环磷酰胺和吡柔比星对横纹肌肉瘤RD细胞株CXCR4表达的影响

Effects of Pirarubicin and Cyclophosphamide on expression of CXCR4 in human rhabdomyosarcoma RD cells and its sensitivity to chemotherapy

目的观察敏感剂量下环磷酰胺(CTX)和吡柔比星(THP)对体外培养人横纹肌肉瘤RD细胞株CXCR4基因表达的影响。方法体外培养RD细胞株;MTT试验明确CTX和THP对RD细胞的效应剂量;细胞划痕实验检测RD细胞的迁移能力;RT-PCR及Western blot法检测RD细胞CXCR4表达。结果 (1)敏感剂量下CTX(30 mmol/L)和THP(1000 ng/mL)均可抑制RD细胞的迁移(P<0.05),两药联合应用对细胞迁移抑制作用增强,较对照组差异有非常显著性(P<0.01);(2)各药物处理组(CTX,THP,CTX+THP)药物作用于RD细胞24 h后CXCR4蛋白的表达较对照组减少,其差异均有显著性(P<0.05);其中CTX+THP组较CTX组CXCR4蛋白表达减少更明显(P<0.05),但与THP组比较,差异无显著性(P>0.05);(3)同样条件下,各药物处理组(CTX,THP,CTX+THP)与对照组比较,RD细胞CXCR4 mRNA的表达均减少,差异均有显著性(P<0.05);CTX+THP组较CTX组CXCR4 mRNA表达减少明显(P<0.05),但较THP组差异无显著性(P>0.05)。结论化疗药物CTX和THP均能抑制人横纹肌肉瘤RD细胞的增殖;敏感剂量下的两种药物均能抑制RD细胞的迁移及下调RD细胞CXCR4的表达,提示两种化疗药物发挥作用的机制可能涉及到CXCR4基因。

[ Abstract] Objective To investigate the expression of chernokine CXC motif receptor 4 CXCR4 in human rhabdomyosarcoma RD cells, and its sensitivity to chemotherapeutic drugs CTX and THP, and to explore this chemokine receptor＇ s mechanism under sensitive dose about CTX/THP. Methods Human rhabdomyosarcoma RD cells were cultured in vitro. Dose-effects of chemotherapeutic drugs including CTX, THP on human rhabdomyosarcoma RD cells were investigated by MTT assay. The migratory ability was examined by wound healing assay. Expression of protein and mRNA of CXCR4 human rhabdomyosarcoma RD cells were detected by Western blotting and RT-PCR. Results CXCR4 expressed in human rhabdomyosarcoma RD cells. Dose-effects of chemotherapeutic drugs including CTX, THP are 30 mmol/L and 1000 ng/mL. Dose-effects of chemotherapeutic drugs including CTX, THP could inhibit rhabdomyosarcoma RD cells migration in vitro （ P 〈 0.05 ）, synergistic effect （ P 〈 0.01 ） was found between two medicines. Chemotherapeutic drugs CTX, THP decreased （ P 〈 0. 05 ） the expression of CXCR4 protein and mRNA level in human rhabdomyosarcoma RD cells, but no synergistic effect （ P 〉 0. 05 ） was found between two medicines. Conclusions CTX and THP could inhibit human rhabdomyosarcoma RD cells migration in vitro and down-regulate the expression of chemokine receptor CXCR4, indicating that chemotherapy can inhibit the progression of tumor cells by down-regulating the expression of CXCR4.