苦参碱对人横纹肌肉瘤RD细胞增殖和凋亡的影响

Effects of matrine on the proliferation and apoptosis of human rhabdomyosarcoma RD cells

摘要:

目的：探讨苦参碱（matrine, Mat）对体外人横纹肌肉瘤RD细胞增殖和凋亡的影响，以及Mat诱导RD细胞凋亡的相关机制。方法：采用MTT比色法检测终浓度为0.5、1.0、1.5和2.0 mg/mL Mat对RD细胞的增殖抑制率；流式细胞术检测上述4种不同浓度Mat对RD细胞的凋亡作用；RT-PCR法检测终浓度为0.5、1.0和1.5 mg/mL Mat对RD细胞Cyclin D1及Survivin mRNA表达的影响。结果：各浓度实验组RD细胞的增殖抑制率和凋亡率均高于未经Mat处理的对照组（均P<0.01），且随着药物浓度增加，细胞增殖抑制率逐渐增高，呈剂量依赖性。RT-PCR检测Cyclin D1 及Survivin mRNA在各组RD细胞中均有表达，两者的表达水平在各浓度处理组中与对照组相比均有明显下降（P＜0.01）。其中Cyclin D1在各浓度组间比较差异均有统计学意义（P＜0.05），而Survivin仅在0.5 mg/mL 和1.5 mg/mL组间差异有统计学意义（P＜0.05）。结论：Mat能明显抑制RD细胞的增殖，并诱导其凋亡；其抗肿瘤机制可能与下调Survivin和Cyclin D1 mRNA的表达有关。

Abstract:

OBJECTIVE: To investigate the effects of matrine on the proliferation and apoptosis of human rhabdomyosarcoma RD cells in vitro, and to explore the mechanism of matrine inducing apoptosis of RD cells. METHODS: MTT assay was used to measure the proliferation inhibition rates of RD cells that were treated with matrine (final concentrations= 0.5, 1.0, 1.5, and 2.0 mg/mL). Flow cytometry was used to evaluate the apoptosis of RD cells treated with the four concentrations of matrine. RT-PCR was used to measure the mRNA expression of cyclin D1 and survivin in RD cells treated with 0.5, 1.0, and 1.5 mg/mL of matrine. RESULTS: The RD cells treated with various concentrations of matrine showed significantly higher proliferation inhibition rates and apoptotic rates than those that were not treated with matrine (P<0.01), and with increased matrine concentration, the proliferation inhibition rate of RD cells increased gradually, thus exhibiting a dose dependence. The mRNA expression of cyclin D1 and survivin was seen in all RD cells, but was significantly lower in RD cells treated with matrine than in those that were not treated with matrine (P<0.01). There were significant differences in cyclin D1 mRNA level among the RD cells treated with 0.5, 1.0, and 1.5 mg/mL of matrine (P<0.05), while there was significant difference in survivin mRNA level between the RD cells treated with 0.5 and 1.5 mg/mL of matrine (P<0.05). CONCLUSIONS: Matrine can significantly inhibit proliferation and induce apoptosis of RD cells, which may be related to downregulating the mRNA expression of cyclin D1 and survivin.