Effect of Netrin-1 on VEGFA Expression in T-ALL Cells and Its Related Mechanism.
10.19746/j.cnki.issn.1009-2137.2022.04.012
- Author:
Yao ZHU
1
;
Hai-Yan LIU
2
;
Yan XIANG
1
;
Hui YANG
2
;
Xin-Yuan YAO
2
;
Xi-Zhou AN
2
;
Kai-Nan ZHANG
1
;
Lan HUANG
1
;
Shao-Yan LIANG
3
;
Jie YU
4
Author Information
1. Department of Hematology & Oncology， Children's Hospital Affiliated to Chongqing Medical University， Key Laboratory of Child Development and Disorders of Ministry of Education， National Center for Clinical Medical Research on Child Health and Diseases (Chongqing), Chongqing Key Laboratory of Pediatrics，Chongqing 400010， China,Institute of Pediatrics， Children's Hospital affiliated to Chongqing Medical University, Chongqing 400010， China.
2. Department of Hematology & Oncology， Children's Hospital Affiliated to Chongqing Medical University， Key Laboratory of Child Development and Disorders of Ministry of Education， National Center for Clinical Medical Research on Child Health and Diseases (Chongqing), Chongqing Key Laboratory of Pediatrics，Chongqing 400010， China.
3. Institute of Pediatrics， Children's Hospital affiliated to Chongqing Medical University, Chongqing 400010， China.
4. Department of Hematology & Oncology， Children's Hospital Affiliated to Chongqing Medical University， Key Laboratory of Child Development and Disorders of Ministry of Education， National Center for Clinical Medical Research on Child Health and Diseases (Chongqing), Chongqing Key Laboratory of Pediatrics，Chongqing 400010， China,E-mail: yujie@hospital.cqmu.edu.cn.
- Publication Type:Journal Article
- Keywords:
Akt/mTOR;
Netrin-1;
T Cell Acute Lymphoblastic Leukemia;
Vascular Endothelial Growth Factor-A
- MeSH:
Child;
Humans;
Integrins;
Netrin-1/metabolism*;
Precursor T-Cell Lymphoblastic Leukemia-Lymphoma;
Proto-Oncogene Proteins c-akt/metabolism*;
T-Lymphocytes;
TOR Serine-Threonine Kinases/metabolism*;
Vascular Endothelial Growth Factor A
- From:
Journal of Experimental Hematology
2022;30(4):1049-1055
- CountryChina
- Language:Chinese
-
Abstract:
UNLABELLED:AbstractObjective: To investigate the effect of the axon guidance factor Netrin-1 on the expression of VEGFA in T cell acute lymphoblastic leukemia(T-ALL) and its related mechanism.
METHODS:ELISA assays were applied to detect the levels of Netrin-1 and VEGFA in the bone marrow (BM) samples from children in the T-ALL and control group. The level of Netrin-1 and VEGFA were compared between control children and patients, and the liner correlation between Netrin-1 and VEGFA was analyzed. The T-ALL cells Jurkat and Molt-4 were culture in vitro, and the cells were treated with different concentration of Netrin-1 (0, 25, 50, 100 ng/ml) for 24 h, quantitative RT-PCR (qRT-PCR) and Western blot were used to detect the VEGFA expression in Jurkat, Molt-4 cells. The expression of Netrin-1 receptors in T-ALL cells was detected by qRT-PCR and the interaction between Netrin-1 and receptor in each cells was detected by co-IP. Furthermore, Western blot was used to detect the phosphorylation level of key prateins of AKT signal transduction pathway including Akt and mTOR in T-ALL cells treated with Netrin-1 (100 ng/ml). The expression of VEGFA and phosphorylation of AKT pathway transducers were detected by Western blot, after T-ALL cells treated with Netrin-1 (100 ng/ml) combined with inhibitors specific to Akt or mTOR.
RESULTS:The expression level of Netrin-1 and VEGFA in T-ALL patients BM samples were both signi-ficantly higher than that of control group. And the expression level of Netrin-1 was positively correlated with that of VEGFA（r2=0974). With the increase of Netrin-1 concentration, the expression level of VEGFA also increased(P<0.05）. Netrin-1 interacted with its receptor, integrin-β4 at the Netrin-1 concentration of 100 ng/ml. Further, the treatment of Netrin-1 could increase the phosphorylation of Akt and mTOR, which were the key transducers of AKT pathway. After treatment of T-ALL cells with Netrin-1 (100 ng/mL) and Akt inhibitor, the expression of VEGFA and phosphorylation of Akt or mTOR decreased. When the cells were treated with Netrin-1(100 ng/ml) and mTOR inbititor, the phosphorylation level of mTOR and the expression of VEGFA decreased, the phosphorylation level of Akt increased.
CONCLUSION:The expression of Netrin-1 and VEGFA in bone marrow of childred with T-ALL were abnormal, and there was a linear relationship between them. Netrin-1 can interact with its receptor, integrin-β4 and activate AKT transduction pathway to elevate the expression of VEGFA in T-ALL cells.
