论文部分内容阅读
目的:肿瘤是威胁人类生命安全的疾病之一。寻求高效的抗肿瘤药物一直是研究的热点。研究表明,蟾酥具有抗肿瘤等多种活性。本研究通过对体外培养的Hep-2细胞系进行药物干预,研究蟾酥对肿瘤细胞生长的抑制作用及其对Survivin蛋白和Caspase-9蛋白表达的影响,探讨蟾酥抗肿瘤的分子机制,为药物的临床应用提供新的理论基础和实验依据。方法:Hep-2细胞的传代培养。MTT法分别检测不同时间(6、12、24小时)、药物浓度(18、9、4.5μg/ml)的生长抑制率。Western blot检测4.5μg/ml浓度药物作用不同时间(0、6、12、24、48小时)Survivin和Caspase-9蛋白表达的影响。结果:蟾酥对Hep-2细胞的生长具有抑制作用。其抑制程度与蟾酥水溶物浓度高低和作用时间成正比,组间有显著差异(P<0.05)。Western blot结果显示:与对照组相比,受到蟾酥水溶物作用的Hep-2细胞系的Survivin与Caspase-9蛋白的表达呈现下降趋势,具有时间依赖性(P<0.05),统计学上具有显著差异。结论:Western Blot结果显示由于蟾酥水溶物的影响,Hep-2细胞的Survivin蛋白的表达减少、Caspase-9蛋白的表达减少。MTT检测结果显示随着药物浓度的升高、时间的延长,蟾酥水溶物对Hep-2细胞的生长抑制效果增强。
Objective: Tumor is one of the diseases threatening human life safety. Seeking efficient anticancer drugs has always been a research hotspot. Studies have shown that toad has a variety of anti-tumor activity. In this study, Hep-2 cells cultured in vitro drug intervention to study the inhibitory effect of toad on the growth of tumor cells and its impact on Survivin protein and Caspase-9 protein expression, explore the molecular mechanism of toad anti-tumor drug for the Clinical application of new theoretical basis and experimental basis. Methods: Hep-2 cells were subcultured. MTT assay were detected at different times (6,12,24 hours), drug concentration (18,9,4.5 g / ml) growth inhibition rate. Western blot was used to detect the effect of 4.5μg / ml drug on the expression of Survivin and Caspase-9 at different times (0, 6, 12, 24, 48 hours). Results: Toad could inhibit the growth of Hep-2 cells. The inhibition was proportional to the concentration of toad water soluble substance and the action time, there was a significant difference between the two groups (P <0.05). The results of Western blot showed that the expression of Survivin and Caspase-9 in Hep-2 cell line treated with water toad water had a decreasing trend compared with that in control group, with a time-dependent (P <0.05) statistically significant difference. Conclusion: The results of Western Blot showed that the expression of Survivin protein and the expression of Caspase-9 decreased in Hep-2 cells due to the influence of water soluble toad. The result of MTT assay showed that the growth inhibition of Hep-2 cells was enhanced by the aqueous extract of Toadstool with the increase of drug concentration and time.