苦参碱联合顺铂对人肾母细胞瘤SK-NEP-1细胞PDCD4表达的影响

Effect of matrine and cisplatin in combination on PDCD4 expression in SK-NEP-1 cells

摘要:

目的 探讨苦参碱对人肾母细胞瘤SK-NEP-1 细胞存活及凋亡的影响，及降低顺铂化疗耐药性的可能作用机制。方法 将SK-NEP-1 细胞分为对照组、5 μg/mL 顺铂组、苦参碱组（0.5、1.0 和1.5 mg/mL）、苦参碱（0.5、1.0 和1.5 mg/mL）联合5 μg/mL 顺铂组。应用MTT 比色法检测各组细胞存活率，流式细胞仪检测各组细胞凋亡率，RT-PCR 检测各组细胞的PDCD4 mRNA 表达。结果 各实验组SK-NEP-1 细胞的存活率均低于对照组（均P<0.01），苦参碱组随药物浓度增加，存活率逐渐降低，苦参碱联合顺铂组与单药苦参碱组比较，存活率均明显降低（均P<0.01）。各实验组SK-NEP-1 细胞的凋亡率均高于对照组（均P<0.05），苦参碱组随药物浓度增加，凋亡率逐渐增高，苦参碱联合顺铂组与单药苦参碱组比较，凋亡率均明显增高（均P<0.01）。各实验组PDCD4 mRNA 的表达水平均高于对照组（均P<0.01），苦参碱联合顺铂组与单药苦参碱组及顺铂组比较，PDCD4 mRNA 表达均显著提高（均P<0.01）。结论 苦参碱可浓度依赖性的地抑制SK-NEP-1 细胞的增殖并诱导凋亡，从而提高顺铂化疗的敏感性，该作用可能与提高细胞内PDCD4 mRNA 的表达有关。

Abstract:

Objective Matrine, a major ingredient of sophora, has an anti-tumor activity, capable of suppressing the proliferation and metastasis and promoting apoptosis or differentiation of tumor cells. This study was designed to investigate the effects of matrine on survival and apoptosis of nephroblastoma cell line SK-NEP-1, reduction of drugresistance of cisplatin and the mechanism(s) underlying these effects. Methods SK-NEP-1 cells were treated with matrine and cisplatin at various doses (0.5, 1.0 and 1.5 mg/mL), either each alone or in combination. The viability in treated SK-NEP-1 cells was assessed by MTT colorimetric assay, apoptosis by flow cytometry, and PDCD4 mRNA abundance by RT-PCR. Results As compared with the non-treatment control, matrine and cisplation, regardless of combination and dosage, significantly reduced the viability (P<0.01), induced apoptosis (P<0.01), and increased PDCD4 mRNA abundance (P<0.01), in SK-NEP-1 cells. The above effects of matrine and cisplation were dose-dependent when they were used alone, and were more pronounced when they were used in combination (P<0.05). Conclusions Matrine can significantly induce apoptosis and inhibit growth of SK-NEP-1 cells in a dose-dependent manner, thus increasing the chemotherapeutic sensibility of cisplatin. The observed effects of matrine may be a result of increased PDCD4 expressi on.