ABSTRACT
@#Objective: To investigate the role of CpG ODN (CpG oligodeoxynucleotide) adjuvant in enhancing the anti-bladder cancer response induced by MAGE-3 (melanoma antigen gene -3) antigen and its molecular mechanism. Methods: Mononuclear cells were isolated from HLA-A2 type peripheral blood of healthy donors by Ficoll method to prepare mature DC by conventional means. DC surface markers were detected by flow cytometry. MTT assay was used to detect the promotion effect of DCs sensitized by different means (MAGE-3, CpGODN, MAGE-3+CpG ODN, irrelevant control antigen) on the proliferation of T lymphocytes and the killing effect of CTL on BIU-87 tumor cells. The tumor mass of nude mice bearing BIU-87 bladder cell xenograft were examined on Day 7 and 11 after CpG ODN+MAGE-3 sensitized DC treatment. The expression of Bcl-2/Bax protein was detected by Western blotting while the proliferation level of xenograft cells was detected by MTT assay. Results: DCs sensitized by CpG ODN combined with MAGE-3 antigenic peptides could promote the proliferation of T lymphocytes and significantly enhance the killing effect of CTLon target BIU-87 cells (P< 0.05). Compared with other sensitized DCs, in vivo experiments showed that 7 and 11 days after treatment, both the tumor volume and weight were significantly reduced (all P<0.05), and the proliferation ability of xenograft tumor was decreased (P<0.05). Compared with other sensitization means, CpG ODN+MAGE-3 especially exhibited obvious inhibitive effect on tumor growth on Day 11, and significantly promoted the proliferation of splenic monocytes of tumor bearing mice (P<0.01); moreover, Bcl-2 expression in xenograft tissues significantly decreased（P<0.01）while Bax expression significantly increased（P<0.05 or P<0.01）on Day 3 after treatment. Conclusion: CpG ODN can promote the inhibitory effect of MAGE-3 sensitized DC on bladder cancer BIU-87 cells, which will provide experimental basis for clinical application of DC vaccine in bladder cancer treatment.
ABSTRACT
Objective To investigate the expression of p 120-catenin in human bladder cell line BIU-87 and T24.And to explore the effect of down-regulation of p120-catenin on proliferation, invasion and adhesion of BIU-87 cell line. Methods The study was from Aug .2010 to May.2011.Real-time RT-PCR and Western blot were used to examine the expression of p 120-catenin in human bladder cell line BIU-87 and T24.There were three study groups:non-transfected group , the group transfected with p 120-catenin siRNA and the group transfected with control siRNA .p120-catenin siRNA was used to decrease the expression of p120-catenin.MTT assay and colony formation assay were used to study the BIU-87 cell growth and cell pro-liferation.The invasion of BIU-87 cells was measured by Transwell chamber assay .Cell adhesion artificial re-constituted basement membrane assay was used to examine the change of BIU-87 cell adhesion capacity . Results In both BIU-87 and T24 cells there were the expressions of p 120-catenin.But the expression in BIU-87 cells (0.11±0.39) was more than that in T24 cells (0.48±0.17).The decreased of p120-catenin ex-pression could enhance the proliferation (182.7±13.4%) and the invasiveness (217.5±15.9) and decrease the adhesion capacity (57.3±6.4%) of BIU-87 cells. Conclusions There is higher expression level of p120-catenin in lower-grade malignant bladder cancer cells .The down-regulation of p120-catenin promotes the bladder cancer proliferation and invasiveness of bladder carcinoma cells , and inhibited the bladder canc-er cell adhesion .