Activation of the transcription factor E2F-1 gene is a negative event in dendritic cell (DC) maturation process. Down-regulation of E2F1 causes immaturity of DC thereby stopping antigen production which in turn leads to inhibition of immune responses. E2F-1-free stimulates the NF-kB signaling pathway, leading to activation of monocytes and several other transcription factor genes. In the study, we report that down-regulation of E2F-1 in DCs promote anti-tumor immune response in gastric cancer (GC) cells through a novel mechanism. DCs were isolated from peripheral blood mononuclear cells. E2F-1 small interfering RNA (E2F-1-shRNA) induced down-regulation of E2F-1 mRNA and protein expression in DCs. Furthermore, we identified the E2F-1-shRNA targeted the CD80, CD83, CD86, and MHC II molecules, promoted their expression, and induced T lymphocytes proliferation activity and up-regulation of IFN-Ī³ production and GC cell killing effect, which significantly correlated with the cytotoxic T lymphocytes activated by E2F-1-shRNA DCs. The higher expression of miR-34a was found which was significantly correlated with the DC enhancing anti-tumor immunity against gastric cancer cell, and miR-34a potently targeted DAPK2 and Sp1, both of which were involved in the deactivation of E2F-1. Moreover, in E2F-1-DC-down-regulation in mice, GC transplantation tumors displayed down-regulation of Sp1, DAPK2, Caspase3, and Caspase7 and progressed to anti-tumor immunity. Collectively, our data uncover an E2F-1-mediated mechanism for the control of DC anti-tumor immunity via miR-34a-dependent down-regulation of E2F-1 expression and suggest its contribution to GC immunotherapy.
Transcription factor E2F-1 Dendritic cell MicroRNA Gastric cancer cells Immunotherapy
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We acknowledge financial support from the National Science Foundation for Young Scholars of China (No. 81502120), the Science Foundation for Young Scholars of Guangxi Medical University (No. GXMUYSF201404), and the Scientific Research and Technological Foundation of Guangxi (No.1140003A-35). Also, we express our gratitude to Ms. Shwetha Manoj, who made extraordinary help in paper editing.
Compliance with ethical standards
The study design was approved by the Guangxi medical university ethic review board obeying the Helsinki Declaration. All participants have signed informed consent forms and agreed the treatment.
The mice were normatively managed by SPF laboratory in the Animal Central of Guangxi Medical University, following the rules of ethical regulations for medical experimental animal care.
Conflicts of interest
The authors declare that there is no conflict of interests regarding the publication of this paper.
Supplementary figure 1Down-regulation of E2F-1 decreased miR-23a, miR-27a, miR-126, miR-133a, and miR-148b, and increased miR-34a expression. The determination of miRNA expression levels of miR-23a, miR-27a, miR-126, miR-133a, miR-148b, and miR-34a through semiquantitative reverse-transcriptase polymerase chain reaction. (DOCX 170Ā kb)
Supplementary figure 2FACS analysis of peripheral blood monocyte-derived DCs. PBMCs cultured in the presence of GM-CSF (50Ā ng/mL), IL-4 (50Ā ng/mL), and LPS (100Ā ng/mL). Maturation of DCs is marked by the increased levels of CD83 and CD86. There is also a marked increase in the expression of HLA-DR. The numbers in the boxes indicate the mean fluorescence intensity. (DOCX 270Ā kb)
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