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.
出处
《肾脏病与透析肾移植杂志》
CAS
CSCD
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)
