Netrin-1 induces the anti-apoptotic and pro-survival effects of B-ALL cells through the Unc5b-MAPK axis

作者全名："Huang, Lan; An, Xizhou; Zhu, Yao; Zhang, Kainan; Xiao, Li; Yao, Xinyuan; Zeng, Xing; Liang, Shaoyan; Yu, Jie"

作者地址："[Huang, Lan; An, Xizhou; Zhu, Yao; Zhang, Kainan; Xiao, Li; Yao, Xinyuan; Zeng, Xing; Yu, Jie] Chongqing Med Univ, Dept Hematol & Oncol, Childrens Hosp, 136 Zhongshanerlu, Chongqing 400014, Peoples R China; [Huang, Lan] Minist Educ, Natl Clin Res Ctr Child Hlth & Disorders, Chongqing Key Lab Pediat, Key Lab Child Dev & Disorders, Chongqing, Peoples R China; [Huang, Lan; Zhu, Yao; Zhang, Kainan; Liang, Shaoyan] Chongqing Med Univ, Pediat Res Inst, Childrens Hosp, Chongqing, Peoples R China"

通信作者："Yu, J (通讯作者)，Chongqing Med Univ, Dept Hematol & Oncol, Childrens Hosp, 136 Zhongshanerlu, Chongqing 400014, Peoples R China."

来源：CELL COMMUNICATION AND SIGNALING

ESI学科分类：MOLECULAR BIOLOGY & GENETICS

WOS号：WOS:000841282600001

JCR分区：Q2

影响因子：8.4

年份：2022

卷号：20

期号：1

开始页：　

结束页：　

文献类型：Article

关键词：Netrin-1; B-cell acute lymphoblastic leukemia; Unc5b; FAK; MAPK; Apoptosis

摘要："Background: B-cell acute lymphoblastic leukemia (B-ALL) comprises over 85% of all acute lymphoblastic leukemia (ALL) cases and is the most common childhood malignancy. Although the 5 year overall survival of patients with B-ALL exceeds 90%, patients with relapsed or refractory B-ALL may suffer from poor prognosis and adverse events. The axon guidance factor netrin-1 has been reported to be involved in the tumorigenesis of many types of cancers. However, the impact of netrin-1 on B-ALL remains unknown. Methods: The expression level of netrin-1 in peripheral blood samples of children with B-ALL and children without neoplasia was measured by enzyme-linked immunosorbent assay (ELISA) kits. Then, CCK-8 cell proliferation assays and flow cytometric analysis were performed to detect the viability and apoptosis of B-ALL cells (Reh and Sup B15) treated with exogenous recombinant netrin-1 at concentrations of 0, 25, 50, and 100 ng/ml. Furthermore, co-immunoprecipitation(co-IP) was performed to detect the receptor of netrin-1. UNC5B expression interference was induced in B-ALL cells with recombinant lentivirus, and then CCK-8 assays, flow cytometry assays and western blotting assays were performed to verify that netrin-1 might act on B-ALL cells via the receptor Unc5b. Finally, western blotting and kinase inhibitor treatment were applied to detect the downstream signaling pathway. Results: Netrin-1 expression was increased in B-ALL, and netrin-1 expression was upregulated in patients with high-and intermediate-risk stratification group of patients. Then, we found that netrin-1 induced an anti-apoptotic effect in B-ALL cells, implying that netrin-1 plays an oncogenic role in B-ALL. co-IP results showed that netrin-1 interacted with the receptor Unc5b in B-ALL cells. Interference with UNC5B was performed in B-ALL cells and abolished the antiapoptotic effects of netrin-1. Further western blotting was applied to detect the phosphorylation levels of key molecules in common signaling transduction pathways in B-ALL cells treated with recombinant netrin-1, and the FAK-MAPK signaling pathway was found to be activated. The anti-apoptotic effect of netrin-1 and FAK-MAPK phosphorylation was abrogated by UNC5B interference. FAK inhibitor treatment and ERK inhibitor treatment were applied and verified that the FAK-MAPK pathway may be downstream of Unc5b. Conclusion: Taken together, our findings suggested that netrin-1 induced the anti-apoptotic effect of B-ALL cells through activation of the FAK-MAPK signaling pathway by binding to the receptor Unc5b."

基金机构：Natural Science Foundation of China [81700158]; Chongqing Health Commission and Science and Technology Bureau [2021MSXM231]; Natural Science Fund of Chongqing Science and Technology [cstc2020jcyjmsxmX0295]; General Project of National Clinical Research Center for Child Health and Disorders [NCRCCHD-2020-GP-09]; Medical Research Project of Chongqing Health Commission [2022WSJK005]; Key Projects of Postgraduate AI Medicine R&D Project of Chongqing Medical University [YJSZHYX202103]

基金资助正文：This work was supported by the [Youth Project of Natural Science Foundation of China#1] under Grant [number 81700158]; [Joint Medical Research Project of Chongqing Health Commission and Science and Technology Bureau#2] under Grant [number 2021MSXM231]; [Natural Science Fund of Chongqing Science and Technology Bureau#3] under grant [number cstc2020jcyjmsxmX0295]; [General Project of National Clinical Research Center for Child Health and Disorders#4] under grant [number NCRCCHD-2020-GP-09]; [Medical Research Project of Chongqing Health Commission#5] under grant [number 2022WSJK005] and [Key Projects of Postgraduate AI Medicine R&D Project of Chongqing Medical University#6] under grant [number YJSZHYX202103].