刺参黏多糖抑制胰腺癌SW1990细胞增殖的机制研究

The mechanism of inhibitory effect of SJAMP on the proliferation in human pancreatic cancer SW1990＇s cells

目的 探讨刺参黏多糖抑制胰腺癌SW1990细胞增殖的可能机制.方法 应用8 mg/ml刺参黏多糖干预胰腺癌SW1990细胞24、48、72 h,采用RT-PCR法检测细胞Hippo信号通路成员YAP、MST1及相关基因Survivin、Caspase-9、TEAD1 mRNA的表达；蛋白质印迹法检测细胞YAP、磷酸化YAP（pYAP）蛋白的表达.结果 以对照组细胞的表达量为1,刺参黏多糖干预SW1990细胞48 h后细胞的YAP 、TEAD1、Survivin、Caspase-9、MST1 mRNA表达量分别为0.27、0.31、0.26、4.06、7.06,其中YAP、TEAD1、Survivin mRNA表达下调,Caspase-9、MST1 mRNA表达上调,且呈时间依赖性的变化趋势,与对照组的表达量差异均具有统计学意义（P ＜0.05或＜0.01）.对照组细胞YAP、pYAP蛋白的表达量分别为0.705±0.021、0.430±0.043,刺参黏多糖干预SW1990细胞48 h后YAP、pYAP的表达量分别为0.545±0.071、0.652±0.025,其中YAP表达下调,pYAP表达上调,且呈时间依赖性的变化,与对照组的差异均具有统计学意义（P值均＜0.01）.结论 刺参黏多糖可能是通过Hippo信号通路及相关基因抑制胰腺癌SW1990细胞的增殖.

Objective To investigate the possible mechanism of inhibitory effect of stichopus japonicus acidic mucopolysaccharide （SJAMP） on the proliferation in pancreatic carcinoma SW1990 cells. Methods SW1990 cells were incubated with different concentrations of SJAMP for 24, 48, and 72 h. SW 1990 cells without SJAMP treatment were as control. RT-PCR was used to determine the expression of Hippo pathway members, including YAP, MST1 and related genes---Survivin, TEADI , Caspase-9, TEAD1 mRNA, and the protein expressions of YAP and phosphorylated YAP （p-YAP） in SW1990 cells were detected by Western blotting. Results The expression level in cells of control group was as 1, the YAP, TEAD1, Survivin, Caspase-9, MST1 mRNA expression after SJAMP treatment for 48 h were 0.27, 0.31, 0.26, 4.06, 7.06, and the expressions of YAP, TEAD1, Survivin mRNA were down-regulated, and the expressions of Caspase-9, MST1 mRNA were up-regulated in a time dependent manner, and the difference between the treatment group and control group was statistically significant （P 〈 0.05 or P 〈 0.01 ）. The expressions of YAP, pYAD protein in control group were 0. 705 ＋ 0.021, 0. 430 ＋ 0. 043, and after SJAMP treatment for 48 h, the expressions were 0. 545 ＋0.071, 0. 652 ＋0. 025, and the expression of YAD was down-regulated, and the expression of pYAD was up-regulated in a time dependent manner, and the difference between the treatment group and control group was statistically significant （P 〈 0. 01）. Conclutions SJAMP inhibits the proliferation of pancreatic carcinoma SW1990 cells possibly through Hippo pathway and relevant target genes.