The Mechanism of Methionine Sulfoxide Reductase A in Invasion and Metastasis of Renal Clear Cell Carcinoma
10.13471/j.cnki.j.sun.yat-sen.univ(med.sci).2025.0410
- VernacularTitle:甲硫氨酸亚砜还原酶A在肾透明细胞癌侵袭转移中的作用机制
- Author:
Hongxiang LIU
1
;
Xihai LIU
;
Minjian CHEN
;
Weide ZHONG
Author Information
1. 肇庆市第一人民医院泌尿外科,广东 肇庆 526000
- Publication Type:Journal Article
- Keywords:
clear cell renal carcinoma;
methionine sulfoxide reductase A;
reactive oxygen species;
epithelial-mesenchymal transition;
tumor metastasis
- From:
Journal of Sun Yat-sen University(Medical Sciences)
2025;46(4):639-650
- CountryChina
- Language:Chinese
-
Abstract:
[Objective]This study aims to investigate the effects of methionine sulfoxide reductase A(MSRA)on the proliferation,apoptosis,invasion,and metastasis of clear cell renal cell carcinoma(ccRCC)through cellular and animal experiments to elucidate its potential biological mechanisms,providing new molecular targets and strategies for the treatment of ccRCC.[Methods]MSRA-overexpressing cell lines were constructed with transfection.The effects of MSRA on the proliferation,apoptosis,invasion,and migration of ccRCC cells were assessed through proliferation assays,colony formation assays,apoptosis assays,wound healing assays,and invasion assays.Further,the mechanisms and related signaling pathways were explored via ROS detection,RT-qPCR,and Western blot.Subsequently,a subcutaneous xenograft mouse model was employed to verify the effect of MSRA on ccRCC tumor growth and metastasis at the animal level to explore its potential molecular mechanisms.[Results]The mRNA and protein expressions of MSRA in OS-RC-2 and 786-O cell lines were low(RT-qPCR:0.57±0.09,0.56±0.04;WB:0.26±0.13,0.24±0.09).RT-qPCR and Western blot experiments confirmed the successful construction of OS-RC-2-pLVSO2-MSRA(RT-qPCR:108.04±1.80;WB:117.01±20.19)and 786-O-pLVSO2-MSRA(973.45±51.37;WB:190.34±30.13)overexpressed cell lines,with statistically significant differences(P＜0.001).Proliferation assays showed reduced proliferation in OS-RC-2-pLVSO2-MSRA(72 h:1.246±0.003)and 786-O-pLVSO2-MSRA(72 h:1.468±0.001),with significant differences(P＜0.001).Colony formation assays revealed a decrease in colony numbers in OS-RC-2-pLVSO2-MSRA(0.090±0.002)and 786-O-pLVSO2-MSRA(0.080±0.002),with significant differences(P＜0.001).Apoptosis assays demonstrated increased apoptosis rates in OS-RC-2-pLVSO2-MSRA(2.013±0.116)and 786-O-pLVSO2-MSRA(4.767±0.199),with significant differences(P＜0.001).Wound healing assays indicated less migration distance in OS-RC-2-pLVSO2-MSRA(0.643±0.028)and 786-O-pLVSO2-MSRA(0.603±0.034),with significant differences(P＜0.001).Transwell assays showed a reduction in the numbers of penetrative cells in OS-RC-2-pLVSO2-MSRA(16.80±2.28)and 786-O-pLVSO2-MSRA(21.40±4.78),with significant differences(P＜0.001).Fluorescence assays indicated a decreased ROS levels in OS-RC-2-pLVSO2-MSRA(50.59±6.24)and 786-O-pLVSO2-MSRA(62.87±5.35),with significant differences(P＜0.001).Western blot analysis showed a decrease in the expression of N-cadherin and Vimentin,and an increase in the expression of E-cadherin in OS-RC-2-pLVSO2-MSRA and 786-O-pLVSO2-MSRA(P＜0.001).Western blot analysis revealed a significant decrease in the expression levels of p-ERK1/2 and p-SMAD3 in OS-RC-2-pLVSO2-MSRA and 786-O-pLVSO2-MSRA(P＜0.001).Animal experiments showed reduced tumor volumes in OS-RC-2-pLVSO2-MSRA(155.00±50.46),with significant differences(P＜0.001).Western blot analysis of tumor tissues from animals confirmed the decreased expression of N-cadherin and Vimentin,and the increased expression of E-cadherin,with significant differences(P＜0.001).IHC experiments of OS-RC-2-pLVSO2-MSRA tumors revealed a decrease in the expression of Ki-67,N-cadherin,and Vimentin,and an increase in the expression of E-cadherin,with significant differences(P＜0.001).[Conclusion]MSRA overexpression inhibits ROS expression in ccRCC,suppresses the EMT process,and consequently inhibits the proliferation,invasion,and metastasis of ccRCC.
