TRPM8对前列腺癌细胞PC-3细胞增殖和迁移影响的研究

目的探讨TRPM8对雄激素非依赖性前列腺癌PC-3细胞增殖和迁移能力的影响。方法PC-3细胞分别转染空载体pcDNA3和目的基因TRPM8，筛选稳定表达细胞株；分别通过免疫荧光和钙成像检测TRPM8蛋白表达、分布以及功能；通过流式细胞术和划痕实验检测TRPM8对PC-3细胞周期和细胞迁移率的影响。结果免疫细胞化学和钙成像提示PC-3-TRPM8细胞表达有功能的TRPM8通道；TRPM8能诱导细胞周期阻滞于G0／G1期，与PC3和PC-3-vector细胞相比，处于G0／G1期的PC-3-TRPM8细胞显著增加（71．89％±3．43％vs54．67％±4．59％、51．48％±3．54％，P〈0．05），易化饥饿诱导的细胞凋亡（12．56％±1．78％vs4．67％±1．49％、5．28±1．35％，P〈0．05），并抑制细胞迁移（P〈0．01）。结论TRPM8并不为PC-3细胞存活所必需，相反，高表达的TRPM8能抑制PC-3细胞的增殖和迁移，可能为前列腺癌治疗的一个新靶点。

Effects of TRPM8 on the proliferation and motility of prostate cancer PC-3 cells

Objective To investigate the effects of transient receptor potenlial M8 （TRPM8） channel on the proliferation and motility of androgen-independent prostate cancer PC-3 cells. Methods After being permanently transfected with an empty vector and cDNA encoding the TRPM8 pro tein,cells were analysed for cell cycle distribution and motility using flow cytometry and scratch assay. Results Immunocytochemistry and Ca^2＋ imaging analysis revealed the overexpression of func tional TRPM8 channel on both endoplasmie reticulum and plasma membrane of PC-3-TRPM8 cells. Cell cycle distribution and scratch assay analysis revealed that TRPM8 induced cell cycle arrest at the G0/G1 stage （ 71.89% 3.43% vs 54.67%±4.59%,51.48%±3.54% ,P〈0. 05 ） and facilitated the cell apoptosis induced by starvation （ 12. 56% ± 1. 78% vs 4.67%
± 1. 49%, 5.28 ±1.35% ,P〈0.05 ）. Furthermore,TRPM8 inhibited the migration of PC 3 TRPM8 cells （P〈0.01） through the inactivation of focal adhesion kinase. Conclusions It appears that TRPM8 was not essential for the survival of PC -3 cells;however,the overexpression of TRPM8 had negative effects on the proliferation and migration of PC- 3 cells. Thus,TRPM8 and its agonists may serve as important targets for the treatment of prostate cancer.