Abstract
ObjectiveTo investigate the effects of CCL21/CCR7 on the proliferation, migration, and invasion of T24 cells and the possible associated mechanisms: expression of MMP-2 and MMP-9, and regulation of BCL-2 and BAX proteins.MethodsT24 cells received corresponding treatments including vehicle control, antibody (20ng/mL CCR7 antibody and 50 ng/ml CCL21), and 50, 100, and 200 ng/ml CCL21. Proliferation was evaluated by MTT assay; cell migration and invasion were assayed using a transwell chamber. Cell apoptosis was induced by Adriamycin (ADM). The rate of cell apoptosis was examined by flow cytometry using annexin V-FITC/PI staining. Western-blot was used to analyze MMP-2 and MMP-9 and BCL-2 and BAX proteins.ResultsCCL21 promoted T24 cell proliferation in concentration-dependent manner with that 200 ng/mL induced the largest amount of proliferation. Significant differences of cell migration were found between CCL21treatment groups and the control group in both the migration and invasion studies (P < 0.001 for all). The expressions of MMP-2 and MMP-9 proteins were significantly increased after CCL21 treatment (p < 0.05 for all). Protein expression of Bcl-21 follows an ascending trend while the expression of Bax follows a descending trend as the concentration of CCL21 increases. No difference was found between the control group and antibody group for all assessments.ConclusionCCL21/CCR7 promoted T24 cell proliferation and enhanced its migration and invasion via the increased expression of MMP-2 and MMP-9. CCL21/CCR7 had antiapoptotic activities on T24 cells via regulation of Bcl-2 and Bax proteins. CCL21/CCR7 may promote bladder cancer development and metastasis.
Highlights
Bladder cancer is one of the most common types of adult cancer
The expressions of MMP-2 and matrix metalloproteinase9 (MMP-9) proteins were significantly increased after CCL21 treatment (p < 0.05 for all)
Protein expression of Bcl-21 follows an ascending trend while the expression of Bax follows a descending trend as the concentration of CCL21 increases
Summary
Bladder cancer is one of the most common types of adult cancer. In 2008 alone, 386,000 patients were diagnosed with bladder cancer which resulted in 150,200 deaths worldwide based on global statistics[1]. Chemokines, a superfamily of small secreted peptides characterized by their ability to induce leukocyte migration, together with their receptors have been found to be involved in the migration of cells of the lymphoid system, may affect cancer development and progression[2,3,4]. [6] CCR7 is expressed by various types of lymphocytes including naive B and T cells, semimature and mature dendritic cells, and T regulatory cells[5]. CCR7 and its ligand (s) may participate in the proliferation, progression, and metastasis of cancer cells of various organ origins [7,8,9,10,11,12,13]. We found that CCR7 was involved in the development and progression of bladder cancer (unpublished data)
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