Effect of Total Flavone of Litchi Semen on Proliferation， Migration， and Invasion of HepG2 Cells Based on JAK2/STAT3 Signaling Pathway
10.13422/j.cnki.syfjx.20221522
- VernacularTitle:基于JAK2/STAT3信号通路探讨荔枝核总黄酮对HepG2增殖、迁移与侵袭的影响
- Author:
Minhang LI
1
;
Xiaocong MA
1
;
Yan TANG
1
;
Jingyun LIANG
1
;
Weisheng LUO
1
;
Xuping HUANG
2
Author Information
1. Guangxi University of Chinese Medicine， Nanning 530001， China
2. Ruikang Hospital Affiliated to Guangxi University of Chinese Medicine， Nanning 530001， China
- Publication Type:Journal Article
- Keywords:
total flavone of Litchi Semen;
Janus kinase 2/signal transducer and activator of transcription 3 （JAK2/STAT3） signaling pathway;
invasion;
migration;
HepG2;
epithelial-mesenchymal transition（EMT）
- From:
Chinese Journal of Experimental Traditional Medical Formulae
2022;28(22):85-92
- CountryChina
- Language:Chinese
-
Abstract:
ObjectiveTo study the effect of total flavone of Litchi Semen （TFL） on proliferation， apoptosis， migration， and invasion of hepatoma cells HepG2. MethodMethyl thiazolyl tetrazolium colorimetric （MTT） assay was used to detect the effect of different-dose TFL and cisplatin on the proliferation of HepG2 cells. TdT-mediated dUTP nick-end labeling （TUNEL） assay was used to detect the effects of low， medium， and high-dose （70， 140， 210 mg·L-1） of TFL and cisplatin （60 mg·L-1） on the apoptosis of HepG2 cells， thus selecting the optimal dose of TFL for the follow-up experiment. HepG2 cells were divided into a blank group， a TFL group （140 mg·L-1）， a TFL+XL019 group （140 mg·L-1 TFL+0.5 μmol·L-1 XL019）， and a TFL+TPI-1 group （140 mg·L-1 TFL+1 μmol·L-1 TPI-1）. The effect of TFL on migration and invasion of HepG2 cells were examined by wound healing test and Transwell invasion assay， and the effect of TFL on the expression of epithelial-mesenchymal transition （EMT） marker in HepG2 cells were examined by cell immunofluorescence assay. Western blot was used to detect the expression of key proteins in Janus kinase 2（JAK2）/signal transducer and activator of transcription 3 （STAT3） signaling pathway after the intervention by TFL. ResultMTT assay showed that the proliferation of HepG2 cells was significantly inhibited by TFL and cisplatin at 24 and 48 h as compared with blank group （P<0.01）， and the half maximal inhibitory concentration （IC50） of TFL on HepG2 cells was （136.7±2.40） mg·L-1 at 24 h and （106.8±1.11） mg·L-1 at 48 h. The IC50 of cisplatin on HepG2 cells was （58.48±2.04） mg·L-1 at 24 h and （5.15±0.56） mg·L-1 at 48 h. The results of TUNEL assay showed that TFL induced apoptosis of HepG2 cells. The optimal dose of TFL was 140 mg·L-1. The results of wound healing test showed that compared with the blank group， the TFL group， TFL+XL019 group， and the TFL+TPI-1 group significantly inhibited the migration of HepG2 cells （P<0.05， P<0.01）. As compared with the TFL group， the inhibitory effect of the TFL+XL019 Group was significantly increased （P<0.05）， while that of the TFL+TPI-1 group was significantly decreased （P<0.01）. The Transwell invasion assay showed that compared with the blank group， the TFL group， TFL+XL019 group， and the TFL+TPI-1 group significantly inhibited the invasion of HepG2 cells （P<0.01）. As compared with the TFL group， the inhibitory effect of the TFL+XL019 group was significantly increased （P<0.05）， while that of the TFL+TPI-1 group was significantly decreased （P<0.01）. The results of immunofluorescence showed the intervention of TFL up-regulated the expression of E-cadherin， and down-regulated the expression of Vimentin in HepG2 cells， which was stronger in the TFL+XL019 group and weaker in the TFL+TPI-1 group. The results of Western blot showed that compared with the blank group， the TFL group， TFL+XL019 group， and the TFL+TPI-1 group did not affect the expression of JAK2 or STAT3 protein， but significantly decreased the expression levels of phosphorylatied （p）-JAK2 and p-STAT3 （P<0.05， P<0.01）. As compared with the TFL group， the expression levels of p-JAK2 and p-STAT3 in the TFL+XL019 group were significantly decreased （P<0.01）， while those in the TFL+TPI-1 group were significantly increased （P<0.01）. Compared with the blank group， the TFL group significantly increased the expression level of Src-homology domain 2 containing protein tyrosine phosphatase-1（SHP-1） with sh2 domain （P<0.01）. ConclusionTFL has the effects of inhibiting the proliferation， promoting apoptosis of HepG2 cells， and reversing the EMT process of HepG2 cells to reduce the migration and invasion， which are presumably related to the activation of SHP-1 by TFL to block JAK/STAT3 signaling pathway.
