Adenovirus-mediated TK gene combined with α-IFN in treatment of nude mice transplanted with human renal clear-cell carcinoma / 第二军医大学学报

Objective: To investigate the inhibitory effect of adenovirus-mediated herpes simplex virus-thymidine kinase (TK) gene combined with α-IFN on renal clear-cell carcinoma. Methods: Adenovirus containing suicide gene TK, in combination with GCV or α-IFN, was used to treat human renal clear-cell carcinoma cell line 786-0, and the in vitro cytotoxic effects against 786-0 were evaluated using MTT method. The subcutaneous transplantation model of 786-0 cells was established with nude mice. Adenovirus containing TK gene was injected intratumorally and the GCV (50 mg/kg) was injected intraperitoneally; α-IFN (104 U/L) was injected intratumorally in combined therapy. The growth of tumors was observed after treatments. Results: The survival rate of 786-0 cells was (35.07 ± 1.43)% in the TK+GCV+α-IFN group, (68.57 ± 1.41)% in the TK+ GCV group and (68.65 ± 1.45)% in the α-IFN group (P=0.000). There was an obvious synergic effect between Ad-TK and α-IFN in inhibiting 786-0 cells. Ad-TK combined with GCV and α-IFN significantly suppressed the growth of 786-0 cells growth in nude mice model. Conclusion: Adenovirus-mediated TK plus prodrug GCV combined with α-IFN has obvious therapeutic effect in treatment of human renal clear-cell carcinoma.

Detection of Skp2 and p27kip1 expression in human renal cell carcinoma using tissue chip technique / 南方医科大学学报

<p><b>OBJECTIVE</b>To detect the expression of skp2 and p27kip1 in human renal cell carcinoma (RCC) using tissue chip technique, and evaluate the relationship between the proteins and the biological behavior of RCC.</p><p><b>METHODS</b>Tissue chip technique and immunohistochemical SP method was used to detect the expression of skp2 and p27kip1 in normal and tumor tissues.</p><p><b>RESULTS</b>The positivity rate of Skp2 in RCC was significantly higher than that in normal renal tissues (P=0.025). The positivity rate of Skp2 expression in RCC was significantly correlated to poor differentiation of the tumor (P=0.002), and was not associated with the patients gender, age, tumor size, lymph node metastasis and stages of RCC (P>0.05). The positivity rate of p27kip1 in RCC was significantly lower than that in normal renal tissues (P=0.007). The positivity rate of p27kip1 expression was inversely correlated to the malignancy and stage of RCC (P<0.05), but not with the patients' age, gender, lymph node metastasis and tumor size (P>0.05). An inverse correlation was noted between Skp2 and p27kip1 expressions (r= -0.273, P=0.014).</p><p><b>CONCLUSION</b>Overexpression of Skp2 protein may lead to decreased p27kip1 level in RCC, indicating its involvement in the carcinogenesis and development of RCC.</p>

Whole-body fluorescent imaging of the growth and metastasis of GFP-expressing bladder tumors / 南方医科大学学报

<p><b>OBJECTIVE</b>To label a human bladder cancer cell line and establish a novel human bladder cancer mouse model.</p><p><b>METHODS</b>T-24 cells, a human bladder transitional cell carcinoma cell line, were transfected with GFP plasmid to screen stable GFP-expressing clones. The latter were implanted into the wall of the bladder or the subcutaneous tissue of the neck of nude mice. The growth, invasion, and metastasis of the implanted tumor were observed and evaluated with whole-body optical imaging system. The findings were compared with those of HE staining on routine paraffin sections.</p><p><b>RESULTS</b>GFP-labeled tumor cells displayed green fluorescence under fluorescent microscopy and showed stable GFP expression in vitro and in vivo. One week after in situ transplantation of 5 x 10(5) T24 cells, the new bladder cancer was observed and evaluated under whole-body optical imaging system. Two weeks later, the new bladder tumor could be palpated, and 4 weeks later, metastasis to regional drainage lymph nodes in the pelvic and retroperitoneal lymph nodes occurred. The growth and metastasis of the implant bladder tumor were easily observed and accurately evaluated by fluorescent microscope.</p><p><b>CONCLUSION</b>GFP-labeled tumor cells display green fluorescence under fluorescent microscopy and show stable GFP expression. GFP-labeled T-24 cells and the novel human bladder cancer model described hereby provide a simple and reliable means for studying human bladder cancer in vivo.</p>

Cytotoxic effect of oncolytic virus combined with mitomycin against human bladder cancer cells in vitro and in vivo / 南方医科大学学报

<p><b>OBJECTIVE</b>To evaluate the effect of combined use of oncolytic virus and the chemotherapeutic agents mitomycin (MMC) in growth inhibition of human bladder cancer cell line T-24 in vitro.</p><p><b>METHODS</b>Human bladder cancer cell line T-24 was infected with oncolytic virus (ONYX-015) of different multiplicity of infection, or treated with MMC in addition to ONYX-015. The changes in the cell growth, morphology, and apoptosis of cultured T-24 cells were observed by means of cell counting and fluorescence microscopy after the treatments. The effects of the treatment protocols were also tested in nude mouse model of implanted subcutaneous tumor.</p><p><b>RESULTS</b>Combined use of ONYX-015 and MMC produced substantially stronger cytotoxic effect against T-24 cells than exclusive use of ONYX-015. In in vivo experiments, combination of oncolytic virus and MMC resulted in much more significant tumor growth inhibition than either of the agents used alone. Obvious T-24 cell apoptosis could be observed in response to combined ONYX-105 and MMC treatment and exclusive ONYX-105 treatment.</p><p><b>CONCLUSIONS</b>ONYX-015 combined with MMC can produce significant cytotoxicity against T-24 cells and enhance therapeutic efficacy against bladder carcinoma.</p>

Expression of TFAR19(PDCD5) in normal human kidney, renal clear cell carcinoma, normal human bladder and bladder carcinoma / 南方医科大学学报

<p><b>OBJECTIVE</b>To detect the expression of apoptosis gene PDCD5 in tissues of normal human kidney, renal clear cell carcinoma, normal bladder and bladder carcinoma, and explore the role of PDCD5 gene in renal clear cell carcinoma and bladder carcinoma.</p><p><b>METHODS</b>Indirect immunohistochemistry was employed to detect PDCD5 expression in 63 kidney specimens and 42 bladder specimens. Positive expression rates and intensity of PDCD5 protein expression in the kidney tissue were investigated microscopically and by computerized image analysis. Positive expression rate in the bladder tissue was investigated by microscopic observation.</p><p><b>RESULTS</b>The results of immunohistochemical staining showed PDCD5 protein overexpression in the renal tubule of normal human kidney tissues and downregulation with the stage increase of renal clear cell carcinoma. PDCD5 protein expression showed statistical significance in tissues of normal kidney and renal clear cell carcinoma in all stages. No obvious PDCD5 expression was detected in the tissues of normal human bladder and bladder carcinoma.</p><p><b>CONCLUSION</b>PDCD5 is an important apoptosis-regulating factor in the occurrence of renal clear cell carcinoma, and its expression is extremely low in tissues of normal human bladder and bladder carcinoma.</p>