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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、P21 waf1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Cyclin D1和 P21waf1蛋白,经图文分析, 50 ng/ml KGF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,P21waf1的表达(0.32±0.02)较对照组(0.37±0.03)明显减弱(P<0.01)。 结论:Cyclin D1和 P21waf1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P21waf1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 the regulatory proteins of cyst-lining epithelial cells in autosomal dominant polycystic kidney disease (ADPKD), so as to investigate the cytological mechanism of ADPKD cyst. Methods: The effect of KGF on proliferation of ADPKD cyst-lining epithelial cells was detected by MTT assay. The effect of KGF on the cell cycle of ADPKD cyst-lining epithelial cells was observed by flow cytometry. The effect of KGF on cyst-lining epithelium Cell cycle regulatory proteins Cyclin D1, P21 waf1 expression. Results: ① KGF significantly promoted the proliferation of epithelial cells lining the lining of ADPKD cysts, affecting the cell cycle, decreasing the number of G0 ~ G1 phase cells and increasing the number of S phase cells. ② The Cyclin D1 and P21waf1 proteins were found in the lining epithelial cells of ADPKD in vitro. (0.41 ± 0.04) was significantly higher than that of the control group (0.30 ± 0.01), P21waf1 expression (0.32 ± 0.02) was significantly lower than that of the control group (0.37 ± 0.03) (P < 0.01). CONCLUSIONS: Cyclin D1 and P21waf1 are regulators of cell cycle G1 phase progression in ADPKD cyst-lining epithelial cells. KGF may inhibit the production of P21waf1 protein by stimulating the expression of Cyclin D1 and further promote the cells to pass G1-S regulatory sites, resulting in ADPKD Cyst lining epithelial cells significantly proliferated.