Yang, D. L.;Xin, M. M.;Wang, J. S.;Xu, H. Y.;Huo, Q.;Tang, Z. R.;Wang, H. F.
Genetics and molecular research : GMR,2015年14(4):17699-17707 ISSN：1676-5680
[Xin, M. M.; Huo, Q.; Wang, H. F.; Xu, H. Y.; Wang, J. S.; Yang, D. L.; Tang, Z. R.] Department of Urology, The Second Affiliated Hospital of Kunming Medical University, Yunnan Institute of Urology, Kunming, China
[Wang, HF] Kunming Med Univ, Yunnan Inst Urol, Affiliated Hosp 2, Dept Urol, Kunming, Peoples R China.
It is well known that chemokine receptors and their ligands play important roles in mediating the invasion and metastasis of malignant tumors. This aim of this study was to investigate the expression and clinical significance of chemokine receptor CXCR4 and its ligand CXCL12 in bladder tumor tissues. Cancerous and adjacent normal bladder tissues were collected from 42 patients. The expressions of CXCR4 and CXCL12 proteins were then detected by immunohistochemistry, and the expressions of CXCR4 and CXCL12 mRNAs were detected by RT-PCR. Bladder cancer tissues showed higher positive expressions of CXCR4 and CXCL12 than those in normal bladder mucosal tissues (z = 7.332, 6.758, P < 0.001). Positive expressions of CXCR4 and CXCL12 were related to the differentiation degree and invasive depth of cancer tissues (z = 2.598-4.594, P < 0.05), but not to patient gender or age (z = 0.273-0.554, P > 0.05). The expression of CXCR4 was positively correlated to CXCL12 expression in bladder cancer tissues (r = 0.661, P < 0.05). RT-PCR revealed that CXCR4 and CXCL12 mRNAs were not expressed in normal tissues. Moreover, with increased depth of invasion, CXCR4 and CXCL12 mRNA expressions gradually increased in bladder cancer tissues and showed significant intergroup differences (F = 56.642, 67.928, P < 0.01). Taken together, these results indicate that the chemokine receptor CXCR4 and its ligand CXCL12 play important roles in the occurrence and development of bladder cancer.
[Wang, HF] Kunming Med Univ, Dept Urol, Affiliated Hosp 2, Yunnan Inst Urol, Kunming, Peoples R China.
Urinary bladder;implantation metastasis;animal model;green fluorescent protein (GFP)
OBJECTIVE: To establish a fluorescent implantation metastasis model of bladder carcinoma with high metastatic potential in nude mice and observe development and metastasis. METHODS: Human bladder cancer EJ cells with high invasive ability were screened and transfected with GFP plasmid to screen stable enhanced GFP-expressing clones instilled into the bladders of nude mice. Subsequent growth, invasion, and metastasis of the implanted tumors were observed and evaluated with a whole-body fluorescence optical imaging system. RESULTS: The transfected bladder cancer EJ cells stably and efficiently expressed EGFP. The growth, invasion and metastasis of the implant bladder tumor were readily observed and accurately evaluated by fluorescent microscopy. In the bladders of nude mice, the rates of EGFP expression detected by flow cytometry at weeks 1-4 were 22.6%, 46.7%, 62.3% and 72.7%, respectively, with clear increase over time. CONCLUSION: GFP-labeled bladder cancer EJ cells display green fluorescence under fluorescent microscopy and show stable GFP expression. The model will provide a simple and reliable means for studying the mechanism of implantation metastasis of human bladder cancers in vivo.
Background: Metastasis of tumor implantation includes a series of processes from detachment from the primary tumor to formation of the implanted metastase. Tumor cells survival in urine is a necessary condition for metastasis. Adaptation to urine is essential for this. Objective: Establish a urine-resistant cell sub-strain of human bladder cancer cell line (ET cell lines), and study different characteristics compared to parent cells. Methods: EJ cell lines were cultured in nutrient medium. Urine-resistance cell sub-strain (EJ-U) was harvested after prolonged culture by gradually increasing the concentration of urine. Gen chip was used to detect the genome series of EJ and EJ-U and to analyze the difference of gene expression. Results: EJ-U in urine had a higher survival rate after 24 hours in urine compared with El The EJ-U had almost the same growth velocity with EJ, and they had the analogous growth curves. The time-duration for EJ-U to survive was longer than EJ in urine. In gene ontology analysis, 272 significant different genes were found. Conclusion: EJ-U cell sub-strain was more adaptable than its parent cell lines EJ. The different genes may explain the reason why bladder cancer cells could survive for a long time in urine.
We investigated the effect of membrane toxin 12 (MT-12), isolated from Naja naja atra, a species of Chinese Cobra, on the proliferation and invasion of human bladder cancer EJ cells and studied its mechanisms using MTT assay, Transwell chamber invasion assay, scanning electron microscopy and flow cytometry. The results indicated that MT-12 inhibited the proliferation of bladder cancer cells in a dose-dependent manner. The half maximal inhibitory concentration (IC50) after 72 h was 0.66 microg/ml. The invasion of cells decreased with increasing doses of MT-12 (0.125-0.5 microg/ml, P<0.001). Expression of CXCR4 decreased with the effect of MT-12 for a particular concentration range (0.125-0.5 microg/ml). To conclude, MT-12 is capable of inhibiting the proliferation and invasion of bladder cancer EJ cells. This mechanism may be associated with reduced expression of CXCR4 protein.
Objective: To evaluate clinical effect of the ileal neobladder after modified radical cystectomy and standard radical cystectomy. Clinical Materials: Since June, 1991 to June, 2008, 262 consecutive cases with bladder carcinoma were treated by radical cystectomy and bladder