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目的:体外观察低剂量1,25-二羟基维生素D3[1,25-(OH)2D3,以下简称维生素D]预处理对多柔比星杀伤胃癌细胞株BGC-823的影响,并探讨其可能的作用机制。方法:MTT法检测维生素D对胃癌细胞株BGC-823的细胞毒作用,以无细胞毒时的浓度作为低剂量。MTT法测定低剂量的维生素D预处理后,多柔比星对BGC-823细胞的半数抑制浓度(half inhibitory concentration,IC50)的变化;FCM法检测低剂量维生素D预处理后,多柔比星致BGC-823细胞凋亡和死亡比率的变化;蛋白质印迹法检测低剂量维生素D处理对BGC-823细胞中Bax、Bcl-2、p38及磷酸化的p38蛋白表达的影响。结果:1nmol/L与10nmol/L维生素D对BGC-823细胞均无细胞毒作用(P>0.05),而10nmol/L维生素D预处理能降低多柔比星对BGC-823细胞的IC5(0P<0.05)。1nmol/L和10nmol/L维生素D预处理均能增强多柔比星对BGC-823细胞的杀伤作用(P<0.05)。1nmol/L与10nmol/L维生素D单独处理对BGC-823细胞中Bax、Bcl-2及p38蛋白表达水平均无明显影响(P>0.05),而10nmol/L维生素D能增强BGC-823细胞中磷酸化p38的表达(P<0.05)。结论:低剂量维生素D预处理能增强多柔比星对胃癌细胞的杀伤作用,其作用机制可能与提高细胞内p38蛋白的磷酸化水平有关。
OBJECTIVE: To observe the effect of low dose 1,25-dihydroxyvitamin D3 [1,25- (OH) 2D3, hereinafter referred to as vitamin D] on the gastric cancer cell line BGC-823 induced by doxorubicin The mechanism of action. Methods: The cytotoxic effect of vitamin D on gastric cancer cell line BGC-823 was determined by MTT assay. The concentration of cytotoxicity was taken as low dose. MTT assay was used to determine the change of half inhibitory concentration (IC50) of doxorubicin on BGC-823 cells pretreated with low dose of vitamin D. Pretreatment with low dose of vitamin D detected by doxorubicin 823 cells apoptosis and death rate changes; Western blotting detection of low-dose vitamin D treatment of BGC-823 cells Bax, Bcl-2, p38 and phosphorylation of p38 protein expression. Results: 1nmol / L and 10nmol / L vitamin D had no cytotoxic effect on BGC-823 cells (P> 0.05), while pretreatment with 10nmol / L vitamin D reduced the IC50 of doxorubicin in BGC- <0.05). Pretreatment with 1 nmol / L and 10 nmol / L vitamin D both enhanced the killing effect of doxorubicin on BGC-823 cells (P <0.05). Bcl-2 and p38 protein expression in BGC-823 cells was not significantly affected by 1nmol / L and 10nmol / L vitamin D alone (P> 0.05), while 10nmol / L vitamin D could enhance the expression of Bax, Bcl- Phosphorylated p38 expression (P <0.05). Conclusion: Low-dose vitamin D pretreatment can enhance the killing effect of doxorubicin on gastric cancer cells, and its mechanism may be related to the increase of intracellular phosphorylation of p38 protein.