扶正抗癌方通过PTEN/PI3K/Bad通路调控肺癌A549细胞增殖与凋亡

黎金华; 吴万垠; 杨小兵

doi:10.13422/j.cnki.syfjx.2017210098

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扶正抗癌方通过PTEN/PI3K/Bad通路调控肺癌A549细胞增殖与凋亡

药理 | 更新时间：2024-01-04

- 扶正抗癌方通过PTEN/PI3K/Bad通路调控肺癌A549细胞增殖与凋亡
- Effect of FZKA Decoction in Regulating A549 Proliferation and Apoptosis PTEN/PI3K/Bad
- 中国实验方剂学杂志 2017年23卷第21期页码：98-103
- 作者机构：
- 作者简介：
- 基金信息：
  
  国家自然科学基金项目（81503507，81273965）;广东省自然科学基金项目（2015A030310245）;广东省建设中医药强省科研课题项目（20141104）
- DOI：10.13422/j.cnki.syfjx.2017210098
  中图分类号： R285
- 纸质出版日期：2017
- 稿件说明：
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黎金华, 吴万垠, 杨小兵. 扶正抗癌方通过PTEN/PI3K/Bad通路调控肺癌A549细胞增殖与凋亡[J]. 中国实验方剂学杂志, 2017,23(21):98-103.

LI Jin-hua, WU Wan-yin, YANG Xiao-bing. Effect of FZKA Decoction in Regulating A549 Proliferation and Apoptosis PTEN/PI3K/Bad[J]. Chinese journal of experimental traditional medical formulae, 2017, 23(21): 98-103.
黎金华, 吴万垠, 杨小兵. 扶正抗癌方通过PTEN/PI3K/Bad通路调控肺癌A549细胞增殖与凋亡[J]. 中国实验方剂学杂志, 2017,23(21):98-103. DOI： 10.13422/j.cnki.syfjx.2017210098.

LI Jin-hua, WU Wan-yin, YANG Xiao-bing. Effect of FZKA Decoction in Regulating A549 Proliferation and Apoptosis PTEN/PI3K/Bad[J]. Chinese journal of experimental traditional medical formulae, 2017, 23(21): 98-103. DOI： 10.13422/j.cnki.syfjx.2017210098.

摘要

目的：观察扶正抗癌（FZKA）方对肺癌细胞A549增殖的调控作用，探讨其作用的分子机制。方法：以非小细胞肺癌细胞A549为研究对象，以FAKA方0.8，1.2，1.6，2.0，2.4，2.8，3.2 g·L-1干预，空白组，分别孵育24，48，72 h后，设立采用噻唑蓝（MTT）比色法检测FZKA方对A549细胞活力的影响；以FAKA方1.2，1.6，2.0 g·L-1干预，设立空白组，培养24 h后，采用实时荧光定量聚合酶链式反应（Real-time PCR）检测FZKA方对A549细胞人第10号染色体同源丢失性磷酸酶-张力蛋白（phosphatase and tensin homolog deleted on chromosome ten，PTEN）mRNA水平的影响；孵育24 h后，采用蛋白质免疫印迹法（Western blot）分析FZKA方对PTEN，磷脂酰肌醇3-激酶（phosphatidylinositol-3-kinase，PI3K），磷酸化的促凋亡蛋白Bad（Phospho-Bad，p-Bad）表达的影响并探讨其相关性。结果：与空白组比较，FZKA方能明显抑制A549细胞增殖，处理24 h后，FZKA从0.8 g·L-1开始，处理48，72 h后，FZKA方从0.4 g·L-1开始，细胞存活率明显降低（P < 0.05，P < 0.01）；与空白组比较，处理24 h后，FZKA方从0.8 g·L-1开始以浓度依赖性上调PTEN mRNA和蛋白的表达（P < 0.05，P < 0.01），FZKA方从1.6 g·L-1开始以浓度依赖性下调PI3K蛋白表达（P < 0.05，P < 0.01），从2 h开始以时间依赖性上调p-Bad蛋白的表达（P < 0.05，P < 0.01）。结论：FZKA方可通过上调PTEN来调控下游PI3K/Akt/Bad通路相关蛋白的表达进而抑制A549的增殖，促进肺癌细胞凋亡。

Abstract

Objective: To observe the effect of Fuzheng Kangai (FZKA) decoction on the proliferation and apoptosis

in order to discuss the molecular mechanism of apoptosis induced by FZKA decoction in A549 cells. Method: Non-small lung cancer cells (A549 cells) were taken as the research objects and treated for 24

72 h with 0 (blank group)

0.8

1.2

1.6

2.0

2.4

2.8

3.2 g·L-1 FAKA decoction. Then

methyl-thiazolyl-tetrazolium (MTT) assay was used to detect the effect of FZKA decoction on the cell proliferation and apoptosis. A549 cells were treated for 24 h with 0 (blank group)

1.2

1.6

2.0 g·L-1. Then

quantitative Real-time PCR was used to detect the effect of FZKA decoction on PTEN mRNA expression. A549 cells were treated for 24 h with 0 (blank group)

0.8

1.2

1.6

2.0 g·L-1. Then

Western blot assay was used to detect the expressions of phosphatase and tensin homolog deleted on chromosome ten (PTEN)

phosphatidylinositol-3-kinase (PI3K) and phospho-Bad (p-Bad)

and discuss the relationship between them. Result: Compared with blank group

the proliferation of A549 cells were inhibited by FZKA decoction in concentration-dependent and time-dependent manners (P < 0.01). Compared with blank group

mRNA and protein expressions of PTEN were up-regulated in a concentration-dependent manner (P < 0.05

P < 0.01). Compared with blank group

the protein expression of PI3K was down-regulated in a concentration-dependent manner (P < 0.05

P < 0.01)

and p-Bad was down-regulated in a time-dependent manner (P < 0.05

P < 0.01). Conclusion: Apoptosis is induced by FZKA decoction by up-regulating PTEN and regulating PI3K/Akt/Bad pass way in A549 cells.

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