| 邱槿怡,黄家明,周毅洁,豆荣昆,毕振飞,杨亚蓝,董银松,向飞丹州,茆灿泉.一种天然植物抗菌液对HEPG-2细胞的杀伤及作用机制研究[J].,2017,17(26):5011-5016 |
| 一种天然植物抗菌液对HEPG-2细胞的杀伤及作用机制研究 |
| The Killing Effects and Preliminary Mechanism of Natural Plant-derived Antimicrobial Solution（PAMs）to Human Liver Cancer HEPG-2 Cells |
| 投稿时间：2016-03-25 修订日期：2016-04-16 |
| DOI：10.13241/j.cnki.pmb.2017.26.003 |
| 中文关键词: 天然植物抗菌液（PAMs） 民族医药 HEPG-2细胞 凋亡途径 |
| 英文关键词: Natural plant-derived antimicrobial solution(PAMs) Folk medicine HEPG-2 cancer cells Apoptosis pathway |
| 基金项目:科技部"自然保护区生物标本标准化整理、整合和共享子项目"资助（2005DKA21404）；云南普洱丹州制药股份有限公司资助项目（VR01HX11131300411130） |
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| 中文摘要: |
| 摘要 目的：采用多种方法探究天然植物抗菌液（PAMs）对人肝癌细胞HEPG-2的杀伤及初步分子作用机理。方法：运用形态学观察、MTT法测定PAMs对HEPG-2的抑制作用，AO/EB细胞双染、Annexin V-FITC/PI流式细胞术检测PAMs作用于HEPG-2细胞后细胞的凋亡情况，再通过qRT-PCR与western blot检测HEPG-2中相关基因在mRNA水平和蛋白水平的表达量变化。最后利用划痕实验和transwell小室检测PAMs对HEPG-2细胞迁移能力的抑制。结果：PAMs可以抑制和杀伤HEPG-2细胞，作用效果呈浓度和时间依赖性，且与细胞的凋亡途径密切相关。PAMs可下调HEPG-2细胞中foxm1、survivin、bcl-2的mRNA表达水平，同时在蛋白水平下调foxm1的表达量，上调p53的表达量。除此之外，PAMs还能够有效地抑制HEPG-2细胞的体外迁移。结论：PAMs杀伤HEPG-2细胞的途径之一是通过下调foxm1、上调p53基因的表达量，从而抑制或减少其下游靶基因survivin和bcl-2的转录表达，促进细胞凋亡。本研究为基于PAMs抗癌创新民族医药的研发提供了重要的前期工作基础。 |
| 英文摘要: |
| ABSTRACT Objective: In this study, a series of experiments were conducted to research the mechanism of anticancer and preliminary molecular effects of PAMs on the HEPG-2 cancer cells. Methods: Morphological observation and MTT assay were used to explore the inhibition and killing effect of PAMs acting on HEPG-2. AO/EB staining and Annexin V-FITC/PI staining were employed to observe the apoptosis of HEPG-2 treated with PAMs. The expression level of Foxm1, bcl-2 and others genes in HEPG-2 cells were detected by using qRT-PCR and western blot. Wound healing and transwell experiments determined if PAMs can inhibit the migration of HEPG-2. Results: PAMs can inhibit and kill HEPG-2 cells in time and dose-dependent manners, and the cytotoxic effects were closely related to the cell apoptosis. The mRNA expression of foxm1,bcl-2 and surviving gene were remarkably decreased in HEPG-2 cells after the treatment of PAMs. PAMs decreased the FoXM1 protein expression in HEPG-2 cells, while up-regulating the p53 protein expression., and it could also inhibit the migration of cancer cells. Conclusion: The possible molecular mechanism for the killing of HEPG-2 cancer cells by PAMs was proposed. By down-regulating the expression of foxm1 and up-regulating the expression of p53, the transcriptional expression of their downstream target genes survivin and bcl-2 was inhibited or reduced, hence enhancing the cancer cell apoptosis. This study provides an important foundation for the development of anti-cancer Chinese folk medicine based on PAMs. |
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