Effect of Jiawei Sijunzi Decoction Containing Serum on Human Hepatocellular Carcinoma Hep-G2 Cells

SHI Sheng-ying; LIN Hai-zhen; ZHOU Wei; WANG Su-li; LIN Jing-ming

doi:10.13422/j.cnki.syfjx.2016180088

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Effect of Jiawei Sijunzi Decoction Containing Serum on Human Hepatocellular Carcinoma Hep-G2 Cells

更新时间：2024-01-04

- Effect of Jiawei Sijunzi Decoction Containing Serum on Human Hepatocellular Carcinoma Hep-G2 Cells
- Chinese Journal of Experimental Traditional Medical Formulae Vol. 22, Issue 18, Pages: 88-93(2016)
- 作者机构：
- 作者简介：
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- DOI：10.13422/j.cnki.syfjx.2016180088
  CLC： R285
- Published：2016
- 稿件说明：
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SHI Sheng-ying, LIN Hai-zhen, ZHOU Wei, et al. Effect of Jiawei Sijunzi Decoction Containing Serum on Human Hepatocellular Carcinoma Hep-G2 Cells[J]. Chinese journal of experimental traditional medical formulae, 2016, 22(18): 88-93.
DOI：

SHI Sheng-ying, LIN Hai-zhen, ZHOU Wei, et al. Effect of Jiawei Sijunzi Decoction Containing Serum on Human Hepatocellular Carcinoma Hep-G2 Cells[J]. Chinese journal of experimental traditional medical formulae, 2016, 22(18): 88-93. DOI： 10.13422/j.cnki.syfjx.2016180088.

摘要

目的：探讨加味四君子汤含药血清对肝癌Hep-G2细胞增殖、凋亡、周期的影响及其作用机制。方法：不同浓度加味四君子汤含药血清处理Hep-G2细胞后，采用细胞活性检测试剂盒（CCK-8）检测细胞增殖；AnnexinV/碘化丙啶（PI）流式细胞术检测细胞凋亡率和周期；hoechst33342荧光染色观察细胞凋亡形态；免疫印迹法（Western blot）检测细胞中磷脂酰肌醇3-激酶（PI3K）/蛋白激酶B（Akt）/雷帕霉素靶蛋白（mTOR）信号通路相关蛋白表达水平。结果：加味四君子汤含药血清呈浓度依赖性抑制肝癌Hep-G2细胞的增殖，诱导Hep-G2细胞凋亡，阻滞细胞于G0/G1期。hoechst33342染色后，随着含药血清浓度的增加，Hep-G2细胞核显著呈碎块状致密浓染。同时，加味四君子汤含药血清能够抑制Hep-G2细胞Akt，mTOR，核糖体S6蛋白激酶（S6），真核翻译起始因子4E结合蛋白1（4EBP1）的磷酸化，从而上调Bcl-2相关X蛋白（Bax）和下调细胞周期蛋白（CyclinD1），B淋巴细胞瘤-2（Bcl-2）的表达。加味四君子汤含药血清与300 nmol·L-1的PI3K/mTOR双重抑制剂VS-5584联合使用具有协同作用，含药血清能增强PI3K/mTOR双重抑制剂VS-5584对Hep-G2细胞中PI3K/Akt/mTOR信号通路靶点Akt和mTOR磷酸化的抑制作用。结论：加味四君子汤含药血清能抑制Hep-G2细胞的增殖，诱导其凋亡，阻滞其于G0/G1期，其机制可能通过阻断PI3K/Akt/mTOR信号通路而实现。

Abstract

Objective: To investigate the effects of Jiawei Sijunzi decoction containing serum on proliferation

apoptosis

and cell cycle of human hepatocellular carcinoma Hep-G2 cells and explore its action mechanism. Method: After Hep-G2 cells were processed with different concentrations of Jiawei Sijunzi decoction containing serum

cell counting kit-8(CCK-8) was used to measure the proliferation of Hep-G2 cells

apoptosis rate and cell cycle were detected by AnnexinV/propidium iodide(PI) flow cytometry. Apoptotic morphology was determined by hoechst33342 staining. The expression levels of related proteins of phosphatidylinositol 3-kinase(PI3K)/protein kinase B(Akt)/mammalian target of rapamycin(mTOR) signaling pathway in Hep-G2 cells were evaluated by Western blot assay. Result: Jiawei Sijunzi decoction containing serum inhibited the proliferation and induced apoptosis of human hepatoma Hep-G2 cells in a dose-dependent manner. Cells were arrested in G0/G1 phase. Hoechst33342 staining results showed prominent morphological changes with chromatin condensation

fragmentation and formation of apoptotic bodies in dose-dependent manner. Western blot analysis showed that Jiawei Sijunzi decoction containing serum could inhibit phosphorylation of Akt

mTOR

ribosomal S6 kinase(S6)

and eukaryotic translation initiation factor 4E-binding protein 1(4EBP1)

up-regulate the expression levels of Bax

and down-regulate the expression levels of CyclinD1

Bcl-2. Jiawei Sijunzi decoction containing serum and PI3K/mTOR dual inhibitor VS-5584 had a synergism effect

and Jiawei Sijunzi decoction containing serum enhanced the inhibitory effects on phosphorylation of Akt and mTOR. Conclusion: Jiawei Sijunzi decoction containing serum could inhibit the proliferation of Hep-G2 cells

induce apoptosis and arrest Hep-G2 cells in G1 phase. This inhibitory effect may be induced by blocking the PI3K/Akt/mTOR pathway.

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