Anti-hepatocarcinoma HepG2 Cell Mechanism of Jaranol： Based on PI3K/Akt Signaling Pathway
10.13422/j.cnki.syfjx.20220799
- VernacularTitle:基于PI3K/Akt信号通路探讨华良姜素抗肝癌细胞HepG2凋亡的作用机制
- Author:
Hong YAO
1
;
Yu-xin HOU
1
;
Jin-hong REN
1
;
Yu-xuan WANG
1
;
Hui-qing XUE
1
;
Qing-shan LI
1
Author Information
1. Shanxi Key Laboratory of Innovative Drugs for the Treatment of Serious Diseases Basing on the Chronic Inflammation， Shanxi University of Chinese Medicine， Taiyuan 030619， China
- Publication Type:Journal Article
- Keywords:
Jaranol;
HepG2;
apoptosis;
transcriptome;
phosphatidylinositol 3-kinase/protein kinase B （PI3K/Akt） signaling pathway
- From:
Chinese Journal of Experimental Traditional Medical Formulae
2022;28(9):80-87
- CountryChina
- Language:Chinese
-
Abstract:
ObjectiveTo study the in vitro anti-hepatocarcinoma HepG2 cell mechanism of Jaranol. MethodThe methyl thiazolyl tetrazolium （MTT） assay was employed to examine the inhibition of Jaranol （0， 5， 10， 25， 50， 100， 150， 300 μmol·L-1） on HepG2 cell proliferation at different time （24 ， 48 ， 72 h）， annexin V-fluorescein isothiocyante/propidium iodide （Annexin V-FITC/PI） kit to detect the effect of Jaranol （0， 3， 15， 75 μmol·L-1） on HepG2 cell apoptosis， and Western blot to determine the influence of Jaranol on the expression of B-cell lymphoma 2 （Bcl-2） and Bcl-2-associated X protein （Bax） in HepG2 cells. Transcriptome sequencing was performed to analyze the differential expression of genes and changes of related signaling pathways after the treatment of HepG2 cells with Jaranol （15 μmol·L-1）. Real-time PCR was carried out to verify the relative mRNA content of differential genes ［TEK， platelet-derived growth factor receptor α （PDGFRA）， spleen tyrosine kinase （SYK）， phosphatidylinositol-4，5-bisphosphate 3-kinase catalytic subunit gamma （PIK3CG）， Janus kinase 3 （JAK3）， membrane-associated guanylate kinase inverted 2 （MAGI2）］. ResultCompared with the blank group， Jaranol decreased HepG2 proliferation （P<0.05， P<0.01）， increased apoptosis rate of HepG2 cells （P<0.05， P<0.01）， raised Bax expression （P<0.05， P<0.01）， and reduced Bcl-2 expression （P<0.05， P<0.01）. Transcriptome sequencing yielded 59 000 regulated genes， 125 of which showed significantly different expression， with 47 up-regulated and 74 down-regulated. Kyoto Encyclopedia of Genes and Genomes （KEGG） analysis showed that the differential genes related to apoptosis in the phosphatidylinositol 3-kinase/protein kinase B （PI3K/Akt） signaling pathway changed significantly after drug addition. The mRNA expression of TEK， PDGFRA， SYK， PIK3CG， JAK3， and MAGI2 decreased in Jaranol （15 μmol·L-1） group compared with that in the control group （P<0.05）. ConclusionIn vitro cytological experiment verified that Jaranol inhibited the proliferation of HepG2 cells and promoted the apoptosis， possibly by influencing the expression of some differential genes in the PI3K/Akt signaling pathway. The result lays an experimental basis for the follow-up study of the anti-tumor effect of Jaranol， and the further development and utilization of flavonoids.
