Compound Berberine Induced Autophagyvia P13K/AKT/mTOR Signaling Pathway in CNE2 NPC Cells

Compound Berberine Induced Autophagyvia P13K/AKT/mTOR Signaling Pathway in CNE2 NPC Cells

Objective: To investigate, through an experimental analysis, the autophagy-inducing effect of Compound Berberine (CBBR) on cells of CNE2 nasopharyngeal carcinoma (NPC) and its potential intervening targets in the P13K/AKT/mTOR signaling pathway.

Methods: CNE2 cells were taken as the target cells in this study at the exponential growth level. The concentration of IC50 for CBBR was first calculated by MTT research. Then, for the following intervention studies, 3 different concentrations of CBBR, i.e. 0.25 mg.mL-1, 0.50 mg.mL-1 and 1.00 mg.mL-1 around the IC50 concentration, were taken, respectively. The method of fluorescein labeling was used to test the inducing effect of CBBR on CNE2 cell autophagy. The Western blot technique was followed to investigate the changes of main messenger molecules in the P13K/AKT/mTOR autophagy-related signaling pathway, both combined in a comparative way with the 3-MA block test and carried out by detecting the expressive levels of Beclin 1, LC3-II and LC3-I as well as the LC3-II:LC3-I ratio.

Results: CBBR IC50 at the level of 0.5mg.mL-1 was calculated. The inductive effect of CBBR on the autophagy of CNE2 cells was shown to have occurred in different modes, not just in a simple concentration-dependent tendency, with an effect marginal at 0.25mg.mL-1 concentration and maximal at 0.50mg.mL-1 concentration, while only marginally higher at 1.00mg.mL-1 concentration than at 0.5mg.mL-1 concentration was noticeable. While its inductive impact was slightly reduced by 3-MA after pretreatment, the effect combined with CBBR was still substantially greater than that of 3-MA simply blocked. Furthermore, improvements in the expressive levels of Beclin1, LC3-II and LC3-I as well as LC3-II:LC3-I all showed a pattern corresponding to the altered autophagic characteristics of CNE2 cells (P<0.05 or P<0.01), provided the additional supporting evidence for the inducing effect of CBBR on CNE2 cell autophagy.

Conclusions: CBBR can inhibit the proliferating activity of CNE2 cells by inducing increased autophagous activity through the P13K/AKT/mTOR signaling pathway intermediate targets, and this effect could not be fully blocked by the 3-MA antagonist. Some factors, such as miR-125b via activation of the CXCL 12/CXCR4 axis, could trigger autophagic progress epigenetically as a mediator to cause epithelial-mesenchymal transition (EMT), promote tumor cell invasion, and induce chemical drug resistance by opposites, and CBBR could be used to inhibit the proliferating behavior of NPC cells by interfering in such a way.

Author (s) Details

Shuhua Chen
The Second Municipal Hospital of Foshan, Guangdong, 528000, China.

Yuqin Liu
Huaihua Municipal TCM Hospital, Hunan, 418000, China.

Suhua Yu
Chinese Medicine University of Hunan, Changsha, 410208, China.

Ting Lin
Huaihua Municipal TCM Hospital, Hunan, 418000, China.

Shifan Huang
Huaihua Municipal TCM Hospital, Hunan, 418000, China.

Zhenfeng Zhou
Yueyang Municipal TCM Hospital, Hunan, 414000, China.

Xiang Liu
The Second Municipal Hospital of Foshan, Guangdong, 528000, China.

Yingchun He
Chinese Medicine University of Hunan, Changsha, 410208, China.

Jingying Fan
Chinese Medicine University of Hunan, Changsha, 410208, China.

Daofa Tian
Chinese Medicine University of Hunan, Changsha, 410208, China and The First Affiliated Hospital, Chinese Medicine University of Hunan, Changsha, 410007, China.

View Book :- https://stm.bookpi.org/CTMMR-V12/issue/view/10

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