Mechanisms of β-sitosterol on Inhibiting the Proliferation of Hepatocellular Carcinoma Cells by Targeting CDC25B
10.13471/j.cnki.j.sun.yat-sen.univ（med.sci）.20220509.001
- VernacularTitle:β-谷甾醇靶向CDC25B抑制肝癌细胞增殖
- Author:
Kai WANG
1
;
Wei LI
1
;
Zhi-fang LI
1
;
Hui-qiong JI
2
;
Zhi-wei GUO
3
;
Jun-qiu CAO
4
Author Information
1. Department of General Foundation Requisite Teaching， Dazhou Vocational College of Traditional Chinese Medicine， Dazhou 635000， China
2. Affiliated Hospital of Dazhou Center for Disease Control and Prevention， Dazhou 635000， China
3. Department of Medical Imaging， Nanchong Central Hospital， Nanchong 637000， China
4. Department of Hepatobiliary Surgery， West China Hospital， Sichuan University， Chengdu 610000， China
- Publication Type:Journal Article
- Keywords:
hepatocellular carcinoma;
β-sitosterol;
cell division cyclin 25B;
cell proliferation
- From:
Journal of Sun Yat-sen University(Medical Sciences)
2022;43(4):675-684
- CountryChina
- Language:Chinese
-
Abstract:
ObjectiveTo explore the effects and possible actions mechanisms of β-sitosterol on the proliferation of hepatocellular carcinoma （HCC） cells. MethodsThe target of β-sitosterol was predicted by SwissTarget and TargetNet data platforms. The up-regulated gene in GSE101728 HCC data set was analyzed by GEO2R， and the common gene between the target and the up-regulated gene was analyzed. According to the analysis of TCGA_LIHC data matrix， the relationship between cell division cyclin 25B （CDC25B） and clinical staging， prognosis of HCC was obtained. The activities， proliferation， cycles distribution and apoptosis of HCC cells （HepG2， Hep3B） treated with β-sitosterol were detected by CCK-8， colony-forming assay and flow cytometry. The models of nude mice with subcutaneous xenografted tumors in HepG2 were constructed and divided into control group and β-sitosterol group. The β-sitosterol group was intraperitoneally injected with β-sitosterol （50 mg/kg·d）. The volume and mass of tumors were measured. The expression level of Ki67 was detected by immunohistochemistry. The levels of CDC25B protein in HepG2 and Hep3B cell lines treated with different concentrations of β-sitosterol for different action time were detected by Western blot. HepG2 and Hep3B cell lines with stable overexpression of CDC25B were constructed， and then which were treated with β-sitosterol. The activities， proliferation， cycles distribution and apoptosis of HepG2 and Hep3B cell lines were detected by the above-mentioned methods. ResultsThe expression level of CDC25B in HCC tissues was higher than that in para-carcinoma tissues （P<0.05）. The expression level of CDC25B in patients with TNM staging at stage T2-4， clinical staging at stage Ⅱ-Ⅳ， G3-4 differentiation， recurrence and poor prognosis was higher than that with TNM staging at stage T1， clinical staging at stage I， G1-2 differentiation， non-recurrence and good prognosis， respectively （P<0.05）. The overall survival rate in high-expression CDC25B group was lower than that in low-expression group （P<0.05）. The activities of Hep3B and HepG2 cells were decreased with the increase of β-sitosterol dose and the prolongation of action time. The relative number of clone cells and the percentages of cells during S phase in Hep3B （15 μmol/L β-sitosterol） and HepG2 （18 μmol/L β-sitosterol） were significantly lower than those in control group （P<0.05）， while percentage of cells during G0/G1 phase and apoptosis rate were significantly higher than those in control group （P<0.05）. The volume and mass of xenografted tumors， and relative expression level of Ki67 in β-sitosterol group were significantly lower than those in the control group （P<0.05）. The expression of CDC25B protein in Hep3B and HepG2 cells was decreased with the increase of β-sitosterol concentration and the prolongation of action time. The cells growth， relative number of clone cells， and percentages of cells during S phase in Hep3B and HepG2 cells were all significantly higher than those in β-sitosterol group （P<0.05）， while percentage of cells during G0/G1 phase and apoptosis rate were significantly lower than those in β-sitosterol group （P<0.05）. ConclusionThe β-sitosterol can promote apoptosis of HCC cells and inhibited their proliferation. The inhibition of cells proliferation may be related to inhibition of CDC25B protein.
