|Table of Contents|

Tumor-specific effective transform growth factor-β-insensitive T cells inhibit the proliferation and metastasis of renal cancer stem cells

Journal Of Modern Oncology[ISSN:1672-4992/CN:61-1415/R]

Issue:
2025 03
Page:
359-366
Research Field:
Publishing date:

Info

Title:
Tumor-specific effective transform growth factor-β-insensitive T cells inhibit the proliferation and metastasis of renal cancer stem cells
Author(s):
LIU Youhuang1; TIAN Feng2; ZHANG Chao3; YUE Dezhou3; WANG Longxin3
1.Department of Urology,Eastern Theater Command General Hospital,Jiangsu Nanjing 210002,China;2.Department of Urology,Affiliated Zhongshan Hospital of Dalian University,Liaoning Dalian 116000,China;3.Department of Urology,Tianyinshan Hospital,the First Affiliated Hospital of China Pharmaceutical University,Jiangsu Nanjing 211100,China.
Keywords:
transforming growth factor-β; renal cell carcinoma; CD8+ T cells; cancer stem cells; epithelial-mesenchymal transition
PACS:
R737.11
DOI:
10.3969/j.issn.1672-4992.2025.03.002
Abstract:
Objective:To investigate the tumor-specific,effective transform growth factor-β (TGF-β)-insensitive CD8+ T cells antitumor effects for CD105+ renal cell carcinoma stem cells in vitro and vivo.Methods:In the present study,we isolated tumor-reactive CD8+ T cells from RCC patients,activated them with autologous immature dendritic cells (DC) and freeze -thraw tumor lysate,rendered them insensitive to TGF-β by infection with a containing dominant-negative TGF-β-Ⅱ receptor (TβRⅡDN) retrovirus,the immunophenotype of tumor-specific transform growth factor-β-insensitive CD8+ T cells (TβRⅡDN-CTL ) were analyzed by flowcytometry and subsequently tested their antitumor responses both in vitro and in vivo.Isolated CD105+ cells from the primary renal cell carcinoma cell lines using immunomicrobeads and the cancer stem cell markers such as Vimentin，Pax2,Oct4,Nanog were analyzed.The effect of TGF-β1 induction on CD105+ cells was detected in vitro,and the killing effect of TβRⅡDN-CTL and Nave CTL on CD105+ cells in vitro was observed by 51Cr release experiment.The tumor growth of the mice among the three groups was observed,and the lung metastasis,tumor apoptosis and lymphocyte infiltration were measured.Furthermore,the EMT-related molecules were analyzed by RT-PCR and westen-blotting in the tumor tissues.Results:The Vimentin,Pax2,Oct4,Nanog were high expressed in CD105+ cells isolated from primary renal cancer cell lines whereas the vWF,CK were low expressed by immunofluorescence and western-blotting analysis.In the TGF-β1 induced condition,CD105+ cells showed the decreased expression of E-cadherin and high expression of N-cadherin and F-actin,which indicated EMT effects were existed.In the TβRⅡDN-CTL cells,the dominant phenotype was the CD27+CD45 RA- which is phenotype of effector CD8+ T cells whereas the Nave CD8+ T cells were identified as CD27+CD45 RA+,which is consistent with unprimed CD8+ T cells（P＜0.05） analyzed by flow cytometry.Western-blotting experiments showed the levels of phosphorylated Smad-2 protein in TβRⅡDN-CTL were undetectable which showed its TGF-β insensitive property.The 51Cr release assay showed the TβRⅡDN -CTL cells exhibited a 5-fold higher tumor-killing activity for CD105+ cells compared with Nave CD8+ T cells（P＜0.01）.In vivo experiments,the TβRⅡDN CTL group showed the lowest tumor volume and weight,the fewest pulmonary metastases,high tumor cell apoptosis,more CD8+T lymphocyte infiltration as compared with other groups.Further experiments showed that the EMT related molecules were inhibited in tumor tissues.Conclusion:The novel TGF-β-insensitive tumor-specific effector CD8+ T cells have strong specific anti-tumor effects on CD105+ cells in vitro,and can also show strong anti-tumor activity in vivo.This immunotherapy strategy may offer the promise of a novel therapeutic option for the treatment of renal cell carcinoma.

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Memo

Memo:
National Natural Science Foundation of China(No.81372741，81472392）；国家自然科学基金资助项目（编号：81372741，81472392）
Last Update: 1900-01-01