硫化氢抑制前列腺癌骨转移PC-3细胞增殖和侵袭的体外研究

摘要

目的 探讨硫化氢对前列腺癌骨转移PC-3细胞增殖和侵袭的影响及其可能机制.方法 分别用0、1、2、4 mmol/LNaHS(硫化氢载体)作用前列腺癌骨转移PC-3细胞36 h,CCK-8比色法法检测细胞存活率,transwell法检测细胞侵袭能力,ELISA法检测细胞MMP-2和MMP-9分泌能力,Western-Blot法检测Bcl-2、Bax蛋白的表达.结果 硫化氢对PC-3细胞生长的影响:用硫化氢孵育PC-3细胞36 h,1.0 mmol/L NaHS组细胞存活率与对照组比较,差异无统计学意义(P＞0.05);2、4 mmol/L NaHS作用PC-3细胞36 h,细胞存活率降低,与对照组比较,能不同程度抑制PC-3细胞的生长,差异有统计学意义(P＜ 0.05或P＜0.01).硫化氢对PC-3细胞侵袭的影响:用硫化氢孵育PC-3细胞36 h,1 mmol/L NaHS组细胞侵袭数与对照组比较,差异无统计学意义(P＞0.05);2、4 mmol/L NaHS作用PC-3细胞36 h,细胞侵袭数减少,与对照组比较,能不同程度抑制PC-3细胞的侵袭,差异有统计学意义(P＜ 0.05或P＜0.01).ELISA法检测PC-3细胞MMP-2和MMP-9分泌能力:用硫化氢孵育PC-3细胞36 h,1 mmol/L NaHS组细胞MMP-2和MMP-9分泌量与对照组比较,差异无统计学意义(P＞0.05);2、4 mmol/L NaHS作用PC-3细胞36 h,MMP-2和MMP-9分泌量减少,与对照组比较,能不同程度抑制PC-3细胞MMP-2和MMP-9的分泌,差异有统计学意义(P＜ 0.05或P＜ 0.01).Western blot检测PC-3细胞Bcl-2/Bax蛋白表达:1、2、4 mmol/L NaHS组,Bcl-2表达降低,Bax表达升高,呈浓度依赖性.结论 2 mmol/L浓度以上的NaHS能影响Bcl-2和Bax表达和MMP-2 MMP-9分泌来抑制PC-3细胞的生长和侵袭.%Objective To investigate the effect of hydrogen sulfide on prostate cancer bone metastasis PC-3 cells proliferation and invasion in vitro and the possible mechanism.Methods PC-3 cells were treated with NaHS (a donor of hydrogen sulfide) at 0,1,2,4 mmol/L concentration for 36 h,respectively,then cell viability was tested by cell counter kit-8.Transwell assay were used to detect PC-3 cells invasive ability and ELISA was used to detect MMP-2and MMP-9 secretion.PC-3 cells were treated with NaHS at 0,1,2,4 mmol/L concentration for 36 h,then the expressions of Bcl-2,Bax were evaluated by Western-Blot assay.Results The effects of hydrogen sulfide on PC-3 cells proliferation:the differences of cell viabilitie were not significant (P ＞ 0.05) when PC-3 cells were treated with NaHS at 1 mmol/L concentrations.When PC-3 cells were treated with NaHS at 2,4 mmol/L concentration,the cell viability was declined,compare with control group (P ＜ 0.05,P ＜ 0.01).The effects of hydrogen sulfide on PC-3 cells invasive:the numbers of transfer to the other side of transwell membrane were counted in tumor invasive assay.The difference of 1 mmol/L NaHS group was not significant compared with control group (P ＞ 0.05) and 2 mmol/L NaHS group and 4mmol/L NaHS group were declined compared with control group (P ＜ 0.05 or P ＜ 0.01).The effect of hydrogen sulfide on MMP-2 and MMP-9 secretion:when cells were treated by 1 mmol/L NaHS,the secretion of MMP-2 and MMP-9was not significant compared with control group (P ＞ 0.05),2 mmol/L NaHS group and 4 mmol/L NaHS group were declined of MMP-2 and MMP-9 secretion compared with control group (P ＜ 0.05 or P ＜ 0.01).The effects of hydrogen sulfide on PC-3 cells Bcl-2 and Bax expression:the expression of Bcl-2 was declined and Bax expression increased compare with control group.Conclusion Hydrogen sulfide can inhibit the growth and invasive of PC-3 cells at the concentration 2,4 mmol/L by affecting Bcl-2 and Bax expression and reducing secretion of MMP-2 and MMP-9.