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中国生物工程杂志

China Biotechnology
China Biotechnology
China Biotechnology  2018, Vol. 38 Issue (4): 17-23    DOI: 10.13523/j.cb.20180403
    
Molecular Mechanism of Inducing 2774-C10 Cell Apoptosis and G1/S Cell Cycle Arrest by Ethanol Extract from Elephantopus mollis H.B.K.
Xiang HUANG1,Jie YANG1,Pei-yan HE1,Zhi-hui WU1,Hui-lan ZENG2,Xin-Ning WANG3(),Jian-wei JIANG1()
1 Medical College,Jinan University,Guangzhou 510632,China
2 The First Affiliated Hospital of Jinan University, Guangzhou 510632,China
3 First Affiliated Hospital of Guangdong Pharmaceutical University, Guangzhou 510080, China
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Abstract   Objective:

To investigate the antitumor mechanism of EM-12, an ethanol extracts from Elephantopus mollis H.B.K., in ovarian cancer.

 Methods:

The effects of EM-12 on the cell viability of 2774-C10 and L02 cells were detected by MTT assay, and colony formation assay. Gene ontology analysis plots for EM-12 regulated gene, as determined by RNA-Seq. Cell cycle was evaluated by PI single staining. Cell apoptosis was evaluated by Annexin V FITC/PI staining. Expressions of apoptosis-relative proteins,and cell cycle-relative in 2774-C10 cells were measured by Western blotting.

 Result:

The cell viability of 2774-C10 was inhibited by EM-12 in a dose manner. Colony formation assays showed that EM-12 decreased colony formation compared with control. Flow cytometry analysis showed an increase of the percentage of apoptotic cells and G1/S phase in a dose-dependent manner treated with EM-12. Gene ontology analysis result showed that the gene set related to G1/S cell cycle arrest scores was enriched in EM-12 treated 2774-C10 cell. EM-12 downregulated the expression of cyclin E2, cyclinD1, CDK2, and CDK6 with different concentrations. In addition, active bands of caspase-3 and PARP could be detected using Western blotting.

 Conclusions:

EM-12 induces the G1/S cell cycle arrest. In addition, EM-12 induces apoptosis by death receptor pathway.

Key words： Elephantopus mollis H.B.K.      Ovarian cancer      Apoptosis      Death receptor pathway      G1/S cell cycle arrest     
Received: 30 October 2017      Published: 08 May 2018
ZTFLH:  Q291  
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Xiang HUANG
Jie YANG
Pei-yan HE
Zhi-hui WU
Hui-lan ZENG
Xin-Ning WANG
Jian-wei JIANG
Cite this article:

Xiang HUANG,Jie YANG,Pei-yan HE,Zhi-hui WU,Hui-lan ZENG,Xin-Ning WANG,Jian-wei JIANG. Molecular Mechanism of Inducing 2774-C10 Cell Apoptosis and G1/S Cell Cycle Arrest by Ethanol Extract from Elephantopus mollis H.B.K.. China Biotechnology, 2018, 38(4): 17-23.

URL:

https://manu60.magtech.com.cn/biotech/10.13523/j.cb.20180403     OR     https://manu60.magtech.com.cn/biotech/Y2018/V38/I4/17

Fig.1 The chemical structure of EM-12 monomer
Fig.2 Effect of EM-12 on cell viability of 2774-C10 cell
Fig.3 Effect of EM-12 on colon formation of 2774-C10 cell
(a) EM-12 reduced colon formation of 2774-C10 cells (b)EM-12 reduced cell number of monoclones (c) Statistical analysis of colon formation (d) Statistical analysis of cell number of monoclones
Fig.4 Effects of EM-12 on G1/S cell cycle arrest of 2774-C10 cell
(a)Gene ontology analysis result showed that gene set related to G1/S cell cycle arrest scores was enriched in EM-12 treated 2774-C10 cell (b) Flow cytometry analysis showed an increase of the percentage of G1/S phase in a dose-dependent manner treated with EM-12
Fig.5 Effects of EM-12 on apoptosis of 2774-C10 cell
Fig.6 The expression of G1/S cell cycle arrest-related proteins detected by the Western blotting
(a) G1/S cell cycle arrest-related proteins were detected by the Western blotting (b)~(f) Grey statistics of Western blotting
Fig.7 The expression of apotosis-related proteins detected by the Western blotting
(a) Apotosis-related proteins were detected by the Western blotting (b)~(e) Grey statistics of Western blotting
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