神经酰胺激酶抑制剂NVP-231对去势抵抗性前列腺癌细胞恶性表型效应的研究

The effects of ceramide kinase inhibitor NVP-231 on the malignant phenotype of castration-resistant prostate cancer cells

目的探讨神经酰胺激酶(CerK)抑制剂NVP-231对去势抵抗性前列腺癌(CRPC)细胞增殖、凋亡、迁移和侵袭的影响。方法采用Western blotting法检测VCaP、LNCaP两株雄激素依赖性前列腺癌细胞与C4-2B、PC3两株CRPC细胞内CerK蛋白水平。用DMSO或不同浓度(500、1000 nmol/L)NVP-231分别处理C4-2B和PC3细胞。采用CCK-8法、克隆形成实验、Hoechst染色法、划痕愈合实验和Transwell小室实验分别检测处理48 h的细胞增殖、克隆形成、凋亡、迁移和侵袭能力。结果与VCaP、LNCaP两株雄激素依赖性前列腺癌细胞比较,C4-2B、PC3两株CRPC细胞中CerK蛋白水平明显升高,差异均有统计学意义(P<0.05)。与DMSO处理组比较,C4-2B和PC3细胞经NVP-231处理后细胞增殖能力降低,细胞克隆形成数目减少,细胞凋亡率升高,划痕愈合率降低,穿膜细胞数减少,差异均有统计学意义(P<0.05),且均具有浓度依赖性。结论CerK抑制剂NVP-231能通过抑制细胞增殖与生长,促进凋亡,降低细胞侵袭和迁移能力,进而逆转CRPC细胞的恶性表型。

Objective To investigate the effects of the ceramide kinase(CerK)inhibitor NVP-231 on cell proliferation,apoptosis,invasion and migration of castration resistant prostate cancer(CRPC)cells.Methods Western blotting was used to detect the CerK protein level in the human prostate cancer(PCa)cell lines,including CRPC cell lines(C4-2B and PC3)and androgen-dependent PCa cell lines(VCaP and LNCaP).C4-2B and PC3 were treated with NVP-231 at different concentrations(500,1000 nmol/L),respectively.CCK-8,clonogenesis test,Hoechst staining,scratch healing test and Transwell test were used to detect the cell proliferation,clonogenesis,apoptosis,migration and invasion after 48 h of treatment.Results Compared with VCaP and LNCaP androgen-dependent prostate cancer cells,the level of CerK protein in CRPC cells of C4-2B and PC3 increased significantly(P<0.05).Compared with DMSO treatment group,C4-2B and PC3 cells after the treatment of NVP-231,the proliferative capacity and the number of cell clones decreased,the rate of apoptosis increased,the rate of scratch healing and the number of penetrating cells were also decreased.The difference was statistically significant(P<0.05),and the results were concentration-dependent.Conclusion CerK inhibitor NVP-231 can reverse the malignant phenotype of CRPC cells by inhibiting cell proliferation and growth,promoting apoptosis,reducing cell invasion and migration.