【作者】 李野；

【导师】 辛毅；

【作者基本信息】 大连医科大学 ， 生物化学与分子生物学， 2009， 硕士

【摘要】 恶性肿瘤是严重威胁人类生命安全的疾病之一。寻求高效低毒的抗肿瘤药物及治疗方法一直是科学界研究的热点。蝎毒作为一种生物毒素,凭借其广泛的生理学效应,极具研究价值。本文旨在研究蝎毒对肿瘤细胞的杀伤作用和敏感细胞系的筛选,并对其作用机制进行探讨,为蝎毒作为抗肿瘤药物的开发及临床应用提供实验依据。目的:观察蝎毒粗提物对人乳腺癌细胞MCF-7的抑制作用,初步探讨其作用机制。方法:采用MTT法和荧光免疫技术在三种常见肿瘤细胞(人肝癌细胞SMMC-7721、人乳腺癌细胞MCF-7和人宫颈癌细胞Hela)中选出对蝎毒敏感性较高的肿瘤细胞。免疫细胞化学法检测药物干预后,两种相关凋亡蛋白Caspase-3和Bcl-2在蛋白水平的表达量。流式细胞仪观察药物对细胞周期的影响,Western Blotting检测药物干预后,细胞周期相关蛋白CyclinD1在蛋白水平表达的变化。结果:MTT结果显示600μg/ml蝎毒粗提物对7721和MCF-7细胞均有抑制作用,与阴性对照组相比p<0.05.免疫荧光技术显示7721细胞对蝎毒的敏感性好于Hela细胞。免疫细胞化学检测显示药物干预后,MCF-7细胞中Caspase-3表达上调,Bcl-2表达下调。流式细胞仪显示药物干预后,处于S期细胞数减少,G0/G1期细胞数增多,与阴性对照组相比p<0.05。Western Blotting显示细胞周期相关蛋白CyclinD1蛋白表达量减少。结论:蝎毒粗提物可抑制MCF-7细胞的生长和增殖,其作用机制可能与诱导细胞发生凋亡,影响细胞G0/G1期和S期有关,在乳腺癌治疗中具有重要价值。更多还原

【Abstract】 Cancer is a serious threat to human life and safety of one of the diseases. To seek efficient and low toxicity of anticancer drμgs and treatment of the scientific community has been on the hot spots. Scorpion venom is a biological toxin, because of its wide range of physiological effects closely investigated. This paper aims to study the destruction about scorpion venom on human breast cancer cell line MCF-7 and sensitive cell line, and their mechanisms were discussed for the development of scorpion toxin of anti-tumor clinical applications experimental basis.Objective: detected growth inhibition of using the extractive of scorpion venom on human breast cancer cell line MCF-7, and investigated the mechanism.Methods: bolting the sensitive cell line in hepatoma cell SMMC-7721、Hela cell and breast cancer cell MCF-7 by MTT and IMF; detecting the apoptosis-related proteins expressed levels of caspase-3 and Bcl-2 proteins with ICC; observed the effect on cell cycle by FCM; detecting the cell cycle-related protein expressed levels of cyclinD1 with western blotting. Results: MTT method displayed that the inhibition on 7721 and MCF-7 by using the 600μg/ml extractive of scorpion venom, compared with the negative group(p<0.05);IMF method displayed that the sensitive to scorpion venom of 7721 cell was better than Hela cell after drug intervention; ICC method displayed that apoptosis protein caspase-3 increased and anti-apoptosis protein Bcl-2 decreased after drug intervention; FCM displayed that the amount of S period of time decreased, the amount of G1 period time increased after drug intervention, compared with the negative group(p<0.05); western blotting displayed that the expression of cell cycle-related protein cyclinD1 decreased after drug intervention.Conclusion: the extractive of scorpion venom inhibited the growth of human breast cancer MCF-7, and induced apoptosis in MCF-7 cell, the mechanism maybe related to G1 and S period of time, there was a significant value in treating human breast cancer.更多还原