Targeting microRNA-125b inhibited the metastasis of Alisertib resistance cells through mediating p53 pathway.
10.3760/cma.j.cn112152-20200511-00438
- VernacularTitle:miRNA－125b经p53通路调控Alisertib耐药细胞代谢的机制研究
- Author:
Fu Li YANG
1
;
Xin CHEN
2
;
Fei ZHENG
1
;
Xiang ye LIU
2
;
Na SUN
2
;
Rong Qing LI
2
;
Zhen JIANG
1
;
Jing HAN
2
;
Jing YANG
2
Author Information
1. Department of Pathogenic Biology and Immunology, Xuzhou Medical University, Jiangsu Province Key Laboratory of Immunity and Metabolism, National Experimental Teaching Demonstration Center of Basic Medicine, Xuzhou Medical University, Xuzhou 221000, China.
2. Department of Pathogenic Biology and Immunology, Xuzhou Medical University, Jiangsu Province Key Laboratory of Immunity and Metabolism, National Experimental Teaching Demonstration Center of Basic Medicine, Xuzhou Medical University, Jiangsu International Joint Laboratory for Immunology and Metabolism, Xuzhou 221000, China.
- Publication Type:Journal Article
- Keywords:
Alisertib resistance;
Colon neoplasms;
EMT;
Glycolysis;
MicroRNA-125b;
p53
- MeSH:
Animals;
Female;
Mice;
Azepines;
Cell Line, Tumor;
Cell Movement/genetics*;
Cell Proliferation;
Gene Expression Regulation, Neoplastic;
MicroRNAs/genetics*;
RNA, Messenger;
Tumor Suppressor Protein p53/genetics*;
Humans;
Drug Resistance, Neoplasm
- From:
Chinese Journal of Oncology
2023;45(6):499-507
- CountryChina
- Language:Chinese
-
Abstract:
Objective: To clarify the mechanisms involvement in Alisertib-resistant colorectal cells and explore a potential target to overcome Alisertib-resistance. Methods: Drug-resistant colon cancer cell line (named as HCT-8-7T cells) was established and transplanted into immunodeficient mice. The metastasis in vivo were observed. Proliferation and migration of HCT-8-7T cells and their parental cells were assessed by colony formation and Transwell assay, respectively. Glycolytic capacity and glutamine metabolism of cells were analyzed by metabolism assays. The protein and mRNA levels of critical factors which are involved in mediating glycolysis and epithelial-mesenchymal transition (EMT) were examined by western blot and reverse transcription-quantitative real-time polymerase chain reaction(RT-qPCR), respectively. Results: In comparison with the mice transplanted with HCT-8 cells, which were survival with limited metastatic tumor cells in organs, aggressive metastases were observed in liver, lung, kidney and ovary of HCT-8-7T transplanted mice (P<0.05). The levels of ATP [(0.10±0.01) mmol/L], glycolysis [(81.77±8.21) mpH/min] and the capacity of glycolysis [(55.50±3.48) mpH/min] in HCT-8-7T cells were higher than those of HCT-8 cells [(0.04±0.01) mmol/L, (27.77±2.55) mpH/min and(14.00±1.19) mpH/min, respectively, P<0.05]. Meanwhile, the levels of p53 protein and mRNA in HCT-8-7T cells were potently decreased as compared to that in HCT-8 cells (P<0.05). However, the level of miRNA-125b (2.21±0.12) in HCT-8-7T cells was significantly elevated as compared to that in HCT-8 cells (1.00±0.00, P<0.001). In HCT-8-7T cells, forced-expression of p53 reduced the colon number (162.00±24.00) and the migration [(18.53±5.67)%] as compared with those in cells transfected with control vector [274.70±40.50 and (100.00±29.06)%, P<0.05, respectively]. Similarly, miR-125b mimic decreased the glycolysis [(25.28±9.51) mpH/min] in HCT-8-7T cells as compared with that [(54.38±12.70)mpH/min, P=0.003] in HCT-8-7T cells transfected with control. Meanwhile, in comparison with control transfected HCT-8-7T cells, miR-125b mimic also significantly led to an increase in the levels of p53 and β-catenin, in parallel with a decrease in the levels of PFK1 and HK1 in HCT-8-7T cells (P<0.05). Conclusions: Silencing of p53 by miR-125b could be one of the mechanisms that contributes to Alisertib resistance. Targeting miR-125b could be a strategy to overcome Alisertib resistance.
