Cytogenetics and Molecular Biology Laboratory, Department of Zoology, University of Kalyani, Kalyani 741235, India.
Environ Toxicol Pharmacol. 2013 Jul;36(1):202-14. doi: 10.1016/j.etap.2013.03.023. Epub 2013 Apr 8.
This study tested chemotherapeutic potential of Hydrastis canadensis (HC) extract in HeLa cells in vitro, with emphasis on its drug-DNA interaction and apoptosis induction ability. Nuclear uptake of HC by DAPI, Ao/Eb staining and internucleosomal DNA damage by comet assay was studied through fluorescence microscopy. Possible changes in MMP and apoptotic signalling events were critically analyzed. Cell cycle progression studied through FACS and fragmented DNA through "TUNEL" assay were critically analyzed. RT-PCR studies were conducted for analyzing Cyt-C and Bax translocation in mitochondrial and cytosolic extracts, and Caspase 3 in whole cell lysate. Role of p53-mediated regulation of NF-κβ and TNF-α was elucidated by Western blot analysis. Data of CD and Tm profile of CT-DNA were analyzed. Overall results indicated anti-cancer potential of HC through its ability to induce apoptosis, and interaction with CT-DNA that changed structural conformation of DNA, proving HC to be a promising candidate for chemoprevention.
本研究旨在测试 HC 提取物在 HeLa 细胞中的体外化疗潜力，重点研究其与药物-DNA 的相互作用和诱导细胞凋亡的能力。通过荧光显微镜研究 DAPI、Ao/Eb 染色和彗星试验检测到的 HC 进入细胞核、核内 DNA 片段化和核小体间 DNA 损伤。还对 MMP 和凋亡信号转导事件的可能变化进行了深入分析。通过流式细胞术研究细胞周期进程，通过“TUNEL”检测分析碎片化 DNA。通过 RT-PCR 分析分析线粒体和细胞质提取物中 Cyt-C 和 Bax 的易位以及全细胞裂解物中的 Caspase 3。通过 Western blot 分析阐明了 p53 介导的 NF-κβ和 TNF-α的调节作用。还分析了 CT-DNA 的 CD 和 Tm 谱数据。总体结果表明，HC 通过诱导细胞凋亡和与 CT-DNA 的相互作用具有抗癌潜力，这种相互作用改变了 DNA 的结构构象，证明 HC 是化学预防的有前途的候选药物。
