journal contribution
posted on 2023-03-31, 19:44 authored by Jiangyong Yu, Shuhang Wang, Wei Zhao, Jianchun Duan, Zhijie Wang, Hanxiao Chen, Yanhua Tian, Di Wang, Jun Zhao, Tongtong An, Hua Bai, Meina Wu, Jie Wang <p>Supplemental Experimental Procedures The establishment of PDX models. Table S1. PDXs and corresponding clinical data Table S2: Antibody for Flow Cytometry. Table S3: qRT-PCR primer sequences. Table S4: Antibodies used in Western blots. Supplemental Experimental data Figure S1: Stem cell properties of α2Î'1+ H69 cells. Figure S2: The proportion of É'2Î'1+ and CD133+ cells in parental and 2nd generation PDXs Figure S3: Treatment curves of five PDX models Figure S4:Stem cell properties of α2Î'1+ Bclu80 cells. Figure S5: Differential genes and pathway enrichment in É'2Î'1 positive H1048 cells by RNA-Sequencing and analysis of protein interaction network. Table S5: Comparison of tumorigenic sufficiency for É'2Î'1/CD133 FACS-sorted H1048 cells in vivo. Table S6: Comparison of tumorigenic sufficiency for É'2Î'1 FACS-sorted H1048, H69 cells and Bclu80 PDX models in vivo. Table S7: Serial transplantation assay was performed into mice for validating self-renewal in vivo. Table S8: HE staining and immunochemistry results of SCLC patients and corresponding PDX models. Table S9. The differential expression genes of É'2Î'1+ and É'2Î'1- H1048 cells using RNA-Sequencing. Supplemental References</p>
Funding
Ministry of Education
Central Laboratory, Key Laboratory of Carcinogenesis and Translational Research, Ministry of Education, Beijing Cancer Hospital & Institute
Mayo Clinic
History
ARTICLE ABSTRACT
Purpose: Chemoresistance in small-cell lung cancer (SCLC) is reportedly attributed to the existence of resistant cancer stem cells (CSC). Studies involving CSC-specific markers and related mechanisms in SCLC remain limited. This study explored the role of the voltage-dependent calcium channel α2δ1 subunit as a CSC marker in chemoresistance of SCLC, and explored the potential mechanisms of α2δ1-mediated chemoresistance and strategies of overcoming the resistance.Experimental Design: α2δ1-positive cells were identified and isolated from SCLC cell lines and patient-derived xenograft (PDX) models, and CSC-like properties were subsequently verified. Transcriptome sequencing and Western blotting were carried out to identify pathways involved in α2δ1-mediated chemoresistance in SCLC. In addition, possible interventions to overcome α2δ1-mediated chemoresistance were examined.Results: Different proportions of α2δ1+ cells were identified in SCLC cell lines and PDX models. α2δ1+ cells exhibited CSC-like properties (self-renewal, tumorigenic, differentiation potential, and high expression of genes related to CSCs and drug resistance). Chemotherapy induced the enrichment of α2δ1+ cells instead of CD133+ cells in PDXs, and an increased proportion of α2δ1+ cells corresponded to increased chemoresistance. Activation and overexpression of ERK in the α2δ1-positive H1048 cell line was identified at the protein level. mAb 1B50-1 was observed to improve the efficacy of chemotherapy and delay relapse as maintenance therapy in PDX models.Conclusions: SCLC cells expressing α2δ1 demonstrated CSC-like properties, and may contribute to chemoresistance. ERK may play a key role in α2δ1-mediated chemoresistance. mAb 1B50-1 may serve as a potential anti-SCLC drug. Clin Cancer Res; 24(9); 2148–58. ©2018 AACR.
