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评价 叶水提物对人慢性髓系白血病细胞(K562 细胞)中某些凋亡相关基因和蛋白的影响。

Evaluation of some genes and proteins involved in apoptosis on human chronic myeloid leukemia cells (K562 cells) by leaves aqueous extract.

机构信息

Faculty of Medicine, Department of Biochemistry, Birjand University of Medical Sciences, Birjand, Iran.

Department of Biology, University of Sistan and Baluchestan, Zahedan, Iran.

出版信息

J Biomol Struct Dyn. 2020 Oct;38(16):4838-4849. doi: 10.1080/07391102.2019.1691661. Epub 2019 Nov 27.

Abstract

Datura innoxia () has an extensive usage in traditional medicine and can also be used for intervention therapy in order to treat cancer. Despite of accomplishing some researches on mechanism, still our knowledge is very little about exact apoptotic mechanism on human chronic myeloid leukemia cells (K562 cells). This study purpose was to clarify the molecular mechanism of apoptosis, which was mediated by leaves aqueous extract in K562 cells. MTT assay and flow cytometry was applied in order to assess the viability and apoptosis induction of K562 cells and normal human lymphoid B cells in the presence. Finally, the expression of the apoptotic related genes (p53, BAX, BCL2, Caspases 3, 6, 7 and 9) were evaluated using quantitative Real-Time PCR. Western blot analysis was applied for assessing the protein expression. MTT results indicated that could inhibit the viability of K562 cells in a dose- and time-dependent manner. In parallel, inhibitory effect on normal human lymphoid B cells was lower in comparison with its effect on K562 cells at the same concentrations and same incubation time. Apoptosis induction in K562 cells after exposure was determined by flow cytometry. Apoptosis was activated by in K562 cells throughout increasing the expression of P53, BAX/BCL2 ratio, caspase 9, 3, 6, 7. Western blot analysis demonstrated significant increase in cleaved PARP-1 and cleaved caspase 3 in treated K562 cells with high leaves aqueous extract concentration. leaves trigger apoptosis in K562 cells throughout intrinsic apoptotic pathway.Communicated by Ramaswamy H. Sarma.

摘要

曼陀罗()在传统医学中有广泛的应用,也可用于干预治疗癌症。尽管已经对其机制进行了一些研究,但我们对其在人类慢性髓性白血病细胞(K562 细胞)的确切凋亡机制仍知之甚少。本研究旨在阐明曼陀罗叶水提物在 K562 细胞中诱导细胞凋亡的分子机制。MTT 法和流式细胞术用于评估 K562 细胞和正常人淋巴细胞 B 细胞在存在下的活力和凋亡诱导。最后,采用实时定量 PCR 评估凋亡相关基因(p53、BAX、BCL2、Caspases 3、6、7 和 9)的表达。采用 Western blot 分析评估蛋白表达。MTT 结果表明,曼陀罗叶水提物可剂量依赖性和时间依赖性地抑制 K562 细胞的活力。同时,在相同浓度和相同孵育时间下,曼陀罗叶水提物对正常人类淋巴细胞 B 细胞的抑制作用低于对 K562 细胞的抑制作用。用流式细胞术测定 K562 细胞暴露后的凋亡诱导情况。凋亡在 K562 细胞中被激活,通过增加 P53、BAX/BCL2 比值、caspase 9、3、6、7 的表达。Western blot 分析表明,高浓度曼陀罗叶水提物处理的 K562 细胞中 cleaved PARP-1 和 cleaved caspase 3 的表达明显增加。曼陀罗叶通过内在凋亡途径诱导 K562 细胞凋亡。由 Ramaswamy H. Sarma 传递。

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