扶正利湿抗癌方对LNCaP细胞中PSMA表达及p38 MAPK通路的影响

Effect of Fuzheng Lishi Kang’ai decoction on PSMA expression and p38 MAPK pathway in LNCaP cells

目的探讨扶正利湿抗癌方对人前列腺癌细胞(LNCaP)中前列腺特异性膜抗原(PSMA)及p38 MAPK通路的影响。方法将SD大鼠随机分为4组:对照组、低剂量组[12.5 g/(kg·d)]、中剂量组[25 g/(kg·d)]和高剂量组[50 g/(kg·d)],抽取各组含药血清,将其与培养的LNCaP细胞反应。利用18 F-PSMA1007探针检测LNCaP细胞中PSMA表达。采用蛋白质印迹法(Western blot)检测LNCaP细胞中p38 MAPK通路相关蛋白p38 MAPK、p-p38 MAPK、Bax、Caspase-3、P53及Bcl-2的表达。应用CCK-8法观察LNCaP细胞的增殖抑制情况。采用流式细胞仪分析LNCaP细胞凋亡情况。结果与对照组比较,低剂量组PSMA表达水平降低,p-p38 MAPK、Bax、Caspase-3及P53蛋白表达增高,Bcl-2蛋白表达降低,LNCaP细胞的增殖抑制率及细胞凋亡率升高(均P<0.05),而p38 MAPK蛋白表达差异无统计学意义(P>0.05)。与低剂量组比较,中剂量组PSMA表达水平降低,p-p38 MAPK、Bax、Caspase-3及P53蛋白表达增高,Bcl-2蛋白表达降低,LNCaP细胞的增殖抑制率及细胞凋亡率升高(均P<0.05),而p38 MAPK蛋白表达差异无统计学意义(P>0.05)。与中剂量组比较,高剂量组PSMA表达水平降低,p-p38 MAPK、Bax、Caspase-3及P53蛋白表达增高,Bcl-2蛋白表达降低,LNCaP细胞的增殖抑制率及细胞凋亡率升高(均P<0.05),而p38 MAPK蛋白表达差异无统计学意义(P>0.05)。结论扶正利湿抗癌方可能通过抑制PSMA表达,激活p38 MAPK信号传导通路从而抑制LNCaP细胞增殖,促进凋亡。

Objective To investigate the effects of Fuzheng Lishi Kang’ai decoction on the expression of prostate-specific membrane antigen(PSMA)and the p38 MAPK pathway in human prostate cancer LNCaP cells.Methods SD rats were randomly divided into four groups:control group,low-dose group[12.5 g/(kg·d)],medium-dose group[25 g/(kg·d)],and high-dose group[50 g/(kg·d)].Serum containing different concentrations of drug in each group was extracted and co-cultured with LNCaP cells.The expression of PSMA in LNCaP cells was detected using the 18 F-PSMA1007 probe.The expressions of p38 MAPK,p-p38 MAPK,Bax,Caspase-3,P53,and Bcl-2 proteins related to the p38 MAPK pathway in LNCaP cells were detected using Western blot.The proliferation inhibition of LNCaP cells was observed using the CCK-8 assay.The apoptosis of LNCaP cells was analyzed using flow cytometry.Results Compared with control group,the expression of PSMA was decreased in low-dose group,the expressions of p-p38 MAPK,Bax,Caspase-3,and P53 proteins were increased,the expression of Bcl-2 protein was decreased,the proliferation inhibition rate was increased,and the apoptosis rate of LNCaP cells was increased(all P<0.05),while the expression of p38 MAPK protein showed no significant difference(P>0.05).Compared with low-dose group,the expression of PSMA was decreased in medium-dose group,the expressions of p-p38 MAPK,Bax,Caspase-3,and P53 proteins were increased,the expression of Bcl-2 protein was decreased,the proliferation inhibition rate was increased,and the apoptosis rate of LNCaP cells was increased(all P<0.05),while the expression of p38 MAPK protein showed no significant difference(P>0.05).Compared with medium-dose group,the expression of PSMA was decreased in high-dose group,the expressions of p-p38 MAPK,Bax,Caspase-3,and P53 proteins were increased,the expression of Bcl-2 protein was decreased,the proliferation inhibition rate was increased,and the apoptosis rate of LNCaP cells was increased(all P<0.05),while the expression of p38 MAPK protein showed no significant difference(P>0.05).Conclusion Fuzheng Lishi Kang’ai decoction may suppress the proliferation and promote the apoptosis of LNCaP cells by inhibiting the expression of PSMA and activating the p38 MAPK signaling pathway.