前列腺特异性膜抗原通过p38通路对前列腺癌细胞增殖、迁移的调控研究

Regulatory effects of prostate specific membrane antigen on proliferatation, survival and metabolism of prostate cancer cells via p38 pathway

目的 观察前列腺特异性膜抗原（PSMA）在前列腺癌细胞增殖、迁移和分裂代谢过程中调控相关通路的作用.方法 实验对象包括稳定阻断PSMA表达的细胞株（干扰组）、未阻断PSMA表达的LNCaP细胞株（非干扰组）、不作任何处理的LNCaP细胞株（空白组）.利用Western blot及免疫荧光观察3组细胞磷酸化p38（p-p38）的表达量,并使用细胞计数试剂盒（CCK-8）法检测细胞增殖能力,Transwell小室检测细胞迁移能力,流式细胞仪检测细胞周期.结果 Western blot及细胞免疫荧光提示干扰PSMA表达后,p-p38表达水平下降40％（P＜0.05）;在SB203582（p38抑制剂）作用下,3组细胞p-p38均处于较低水平,组间比较差异无统计学意义（P＞0.05）.CCK-8法、Transwell法、流式细胞仪检测细胞周期提示PSMA干扰后细胞增殖、迁移能力下降,细胞S期百分比降低;给予SB203582后,3组细胞增殖、迁移能力明显下降,细胞S期百分比均处于低水平.结论 PSMA通过上调p38通路对细胞的增殖、细胞周期产生影响,从而对LNCaP细胞起正性调节作用.

Objective To test the role of prostate specific membrane antigen （PSMA） regulating related pathways in the proliferatation,survival and metabolism of prostate cancer cells.Methods LNCaP cells had been stably transfected with lentivirus-mediated shRNA for PSMA silencing in previous study.The efficacy of PSMA knockdown was testified in LNCaP cell line.By using this PSMA-LNCaP cell line,the expression of PSMA and phospho-p38 （p-p38） detected by using Western blotting was compared among groups.Immunofluorescence was used to confirm the change of p-p38 in cells.Cell viability and migration were measured by cell counting kit-8 reagent and Transwell analysis respectively.Flow cytometry was employed to evaluate the cell survival.P38 pathway inhibitor （SB203582） was used in the culture medium to determine the role of p38 in the prostate cancer cells regulated by PSMA.Results After silencing the expression of PSMA,the expression level of p-p38 was decreased by approximate 40％ as compared with the blank and non-interference groups （P ＜ 0.05）.When the cells were incubated with SB203582,the p-p38 expression in three groups was at a low level and no significant difference was found among groups （P ＞0.05）.The results of immunofluorescence further proved the relationship between PSMA and p-p38.Decrease of cell viability,migration and survival was observed upon PSMA silencing.SB203580,a specific inhibitor of p38 mitogen-activated protein kinase （MAPK） pathway,also reduced the proliferation,migration and survival of LNCaP cells.Conclusion These data suggest PSMA may stimulate proliferation,migration and survival of prostate cancer cells through p38 MAPK pathway,revealing a novel mechanism for PSMA playing positive roles in LNCaP cells.