IL-2 Combined with IL-15 Enhanced the Expression of NKG2D Receptor on Patient Autologous NK Cells to Inhibit Wilms' Tumor via MAPK Signaling Pathway

作者全名："Li, Yanping; Li, Yonglin; Xiang, Bin; Tian, Xiaomao; Shi, Qinlin; Liu, Feng; Lin, Tao; Wei, Guanghui"

作者地址："[Li, Yanping; Li, Yonglin; Xiang, Bin; Tian, Xiaomao; Shi, Qinlin; Liu, Feng; Lin, Tao; Wei, Guanghui] Chongqing Med Univ, Childrens Hosp, Key Lab Child Dev & Disorders, Minist Educ,Chongqing Key Lab Pediat,Pediat Res In, Chongqing 400014, Peoples R China; [Shi, Qinlin; Liu, Feng; Lin, Tao; Wei, Guanghui] Chongqing Med Univ, Childrens Hosp, Dept Pediat Urol Surg, Chongqing 400014, Peoples R China"

通信作者："Shi, QL (通讯作者)，Chongqing Med Univ, Childrens Hosp, Key Lab Child Dev & Disorders, Minist Educ,Chongqing Key Lab Pediat,Pediat Res In, Chongqing 400014, Peoples R China.; Shi, QL (通讯作者)，Chongqing Med Univ, Childrens Hosp, Dept Pediat Urol Surg, Chongqing 400014, Peoples R China."

来源：JOURNAL OF ONCOLOGY

ESI学科分类：CLINICAL MEDICINE

WOS号：WOS:000868325800004

JCR分区：Q2

影响因子：4.501

年份：2022

卷号：2022

期号：　

开始页：　

结束页：　

文献类型：Article

关键词：　

摘要："Objective. The dysfunction of immune surveillance, a hot spot in cancer research, could lead to the occurrence and development in multicancers. However, the potential mechanisms of immunity in Wilms' tumor (WT) remain unclear on Wilms' tumor (WT). In this study, we aim to investigate the immune cell in WT and explore the underlying treatment strategy. Method. We quantified stromal and immune scores by using ESTIMATE algorithm based on gene expression matrix of WT patients in TCGA and GEO databases. Different expression genes (DEGs) and functional enrichments were analyzed by R studio and DAVID tools. Flow cytometry, immunofluorescence staining, ELISA assay, and qRT-PCR were used for detecting the NK cells, cytotoxic cytokines (INF-gamma, PRF, and GZMB), and NK cell receptor expression, respectively. WT patient autologous NK cells were stimulated by IL-2 and IL-15, and the cytotoxicity of NK cells against WT cell lines was detected by LDH assay. Western blot experiment was used for measuring the MAPK signaling pathway protein maker in NK cells. Results. ESTIMATE indicated that WT tissue had a lower immune score than adjacent kidney tissue. Meanwhile, the low immune score group was associated with poorly outcomes. DEG functional enrichment analysis showed that NK cell-mediated cytotoxicity was significantly different in low and high immune score groups. Although few of proportion of NK cells in WT patients were increased, most of that were significantly lower than normal children. Moreover, the proportion of NK cells and the expression level of INF-gamma, PRF, and GZMB in WT tissue were lower than adjacent kidney tissue. Importantly, the NKG2D expression level of NK cells was significantly lower in WT tissue. Furthermore, in vitro, compared with uncultured NK cells, IL-2 and IL-15 could effectively enhance the cytotoxicity of NK cells on killing the WT cell lines. The FACS and WB results showed that the NKG2D and p-PI3K ratio PI3K, MEK1/2, and p-ERK1/2 ratio ERK1/2 were significantly increased in IL-2 and IL15 group compared with uncultured groups. Conclusion. The abnormal NK cell-mediated cytotoxicity may cause the occurrence of WT. Costimulation of WT patients autologous NK cells could effectively enhance the antitumor reaction which involved in activation of NKG2D-mediated MAPK signaling pathway."

基金机构："Natural Science Foundation of Chongqing, China [cstc2021jcyj-msxm-X0549]; Postdoctoral Research Project of Chongqing, China; National Clinic Key Specialty Project , China [2013(544)]; Program for Youth Innovation in Future Medicine, Chongqing Medical University , China [W0069]"

基金资助正文："The authors would like to thank the urology team for supporting this work. This work was supported by the Natural Science Foundation of Chongqing, China (No. cstc2021jcyj-msxm-X0549); Postdoctoral Research Project of Chongqing, China; National Clinic Key Specialty Project (2013(544)), China; and Program for Youth Innovation in Future Medicine, Chongqing Medical University (W0069), China."