斑蝥酸钠联合多西紫杉醇诱导A549细胞凋亡及对Caspase-8 表达的影响

Research on the Combined Effect of Sodium Cantharidinate and Docetaxel on Adenocarcinoma Cell Apoptosis and the Activities of Caspase-8

关键词:  斑蝥酸钠  多西紫杉醇  人肺腺癌A₅₄₉细胞  细胞凋亡  Caspase8

Key words:  sodium cantharidinate  docetaxel  A₅₄₉ cell  apoptosis  caspase8

摘要:

目的 探讨斑蝥酸钠联合多西紫杉醇抑制人肺腺癌A₅₄₉细胞增殖、诱导细胞凋亡的作用，初步探讨其作用机制。方法 采用MTT法测定细胞增殖抑制率，Annexin V-FITC/PI染色，流式细胞仪测定细胞凋亡率；Western印迹法测定Caspase-8活性。结果 斑蝥酸钠和多西紫杉醇均能抑制人肺腺癌A₅₄₉细胞的增殖(P<0.05)，合用与单用相比抑制率明显提高，诱导12 h时CI值为0.88，24 h时为0.96，36 h时为1.02。Annexin V-FITC/PI染色流式细胞仪检测提示斑蝥酸钠可增强多西紫杉醇的凋亡诱导作用。Western印迹法显示多西紫杉醇、斑蝥酸钠及联合加药组均可见凋亡相关蛋白Caspase-8的裂解条带，联合加药组的裂解条带最明显。结论 多西紫杉醇和斑蝥酸钠均可明显抑制人肺腺A549细胞增殖诱导细胞凋亡，联合使用作用更强，多西紫杉醇和斑蝥酸钠诱导细胞凋亡可能与促进Caspases-8表达有关。

Abstract:

OBJECTIVE To investigate the inhibition effects of combination of Docetaxel and Sodium Cantharidinate on human pulmonary adenocarcinoma. METHODS Adenocarcinoma cell line A₅₄₉was chosen and growth inhibition was measured both with MTT colorimetry and FCM assay. Caspase8 was detected by Western blot. RESULTS Either Docetaxel or Sodium Cantharidinate can inhibit the proliferation of A₅₄₉ cell respectively, however, Sodium Cantharidinate can increase the sensitivity of A₅₄₉ cell to Docetaxel when using them at the same time. The difference between single use and combine two drugs is distinct(P<0.05). High-level Caspase8 expression was detected with Docetaxel or Sodium Cantharidinate in A₅₄₉cells by Western blot. CONCLUSION Sodium Cantharidinate can sensitize A₅₄₉ cell to Docetaxel's effects in vitro. The induction of apoptosis may be related to expression of Caspase8.