预知子提取物增敏奥沙利铂诱导结肠癌HCT116细胞凋亡的研究

Study on Fructus Akebiae Extraction Inducing Apoptosis Sensitization of Colon Cancer HCT116 Cells to Oxaliplatin

    摘要
  • 摘要:
      目的  探讨预知子提取物增敏奥沙利铂(OXA)诱导结肠癌HCT116细胞凋亡的作用及机制, 为减少OXA毒副作用发生的临床应用提供研究基础。
      方法  将预知子乙酸乙酯萃取成分进行中压柱层析分离, 并进行抗肿瘤药效筛选, 获取活性部位B14, 并进一步分离和鉴定该活性部位成分; 运用MTT法检测B14、OXA及二者联用对HCT116的增殖抑制作用；采用CompuSyn软件评价联合药效, 确定药物联用最佳剂量; 流式细胞术检测B14与OXA联用后HCT116细胞的凋亡率及周期分布; Western blot法检测凋亡及信号通路相关蛋白的表达水平。
      结果  筛选确定预知子活性部分B14，检测并明确其主要成分为皂苷B和α-常春藤皂苷；6 μg·mL-1 B14可显著促进15 μg·mL-1 OXA对HCT116细胞的增殖抑制作用(P < 0.001)，促进细胞凋亡(P < 0.001)；联合组细胞周期被阻滞于G2/M期(P < 0.001)；Western blot结果显示，与OXA组比较，联合组Bcl-2、Caspase-3及Bcl-2/Bax比值显著降低(P < 0.01，P < 0.001)，Bax、Cyt-c、Cleaved caspase-3、Cleaved caspase-3/Caspase-3比值及p-p38 MAPK显著升高(P < 0.05，P < 0.01，P < 0.001)。
      结论  预知子提取物具有增敏OXA诱导HCT116凋亡的作用, 其机制可能与阻滞细胞周期在G2/M期及激活p38 MAPK介导的线粒体凋亡通路有关。研究结果为降低OXA毒副作用的临床应用提供了线索。

     

    Abstract:
      OBJECTIVE  To investigate the efficacy and mechanism of Fructus akebiae extraction (B14) inducing the apoptosis sensitization of colon cancer HCT116 cells to oxaliplatin (OXA), and provide the evidence for the prevention of side effect of OXA in clinical application.
      METHODS  The ethyl acetate extraction of Fructus akebiae were isolated by medium pressure column chromatography, and the active fraction B14 was obtained by anti-tumor experiment, further followed by research for its component identification. MTT assay was used to detect the proliferation inhibition of HCT116 by B14, OXA and their combination. CompuSyn software was used to evaluate the sensitization efficiency and determine the optimal dose of the combination. The apoptosis rate and cycle distribution of HCT116 cells were detected by flow cytometry. The expressions of apoptosis-related proteins were detected by Western blot.
      RESULTS  The main components of the active fraction B14 were identified as saponin B and α-hederin. 6 μg·mL-1 B14 combinated with 15 μg·mL-1 OXA significantly inhibited the proliferation and promoted the apoptosis of HCT116 cells (P < 0.001). The cell cycle of HCT116 was arrested at G2/M stage after the combination treatment (P < 0.001). Western blot results showed that, compared with OXA group, protein expressions of Bcl-2, Caspase-3 and the ratio of Bcl-2/Bax in combination group were significantly down-regulated (P < 0.01, P < 0.001), while Bax, Cyt-c, Cleaved caspase-3, the ratio of Cleaved caspase-3/Caspase-3 and p-p38 MAPK were significantly up-regulated (P < 0.05, P < 0.01, P < 0.001).
      CONCLUSION  The extraction of Fructus akebiae can induce the apoptosis sensitization of HCT116 cells to OXA, which may be related to the regulation of p38 MAPK pathway. This study may provide the evidence for preventing the side effect of OXA in clinical application.

     

    • HTML全文
    • 参考文献(22)
    • 相关文章
    • 施引文献
  • 资源附件(0)

/

返回文章
返回