Objective: To investigate the effect of keratinocyte growth factor (KGF)on the cell cycle and regulatory protein of cyst-lining epithelia in autosomal dominant polycystic kidney disease (ADPKD) and explore the role of KGF in the ADPKD pathogenesis and development. Methodology: The proliferative effect of KGF on cyst-lining epithelia was assessed by MTT assay. The effect of KGF on cyst-lining epithelia cell cycle were observed by flow cytometer. The effect on cell cycle regulatory protein cyclin D1 and P21waf1 expression were studied through immunohistochemistry and image-analysis system. Results: KGF could promote the proliferation, affect cell cycle and lead to G1~S phase arrest of cyst-lining epithelia. In vitro, experiment proved cyclin D1 and P21waf1 expression in cyst-lining epithelia. After stimulation by 50ng/ml KGF, the level of cyclin D1 expression (0.41±0.04) significantly increased compared with that in the control (0.30±0.01), however, the level of P21waf1 expression (0.32±0.02) obviously decreased compared with that in the control (0.37±0.03). Conclusion: CyclinD1 and P21waf1 were cell cycle regulatory protein in cyst-lining epithelia. KGF could stimulate cyclin D expression and inhibit P21waf1 expression, which facilitate passing through G1~S check point and lead to the proliferation of cyst-lining epithelia.

关 键 词：角质细胞生长因子 上皮细胞 细胞周期 调控蛋白 流式细胞仪 常染色体显性多囊肾病 

分 类 号：R692.1[医药卫生—泌尿科学]