注射用磷酸肌酸钠对体外培养人卵巢癌细胞SKOV3侵袭能力的影响及其机制

Effect of creatine phosphate sodium for injection on invasion of human ovarian carcinoma SKOV3 cells in vitro and its mechanism

目的:观察注射用磷酸肌酸钠(CP)对人卵巢癌细胞SKOV3体外侵袭能力的影响,并探讨其作用机制。方法:体外培养人卵巢癌细胞SKOV3,随机分为空白对照组(生理盐水)及药物处理组(1、6和12 mmol.L-1CP);分别用生理盐水、CP处理细胞72 h后,采用MTT比色法和Transwell小室法观察细胞黏附和迁移侵袭能力;采用RT-PCR和FCM测定nm23-h1、c-myc mRNA及蛋白表达和MMP-2、MMP-9蛋白表达水平。结果:与空白对照组比较,1 mmol.L-1CP组各项指标差异无统计学意义(P>0.05);6和12 mmol.L-1CP组细胞黏附、迁移侵袭抑制率明显增加(P<0.01),nm23-h1 mRNA及蛋白表达水平明显升高(P<0.01),c-myc mRNA及蛋白表达水平明显降低(P<0.01);但6与12 mmol.L-1CP组间比较差异无统计学意义(P>0.05)。与空白对照组比较,1、6和12 mmol.L-1CP组MMP-2、MMP-9蛋白表达水平差异无统计学意义(P>0.05)。结论:一定浓度注射用CP可抑制体外培养SKOV3细胞的黏附和迁移侵袭能力,其机制可能与上调nm23-h1、下调c-myc基因表达有关。

Objective To observe the effect of creatine phosphate sodium（CP） on the invasion of human ovarian carcinoma SKOV3 cells in vitro and explore its mechanism.Methods The SKOV3 cells cultivated in vitro were treated with normal saline（control group）,1,6 and 12 mmol·L-1CP for 72 h,respectively.The adhesive ability of cells in each group was detected by MTT.The invasive and migrative abilities of cells in each group were detected by Transwell method.The expressions of nm23-h1,c-myc,MMP-2 and MMP-9 were analyzed with RT-PCR and FCM.Results Compared with control group,the indicators mentioned above in 1 mmol·L-1 CP grooup had no significantly changes（P0.05）;and the invasive and migrative abilities in 6 and 12 mmol·L-1CP groups were significantly decreased（P0.05）.The RT-PCR and FCM results showed that the nm23-h1 gene and protein expressions in 6 and 12 mmol·L-1CP groups were significantly increased compared with control group（P0.05）;but there was no significant difference between 6 mmol·L-1CP group and 12 mmol·L-1 CP group（P0.05）.Compared with control group,the expressions of c-myc in 6 and 12 mmol·L-1CP groups were significantly decreased（P0.05）,but there was no significant difference between 6 mmol·L-1CP group and 12 mmol·L-1CP group（P0.05）.The MMP-2 and MMP-9 protein expressions in 1,6 and 12 mmol·L-1CP groups had no significant changes compared with control group（P0.05）.Conclusion CP can significantly inhibit the adhesion,invasion,migration of SKOV3 cells and its mechanism may be related to up-regulation of the expression of nm23-h1 and down-regulation of the expression of c-myc.