Mechanism of Cantharidin-induced Morphological Changes and Dissociation of HCT116 Cells
10.13422/j.cnki.syfjx.20191024
- VernacularTitle: 斑蝥素诱导HCT116细胞形态改变及脱黏附机制
- Author:
Xin ZHANG
1
;
Hui ZHAO
1
;
Jie WEI
1
;
Jun GUO
1
;
Gang HU
1
;
Ya-jun LIN
1
Author Information
1. Key Laboratory of Geriatrics Under Ministry of Health, National Center of Gerontology, Beijing Hospital, Beijing 100730, China
- Publication Type:Research Article
- Keywords:
cantharidin;
HCT116 cell;
cytoskeleton;
cell adhesion;
RhoA
- From:
Chinese Journal of Experimental Traditional Medical Formulae
2019;25(10):20-25
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To explore the molecular mechanism of cantharidin(CTD) in inducing morphological changes and dissociation in human colon cancer HCT116 cells, in order to study the correlation with tumor metastasis and provide a new basis for clinical use of cantharidin. Method:Different concentrations of CTD (0, 2.5, 5, 10, 15, 20 μmol·L-1) were added to each hole to culture for 7 to 10 days, so as to determine the inhibitory effect of CTD on HCT116 cells; and changes of F-actin cytoskeleton and integrin in HCT116 cells were detected by immunofluorescence staining. Western blot analysis of protein expressions of RhoA,RhoB,RhoC,Cdc42 and Rac1/2/3 of Rho family were performed before and after cantharidin treatment. overexpression of RhoA was constructed by plasmid transient transfection, and effect of 10 μmol·L-1 cantharidin on the morphology and adhesion of overexpressing cells was also observed. Result:Cantharidin induced cytoskeletal remodeling and decreased integrin content in HCT116 colon cancer cells. RhoA protein was a member of Rho enzyme family with the greatest variation after cantharidin action. The proportion of transfected RhoA cells with green fluorescence was about 60%, the expression of RhoA protein in constructed RhoA overexpression cells was significantly increased, compared with wild-type HCT116 cells (P<0.01). However, cantharidin can act on RhoA overexpressed HCT116 cells, reverse its overexpression(P<0.01), and affect its morphology. Conclusion:Cantharidin can inhibit the expression of RhoA protein, induce the morphological changes of HCT116 cells and weaken the adhesion to the basement, thereby inhibiting cell migration.
