Toxoplasma gondii RH strain ROP16Ⅰ protein affects proliferation and the cell cycle in MH-S cells through the JAK-STAT3 pathway
10.3969/j.issn.1002-2694.2025.00.019
- VernacularTitle:弓形虫RH株ROP16Ⅰ蛋白通过JAK-STAT3途径对MH-S细胞增殖及细胞周期影响的研究
- Author:
Jia-ming LI
1
;
Tian-tian DANG
1
;
He YIN
1
;
Zhi-jun ZHAO
1
Author Information
1. 宁夏医科大学总医院医学实验中心,银川 750004;宁夏病原微生物重点实验室,银川 750004;宁夏医学检验临床研究中心,银川 750004
- Publication Type:Journal Article
- Keywords:
ROP16 protein;
MH-S cells;
cell cycle;
STAT3;
p-STAT3(Y705)
- From:
Chinese Journal of Zoonoses
2025;41(2):113-120
- CountryChina
- Language:Chinese
-
Abstract:
This study was aimed at investigating the effects and mechanisms of Toxoplasma gondii type Ⅰ(RH strain)ROP16 protein on proliferation and the cell cycle in mouse alveolar macrophage MH-S cells.We constructed a Toxoplasma gondii type Ⅰ(RH)ROP16 overexpression lentivirus,transduced MH-S cells,and then screened cells with puromycin to obtain a cell line stably overexpressing ROP16Ⅰ.RT-qPCR and western blotting were used to verify expression effects,CCK-8 assays were used to detect cell proliferation activity,and flow cytometry was used to detect cell cycle changes.Western blotting and RT-qPCR were used to detect the expression levels of p53,p21,CDK6,Cyclin D1,STAT3,p-STAT3(Y705),and JAK1 proteins or genes,and immunofluorescence was used to detect the expression levels of ROP16Ⅰ and p-STAT3(Y705)and their subcellular co-localization in MH-S cells.ROP16Ⅰ protein and gene expression were detected in MH-S cells transduced with lentivirus for ROP16Ⅰ overexpression.CCK-8 assays revealed that ROP16Ⅰ promoted the proliferation of MH-S cells(P＜0.01)and enhanced cell viability.Flow cytometry revealed that ROP16Ⅰ overexpression decreased the G0/G1 phase and elevated the G2 and S phases of the cell cycle in MH-S cells(P＜0.01 or P＜0.05).Compared with the MH-S cell group and MH-S-empty vector group,the MH-S-ROP16 cell group showed lower expression of p53 and p21 proteins;higher expression of CDK6,Cyclin D1,p-STAT3(Y705),and JAK1 proteins;lower expression of p53 and p21 mRNAs;and higher expression of CDK6 and Cyclin D1 mRNAs(all P＜0.01).Immunofluorescence revealed that ROP16Ⅰ co-localized with p-STAT3(Y705)in the nucleus and surrounding cytoplasm.Therefore,Toxoplasma gondii type Ⅰ(RH)ROP16Ⅰ protein activates the JAK-STAT3 pathway;shortens the G0/G1 phase and lengthens the G2/S phase of the cell cycle;and promotes cell proliferation.These findings provide a theoretical basis for revealing the mechanism of immune evasion of Toxoplasma gondii,and lay a foundation for research on the prevention and treatment of Toxoplasma gondii pneumonia.
