Effect of Compound Phyllanthus urinaria Ⅱ(CPU Ⅱ) on Proliferation, Apoptosis and Autophagy of Hepatocellular Carcinoma Huh7 Cells
10.13422/j.cnki.syfjx.20182422
- VernacularTitle: 叶下珠复方Ⅱ号对肝癌Huh7细胞增殖、凋亡与自噬的影响
- Author:
Xin-yun DU
1
;
Chang-qing LI
1
;
Bing CHEN
1
;
Xiao-hui LI
1
Author Information
1. Institute of Tropical Medicine, School of Chinese Materia Medica, Guangzhou University of Chinese Medicine, Guangzhou 510405, China
- Publication Type:Research Article
- Keywords:
Phyllanthus urinaria Ⅱ;
autophagy;
proliferation;
apoptosis;
Huh7 cells
- From:
Chinese Journal of Experimental Traditional Medical Formulae
2019;25(2):48-54
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To observe the effect and investigate the mechanism of compound Phyllanthus urinaria Ⅱ(CPU Ⅱ)on proliferation, apoptosis and autophagy of human hepatoma cell line Huh7. Method:Huh7 cells were cultured in vitro and divided into blank control group, high-dose CPU Ⅱ group (40 g·L-1),low-dose CPU Ⅱ group (20 g·L-1), and 5-FU group (0.04 g·L-1). Methye thiazolye telrazlium(MTT) assay was used to detect the inhibitory effect of CPU Ⅱ on proliferation of human hepatoma Huh7 cells. The apoptosis rate was observed by Annexin V-FITC/PI flow cytometry; the changes of autophagosomes in each group were observed by monodansylcadaverin (MDC) staining; Real-time fluorescence quantitative polymerase chain reaction(Real-time PCR)was used to detect the mRNA expression of phosphatidylinositol 3-kinase (PI3K), Serine-threonine protein kinase 2(Akt2), B-cell lympoma-2(Bcl-2) and Microtubule-associated protein 1 light chian 3(LC3Ⅱ) and Western blot was used to detect the protein expression of Akt2 and LC3Ⅱ. Result:CPU Ⅱ(40, 20 g·L-1) significantly inhibited the hepatoma cell line Huh7 and induced apoptosis, with an apoptosis rate of 51.72% and 19.74% respectively, significantly higher than that of control group (P<0.01). After MDC staining, it was observed that a large number of fluorescent particles were distributed in the cytoplasm and nucleus of liver cancer cells under the effect of CPU Ⅱ. As compared with blank control group, the mRNA expression levels of PI3K, Akt2, and Bcl-2 were decreased significantly while the mRNA expression of LC3Ⅱ was increased significantly in CPU Ⅱ high-and low-dose groups (P<0.01); the protein expression of Akt was significantly decreased while protein expression of LC3Ⅱwas significantly increased in CPU Ⅱ high-dose and low-dose groups (P<0.05,P<0.01). Conclusion:CPUⅡ had obvious inhibitory effect on the proliferation of hepatoma cell line Huh7, and the mechanism may be related to inhibiting the activation of PI3K/Akt signaling pathway to induce apoptosis and autophagy.
