羧酸类化合物DJ5对人肾囊肿衬里上皮细胞的增殖抑制作用

目的：观察羧酸类化合物DJ5对人肾囊肿衬里上皮细胞WT9—12的增殖抑制作用并探讨其作用机制。方法：Mvr法检测细胞增殖，LDH释放实验检测细胞毒性，流式细胞术检测细胞周期和细胞凋亡，Westernblot检测周期相关蛋白P2l、cyclinA，凋亡蛋白队RP，通路蛋白Akt、Fox03a蛋白表达。结果：实验表明100p,mol／L的DJ5作用WT9—12细胞72h，细胞增殖抑制率为59．2％，DJ5使细胞周期阻滞在S期并促进细胞早期凋亡。DJ5可上调WT9—12细胞周期抑制蛋白P2l表达、下调cyclinA表达；使PARP活性片段89kD表达明显增加。DJ5可下调磷酸化Akt、磷酸化Fox03a表达、上调Fox03a表达。结论：DJ5通过阻滞细胞周期、促进细胞凋亡及抑制Akt—Fox03a通路蛋白发挥抑制WT9—12细胞增殖作用。

Anti-proliferative Effect of Carboxylic Acid Compounds DJ5 on Autosomal Dominant Polycystic Kidney Disease Cystic Lining Epithelial Cells

Objective：To observe the anti - proliferative effect of Carboxylic Acid Compounds DJ5 on autosomal dominant polycystic kidney disease cystic lining epithelial cells and to investigate the underlying molecular mechanism. Methods： ADPKD cystic lining epithelial （WT9 - 12） cells were stimulated by DJ5 with different concentrations. After treatment, MTY method was porformed to detect the level of proliferation, LDH release method was performed to detect the cytotoxicity ;flow cytometry was used to determine the cell cycle distribution and the apoptosis rate, western blotting was used to detect the protein expressions of P21, cyclin A, PARP, Akt, phosphorylated Akt, FoxO3a, phosphorylated FoxO3a. Results：Treatment of WT9 -12 cells with DJ5 resulted in a time- de pendent and dose - dependent inhibition of cell proliferation. Treatment of WT9 - 12 cells with DJ5 100 μmol/L for 72 h, the inhibi ton rate was 59.2%. DJ5 can make WT9 - 12 cells accumulated in S phase , up - regulate the expression of P21 CIP/WAF1 and down regulate cyclin A. DJ5 100 trmol/L can make WT9 - 12 apoptosis, and the expression of PARP with 89kd is enhanced. Phos phorylated Akt, phosphorylated FoxO3a is down -regulation and FoxO3a is up - regulation in DJ5 groups. Condusion：DJ5 inhibits the proliferation of WT9 -12 cells through accumulating WT9 -12 cells in S phase, promoting apoptosis and regulat!ng Akt -FoxO3a signal pathway.