RelB Research Articles

Correction: S100A9+ MDSC and TAM-mediated EGFR-TKI resistance in lung adenocarcinoma: the role of RELB.

[This corrects the article DOI: 10.18632/oncotarget.24146.].

RelB-siRNA enhanced the radiosensitivity of murine prostate cancer cell line RM-1

To explore the transfecting effects of RelB small interfering RNA (RelB-siRNA) on murine prostate cancer cell line RM-1 sensitivity of radiotherapy. The RM-1 cells were divided into RelB-siRNA vector, empty vector, non-transfection and normal groups. After transfection, the first three groups were irradiated by X ray. Clone formation array method was used to measure the surviving fraction and the relevant parameter values after 0, 2, 4, 6, 8 Gy irradiation. Apoptotic rate was measured via Annexin V/PI flow cytometry after 6 Gy irradiation. The level of RelBmRNA was estimated by real-time polymerase chain reaction (PCR) after 6 Gy irradiation. And the RelB protein of cytoplasm and nucleus was detected by Western blot after 6 Gy irradiation. Clone formation array showed that RelB group at each dose point corresponding to the survival fraction were lower that the empty vector nd non-transfection groups. The D0, Dq and SF2 values of RelB group were 1.68, 0.60, 0.43, lower than those of empty vector group 1.92, 3.08, 0.89 and non-transfection group 1.93, 2.76, 0.84. Annexin V/PI flow cytometry demonstrated that the apoptotic rate of RelB group was 15.27% ± 1.62% and it was significantly higher than the non-transfection group 7.90% ± 1.50% and the empty vector group 8.40% ± 0.69% respectively (P < 0.01) .Real-time PCR indicated that RelB-mRNA amplification of RelB group was 1.18 ± 0.03 and it was significantly lower than the non-transfection group 2.10 ± 0.61 and the empty vector group 1.97 ± 0.66 respectively (P < 0.01) .Western blot suggested that cytoplasmic gray-scale ratio of RelB group was 0.50 ± 0.08 and it was significantly lower than the non-transfection 1.77 ± 0.19 and the empty group 1.52 ± 0.12 respectively (P < 0.01) . And the nuclear gray-scale radio of the RelB group was 0.18 ± 0.03 and it was significantly lower than the non-transfection group 0.61 ± 0.12 and the empty group 0.54 ± 0.13 respectively (P < 0.01) . The RelB protein inhibition rates of cytoplasmic and nuclear RelB-siRNA were 71.8% and 70.4% respectively. RelB-siRNA can effectively inhibit the RelB expression of murine prostate cancer cell line RM-1 and significantly increase its sensitivity to radiotherapy.

Lentiviral-mediated short hairpin RNA against RelB produces the generation of tolerogenic dendritic cells

Objective Lentiviral-mediated short hairpin RNA (shRNA) against RelB was used to produce tolerogenic dendritic cells from murine bone marrow derived dendritic cells (BMDCs).Methods RelB expression in the BMDCs was silenced by Lentivirus carrying RelB shRNA.The RelB expression was detected by reverse transcriptase-polymerase chain reaction (RT-PCR) and Western blotting.The apoptosis rate and surface markers of dendritic cells (DCs) were assessed by flow cytometry.Interleukin (IL)-12,IL-10,transforming growth factor-β1 (TGF-β1) secreted by DCs and DNA binding capacity of nuclear factor-κB (NF-κB) subunits in the nucleus were measured by enzyme linked immunosorbent assay (ELISA),independently.Results RelB expression was significantly inhibited in DCs following lentiviral mediated delivery of RelB specific shRNA.The RelB shRNA-DC produced lower IL-12 [(96.3 ± 32.8) ng/L] and higher IL-10 [(218.2 ± 43.3) ng/L] than mature dendritic cells (mDCs) and silencing control DCs.There were no difference in the apoptosis rate between shRNA ReIB-DCs and mDCs.The expression levels of co-stimulatory molecules MHC-Ⅱ (0.34 ± 0.04),CD80 (0.38 ± 0.03),CD86 (0.17 ± 0.02),CD83 (0.21 ± 0.02)were lower in the RelB shRNA-DCs than in the mDCs and silencing control DCs.In addition,RelB shRNA also inhibited the RelB DNA binding capacity (0.28 ±0.06) but had no effect on otherNF-κB subunits.Conclusion RelB shRNA transfection of DCs can induce the immature status,and produce tolerogenic DCs. Key words: RelB ; Dendritic cell; RNA interference ; Gene silencing

Prevention of acute rejection by RelB siRNA-silenced dendritic cells

Objective To prevente acute rejection in cardiac transplantation by using tolerogenic dendritic cells (DC).Methods The mixed lymphocyte reaction (MLR) was performed by RelB siRNA-DC with allogenic antigen-specific T cells.The apoptosis rate of T cells and the ratio of CD4 + CD25 + Foxp3 + regulatory T cells were detected by using flow cytometry after MLR.In the murine model of cardiac transplantation,the survival time was observed after infusion of RelB siRNA-DC before transplantation.HE staining was used to examine the rejection degree in the specimens of allografts.Results Mature DC and LucR siRNA DC could stimulate the proliferation of T cells more strongly than immature DC and RelB siRNA-DC (P ＜ 0.05).The RelB siRNA-DC could induce allogeneic T cell hyporesponsiveness,but had no specific reactivity on the third kind T cells (P ＞ 0.05).RelB siRNA-DC induced T cell apoptosis [(29.21 ± 7.55)％,P ＜ 0.05] and promoted T cells differentation into CD4 + CD25 + Foxp3 + regulatory T cells [(65.3 ± 12.6)％,P ＜ 0.05].In the mouse cardiac transplantation model,infusion of donor-derived Lenti-RelB siRNA-DC could prolong the graft survival time (P ＜ 0.05) and reduce rejection occurrence.Conclusion Donor-derived RelB-siRNA-induced tolerogenic DC can significantly induce immune tolerance in vitro and in vivo. Key words: Dendritic cell; Heart transplantation; Rejection; Regulatory T cell

Construction and identification of lentiviral vector of RNA interference of RelB gene

Objective To construct a recombinant lentivirus plasmid of RNA interference (RNAi)of RelB gene.Methods According to the GenBank information of RelB,5 complementary DNA containing both sense and antisense Oligo DNA of the targeting sequences were designed,synthesized,and cloned into PlentiLox 3.7 which contained U6 promoter and green fluorescent protein(GFP).The positive clones were confirmed by PCR,sequencing,and authentication with KspA Ⅰand Hind Ⅲ.RM-1 cells were transfected and detected by Western-blot.293T cells were cotransfected with lentiviral vector and virus packaging plasmids.All virus stocks were produced by calcium phosphate mediated transfection.Results PCR and DNA sequencing demonstrated that the lentivirus RNAi vector of RelB was constructed successfully.The most effective siRNA sequence was 5'-TTGGAAATCATCGACGAAT-3'.The titer of concentrat ed virus was 8×1010 pfu/L.Conclusion The lentivirus RNAi vector of RelB was constructed successfully.The lentivirus RNAi vector of RelB significantly inhibited the RelB expression. Key words: siRNA; Lentiviral vector; Construction; Identification

Weight, food intake, and body composition: effects of exercise and of protein deficiency.

Weight, food intake, and body composition: effects of exercise and of protein deficiency.