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| Pp-10, a Monomer Purified from Paris polyphylla, Induces Apoptosis and Autophagy of Human Gastric Carcinoma BGC-823 Cells by Inhibiting PI3K/Akt Signaling Pathway |
| WANG Juan-juan1, ZHANG Peng1, HUANG Zhi-hong2, CHEN Jia-jin1, LI Qiang1, WANG Guo-cai3, LI Yao-lan3, JIANG Jian-wei1 |
1. Department of Biochemistry, Medical College, Jinan University, Guangzhou 510630, China;
2. Department of Pulmonary, The First Affiliated Hospital, Jinan University, Guangzhou 510632, China;
3. Institute of Traditional Chinese Medicine and Nature Products, College of Pharmacy, Guangzhou 510630, China |
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Abstract Objective: To investigate pp-10, a monomer purified from Paris Polyphylla, induced apoptosis of human gastric carcinoma BGC-823 cells and the possible mechanism behind this effect. Methods: The effects of pp-10 on the proliferation of BGC-823 Cells were evaluated by the MTT assays and colony formation assays.The cellular morphology changes of BGC-823 cells were observed by fluorescence microscopy after Hoechst 33342 staining. Cell apoptosis was detected by Annexin V-FITC/Propidium iodide (PI) double staining using flow cytometry. The expression of apoptotic and autophagy associated proteins as LC3,P62,Caspase-9, Caspase-3, the Caspase substrate poly(ADP-ribose) polymerase (PARP), Bcl-2 proteins family such as Bax, Bcl-2, PI3k/Akt signaling pathway numbers such as Akt, p-Akt, mTOR, p-mTOR, P70s6k and p-P70s6k were detected by Western blotting. Results: Paris Polyphylla monomer pp-10 inhibited the proliferation of BGC-823 cells significantly in dose- and time- dependent manner (P<0.05). Cell clone formation inhibition assay demonstrated that cell clones decreased with the increase of drug concentrations. Apoptotic morphology such as cell shrinkage, nuclear condensation, nuclear fragmentation, chromatin condensation were also observed by staining with Hoechst33342 under fluorescence microscope. Flow cytometry analysis showed an increase of the percentage of apoptotic cells in a dose-dependent manner treated with pp-10 for 24 h. Active bands of mitochondria associated apoptotic signaling pathway proteins such as Caspase-3, Caspase-9 and PARP could be seen with western blotting analysis after BGC-823 cells were incubated with different concentrations of pp-10 for 24 h. In addition, pp-10 upregulated the expression of Bax, a pro-apoptotic protein, down-regulated the expression of Bcl-2 protein, an anti-apoptotic protein, up-regulated the expression of LC3, down-regulated the expression of P62 protein,and also down-regulated PI3K/Akt signal pathway as decreasing the phosphorylation of p-mTor, p-p70S6K, downstream of Akt. Conclusions: pp-10 inhibits cell proliferation and induces the apoptosis and autophagy of human Gastric carcinoma BGC-823 cells by activation of Bax, down-regulation of Bcl-2 and reduction of PI3K/Akt signaling pathway.
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Received: 18 November 2014
Published: 25 February 2015
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Cite this article:
WANG Juan-juan, ZHANG Peng, HUANG Zhi-hong, CHEN Jia-jin, LI Qiang, WANG Guo-cai, LI Yao-lan, JIANG Jian-wei. Pp-10, a Monomer Purified from Paris polyphylla, Induces Apoptosis and Autophagy of Human Gastric Carcinoma BGC-823 Cells by Inhibiting PI3K/Akt Signaling Pathway. China Biotechnology, 2015, 35(2): 31-37.
URL:
https://manu60.magtech.com.cn/biotech/10.13523/j.cb.20150205 OR https://manu60.magtech.com.cn/biotech/Y2015/V35/I2/31
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