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蛇葡萄素诱导HeLa细胞凋亡的分子机制。

Molecular mechanisms of ampelopsin from induces apoptosis in HeLa cells.

作者信息

Cheng Peipei, Gui Chun, Huang Jing, Xia Ye, Fang Yu, Da Guozheng, Zhang Xiuqiao

机构信息

Department of Pharmacy, Hubei University of Chinese Medicine, Wuhan, Hubei 430065, P.R. China.

出版信息

Oncol Lett. 2017 Sep;14(3):2691-2698. doi: 10.3892/ol.2017.6520. Epub 2017 Jul 4.

Abstract

Ampelopsin (AMP) is an active ingredient of flavonoid compounds that is extracted from Diels et Gilg. The present study aimed at investigating the antitumor activities of AMP and the possible underlying molecular mechanisms in HeLa cells. A total of three types of tumor cell were selected to screen antitumor activities for AMP using the MTT assay. Flow cytometry was used to analyze the cell apoptotic proportion and the cell cycle. Rhodamine 123 staining was used to determine changes in mitochondrial transmembrane potential. Western blot analysis was used to determine the expression of apoptosis-associated proteins. The results of the present study demonstrated that AMP may inhibit the viability of HeLa cells in a dose- and time-dependent manner. Changes in morphology were observed using fluorescence microscopy. In addition, Annexin V-fluorescein isothiocyanate/propidium iodide (PI) double staining revealed that AMP induced apoptosis in a concentration-dependent manner and PI staining indicated that HeLa cells were arrested in S phase. Furthermore, western blot analysis demonstrated that AMP treatment induced apoptosis through activation of caspases 9 and 3, which was validated by the increasing ratio of B-cell lymphoma 2 (Bcl-2)-associated X protein to Bcl-2. Additionally, the loss of mitochondrial transmembrane potential and the release of cytochrome suggested that AMP-induced apoptosis was associated with the mitochondrial pathway. Taken together, these results indicate that AMP may induce apoptosis via the mitochondrial signaling pathway in HeLa cells.

摘要

白藜芦醇(AMP)是从毛叶蛇葡萄中提取的黄酮类化合物的一种活性成分。本研究旨在探讨AMP对人宫颈癌HeLa细胞的抗肿瘤活性及其潜在的分子机制。采用MTT法,共选择三种肿瘤细胞来筛选AMP的抗肿瘤活性。运用流式细胞术分析细胞凋亡比例和细胞周期。使用罗丹明123染色来测定线粒体跨膜电位的变化。采用蛋白质免疫印迹法分析凋亡相关蛋白的表达。本研究结果表明,AMP可能以剂量和时间依赖性方式抑制HeLa细胞的活力。通过荧光显微镜观察到细胞形态发生变化。此外,膜联蛋白V-异硫氰酸荧光素/碘化丙啶(PI)双染显示,AMP以浓度依赖性方式诱导细胞凋亡,PI染色表明HeLa细胞停滞于S期。此外,蛋白质免疫印迹分析表明,AMP处理通过激活半胱天冬酶9和3诱导细胞凋亡,这通过B细胞淋巴瘤-2(Bcl-2)相关X蛋白与Bcl-2的比例增加得到证实。另外,线粒体跨膜电位的丧失和细胞色素的释放表明,AMP诱导的细胞凋亡与线粒体途径有关。综上所述,这些结果表明,AMP可能通过线粒体信号通路诱导HeLa细胞凋亡。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/545a/5588129/ae40fa75946c/ol-14-03-2691-g00.jpg

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