论文部分内容阅读
目的探讨神经鞘鞍醇激酶1(Sphk1)抑制剂SKI-Ⅱ增敏顺铂(DDP)对SGC7901/DDP细胞周期的阻滞作用,并探讨其可能作用机制。方法将人胃腺癌耐DDP细胞株SGC7901/DDP进行体外培养后,分对照组、DDP组(DDP 2.50 mg/L)、SKI-Ⅱa组(SKI-Ⅱ1.25μmoL/L)、SKI-Ⅱb组(SKI-Ⅱ10.00μmoL/L)、SKI-Ⅱc组(SKI-Ⅱ1.25μmoL/L+DDP 2.50 mg/L)、SKI-Ⅱd组(SKI-Ⅱ10.00μmoL/L+DDP 2.50 mg/L)进行实验。采用MTT、流式细胞技术检测各组细胞生长、周期阻滞情况;免疫组化染色、Western blot法检测细胞Sphk1、P-gp、p27蛋白表达;并分析Sphk1与P-gp、p27蛋白的相关性。结果 DDP组、SKI-Ⅱa组对细胞生长、细胞周期无影响;与对照组比较,SKI-Ⅱc组可抑制细胞生长,使停滞于G0/G1的细胞增加、S期细胞降低(P<0.05)。与对照组比较,DDP组的Sphk1、P-gp、p27蛋白阳性表达率无统计学差异,SKI-Ⅱa组p27蛋白阳性表达率升高(P<0.05)。Pearson相关分析显示,Sphk1与P-gp呈正相关(r=0.550,P<0.01),与p27呈负相关(r=-0.663,P<0.01)。Western blot检测显示,DDP组未见明显改变,SKI-Ⅱb组、SKI-Ⅱd组的Sphk1、P-gp表达减弱,p27表达增强。结论 SKI-Ⅱ可通过抑制Sphk1蛋白下调P-gp蛋白、上调p27蛋白表达,增敏DDP对SGC7901/DDP细胞的周期阻滞作用。
Objective To investigate the blockade of the cell cycle of SGC7901/DDP by sensitized cisplatin (DDP) with SKI-II inhibitor Spik1, and to explore its possible mechanism. Methods Human gastric adenocarcinoma-resistant DDP cell line SGC7901/DDP was cultured in vitro and divided into control group, DDP group (DDP 2.50 mg/L), SKI-IIa group (SKI-II 1.25 μmoL/L), SKI-IIb group ( SKI-II 10.00 μmol/L), SKI-IIc group (SKI-II 1.25 μmol/L+DDP 2.50 mg/L), SKI-IId group (SKI-II 10.00 μmol/L+DDP 2.50 mg/L) . MTT and flow cytometry were used to detect cell growth and cell cycle arrest. Immunohistochemical staining and Western blot were used to detect the expression of Sphk1, P-gp and p27 proteins. The correlation between Sphk1 and P-gp and p27 protein was analyzed. Sex. Results The DDP group and SKI-IIa group had no effect on cell growth and cell cycle. Compared with the control group, SKI-IIc group could inhibit cell growth, increased the number of cells arrested in G0/G1 and decreased in S phase (P<0.05). . Compared with the control group, the positive rates of Sphk1, P-gp, and p27 proteins in the DDP group were not statistically different, and the positive rate of p27 protein in the SKI-IIa group was higher (P<0.05). Pearson correlation analysis showed that Sphk1 was positively correlated with P-gp (r=0.550, P<0.01) and negatively correlated with p27 (r=-0.663, P<0.01). Western blot analysis showed no significant changes in the DDP group. The expression of Sphk1 and P-gp in the SKI-IIb group and SKI-IId group was weakened, and p27 expression was enhanced. Conclusion SKI-II can attenuate the cycle arrest effect of DDP on SGC7901/DDP cells by inhibiting Sphk1 protein down-regulation of P-gp protein and up-regulating p27 protein expression.