Mechanism of Telomerase Reverse Transcriptase Regulating Dendritic Cell Immune Function in Proliferation and Cell Cycle of Renal Cell Carcinoma

Abstract

To select human renal cell carcinoma cell lines VMRC-RCZ After resuscitation, culture and passage, the cell concentration was adjusted to 1 × 106/ml, and the experiment was divided into a control group, a renal cancer cell + DC group (DC group), and a renal cancer cell + DC+ hTERTtransfection group (transfection Group), a total of 3 groups; the recombinant eukaryotic expression plasmid hTERT-IRES2-EGFP was constructed and transfected into mature DC; the renal cancer cell + DC group was transfected with the empty plasmid, and the renal cancer cell + DC+ hTERT transfection group was transfected with the overexpression plasmid The transfection efficiency was detected by RT-PCR method. Cultured for 12 h and 24 h respectively, the percentage of HLA-DR, CD40 and MHC-II molecules in three groups of DC immunophenotypes was determined according to flow cytometry assay, and the cytokines IL-12 and TNF-α expression level, DC apoptosis rate was detected by in situ hybridization, the rate of tumor cell proliferation was measured according to MTT method, and cell cycle was determined through flow cytometry.