狼毒大戟提取液治疗Lewis肺癌机制的探讨

Research on the optimize treating mechanism of LDE for Lewis lung cancer

目的:探讨狼毒大戟提取液优化治疗非小细胞肺癌的机制。方法:收集狼毒干预24h后的细胞,流式细胞技术分析肿瘤细胞周期及凋亡率;real time PCR检测Caspase-9mRNA的表达。收集对数生长期细胞接种至C57BL/6小鼠右侧腋窝下,随机分组,腹腔给药后计算抑瘤率;流式细胞技术分析肿瘤细胞周期及凋亡率;real time PCR检测Bcl-2mRNA和Bax mRNA的表达,抗氧化能力检测法检测CAT和SOD的表达。结果:体内外实验显示,狼毒大戟能明显促进肿瘤细胞的凋亡,将细胞周期阻滞在S期,并能明显促进Bax mRNA、抑制Bcl-2mRNA的表达;体内实验也显示狼毒大戟可以明显提高机体抗氧化能力。结论:狼毒大戟在促进肿瘤细胞凋亡的同时可以提高机体抗氧化能力,充分体现了狼毒大戟优化治疗肿瘤的优势,前景广阔,值得进一步研究,为临床应用提供依据。

OBJECTIVE： To explore the mechanism of Lang-du extrct（LDE） for Lewis lung cancer. METHODS： By technique of cell culture in vitro, Lewis cells were treated with LDE in different concentrations for 24 hours, then apoptotic rate and cell cycle were measured by flow cytometer; The mRNA expression of Bcl-2 and Bax were assessed by Realtime PCR. C57BL/6 mices were transplanted subcutaneously to the right limb of 1 × 106 Lewis cells per mice. After transplan- ted,the mice were separaed into 2 groups：control and LDE group. The LDE group were given LDE intraperitoneal injec- tion. The two groups of mice were killed after 7 days, then, tumor were taken off, they were weighed firstly for mesure ment of inhibition rate,apotosis rate,and cell cycle were analyzed with flow cytometry;Bcl-2 and Bax mRNA were detec- ted by Realtime PCR; Some indexes about lipid peroxidation such as SOD,GSH-Px and CAT were determined. RESULTS： In vitro and in vivo experements indicated that the LDE could significantly induce apoptosis and block the cell cycle at Go / G1 phase; Realtime pcr showed that Bax expression level was significantly increased and Bcl-2 reduced; Comepared to control group, the result shows statistical difference in the CAT, SOD, GSH-Px value. Inclusion：LDE could effectively in- duce apoptosis of Lewis cells through blocking the cell cycle at Go/G1 phase. CONCLUSIONS： The increase of Caspase 9 expression level may play an important role in inducing Lewis cell apotosis. LDE can induce the apoptosis of Lewis cells and enhance the Antioxidant ability of mouse.