蛞蝓提取物抗肺癌的作用机制研究

目的探讨蛞蝓提取物(EL-3)抗肺癌的作用机制。方法 (1)溴化尿嘧啶核苷(BrdU)掺入法观察EL-3对人源性肺癌A549细胞的掺入抑制率;(2)吖啶橙/溴化乙啶(AO/EB)染色荧光显微镜,观察EL-3诱发A549凋亡细胞的形态,计数凋亡细胞;PI染色流式细胞术(FCM)分析细胞周期和凋亡率。(3)DNA琼脂糖凝胶电泳法,观察凋亡细胞"梯形"DNA条带;(4)免疫组化DAB染色法观察EL-3对PCNA、VEGF、CD31的蛋白表达的影响。结果 (1)单用蛞蝓提取物100μg/mL浓度的EL-3对人肺癌细胞A549核酸合成抑制率(IR)高达63.59%;与空白对照组(0)和溶媒对照组(7.54%)比较差异有统计学意义(P<0.01),且呈剂量依赖性,其半数抑制浓度为28.53μg/mL;(2)与化疗药合用有增效作用:EL-3与顺铂合用能显著提高顺铂对A549细胞的核酸合成抑制率:EL-3低、中、高剂量组,使单用DDP的IR由14.01%分别提高到44.5%、48.96%和67.05%(P<0.01);IC50为20.62μg/mL;EL-3与紫杉醇合用能显著提高紫杉醇对A549细胞的核酸合成抑制率。EL-3低、中、高剂量组使单用TAX对A549细胞的IR由21.79%,分别提高到58.35%、66.13%和73.67%,差异有统计学意义(P<0.05),IC50值为7.41μg/mL;(3)EL-3能有效地诱导A549细胞凋亡;(4)EL-3具有抑制裸鼠移植瘤细胞PCNA和VEGF及CD31蛋白表达作用。结论 EL-3抗肺癌的作用机制是多靶点复合性的有:(1)抑制肺癌细胞核酸合成;(2)诱导肺癌细胞凋亡;(3)抑制癌灶的血管生成和癌细胞对血管壁的黏附。

Research of EL-3 on anti-lung cancer mechanism

Objective To investigate the mechanism of extract of limax(EL-3) on anti-lung cancer.Methods(1) BrdU infiltration method was used to observe the inhibiting rates of EL-3 on the human lung cancer A549 cells.(2) The morphology of A549 cells apoptosis induced by EL-3 was determined and counted with the acridine orange/ethidium bromide(AO/EB) staining,and the cell cycle and apoptosis were analyzed with FCM.(3) The ladder-like DNA bands of apoptosis cells was observed by the DNA garose gelatin electrophoresis.(4...