Prognostic value of preoperative serum albumin in patients with non-muscle-invasive bladder cancer undergoing transurethral resection of bladder tumor / 南方医科大学学报

<p><b>OBJECTIVE</b>To assess the value of preoperative serum albumin level in predicting the survival of patients with non-muscle-invasive bladder cancer (NMIBC) undergoing transurethral resection of bladder tumor (TURBT).</p><p><b>METHODS</b>Two hundred and sixteen newly diagnosed patients with NMIBC who underwent TURBT between January, 2007 and April, 2012 were retrospectively analyzed. The patients were categorized into low albumin (<40 g/L) and normal albumin (≥40 g/L) groups. The patient survival was estimated using the Kaplan-Meier method, and univariate and multivariate Cox proportional analyses were used to determine the hazard ratios (HRs) for the overall survival (OS).</p><p><b>RESULTS</b>Of the patients with available data, 82 (39%) and 127 (61%) patients were classified into low albumin (<40 g/L) and normal albumin (≥40 g/L) groups, respectively. Kaplan-Meier analysis showed a significantly worse 5-year OS in low albumin group than in normal albumin group (P=0.017). In the multivariate Cox regression analysis, after adjusting for confounding variables, the preoperative albumin level remained as an independent predictor for 5-year OS (HR: 3.102, 95%CI: 1.200-8.020, P=0.020).</p><p><b>CONCLUSION</b>A low preoperative albumin level predicts a poor 5-year OS in patients with NMIBC who underwent TURBT. Preoperative serum albumin can be a good prognostic factor for predicting survival of the patients with NMIBC treated with TURBT.</p>

Infiltrating mast cells promote neuroendocrine differentiation and increase docetaxel resistance of prostate cancer cells by up-regulating p21 / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the effect of infiltrating mast cells on neuroendocrine differentiation (NED) and docetaxel sensitivity of prostate cancer (PCa) cells in vitro.</p><p><b>METHODS</b>Human PCa cell lines (LNCaP and C4-2) were co-cultured with human mast cell line (HMC-1) in Transwell chambers. Androgen receptor (AR) was silenced in C4-2 cells using sh-AR lentivirus, and p21 was knocked down and overexpressed by transfecting C4-2 cells with pLKO.1-sh-p21 and pCMV-p21, respectively. The morphological changes of LNCaP and C4-2 cells were observed. MTT assay and colony formation assay were used to assess the proliferation of LNCaP and C4-2 cells. CCK8 assay was used to detect the cell viability of C4-2 cells following docetaxel trreatment. RT-qPCR and Western blotting were performed to determine the mRNA and protein expressions of neuroendocrine markers, AR and p21 in the cells.</p><p><b>RESULTS</b>Co-culture with HMC-1 cells enhanced the neuroendocrine phenotypes, inhibited the proliferation and up-regulated the expression of p21 in LNCaP and C4-2 cells. P21 positively regulated NED through a non-AR-dependent signaling pathway, while p21 knockdown partially reversed NED promoted by the mast cells. PCa cells co-cultured with HMC-1 cells showed increased resistance to docetaxel, and silencing p21 partially reversed docetaxel resistance in PCa cells.</p><p><b>CONCLUSION</b>Infiltrating mast cells up-regulates p21 to promote NED and increase docetaxel resistance in PCa cells in vitro.</p>

Three-dimensional versus two-dimensional imaging systems in laparoscopic radical prostatectomy for prostate cancer: a retrospective cohort study / 南方医科大学学报

<p><b>OBJECTIVE</b>To compare the perioperative, functional and oncologic outcomes of patients with prostate cancer receiving laparoscopic radical prostatectomy (LRP) using three-dimensional (3D) versus two-dimensional (2D) imaging systems.</p><p><b>METHODS</b>From February, 2014 to January 2016, 72 consecutive patients with clinically localized prostate cancer underwent LRP with 2D or 3D imaging systems performed by a single experienced surgeon. The baseline characteristics, perioperative data, and functional and oncologic outcomes of the patients were collected and analyzed.</p><p><b>RESULTS</b>Thirty-six patients underwent 3D LRP and the other 36 patients underwent 2D LRP. Compared with 2D LRP group, 3D LRP group had a significantly shorter operative time (167 vs 218 min, P<0.001), a smaller volume of intraoperative blood loss (86.11 vs 177.78 mL, P<0.001) and a better early urinary continence outcome (88.89% vs 63.89%, P=0.026). No significant differences were found between the two groups in terms of complications, potency outcome or biochemical recurrence-free rate.</p><p><b>CONCLUSION</b>Compared with 2D LRP, 3D LRP shortens the operative time, reduces intraoperative blood loss and is associated with a better early urinary continence outcome in patients with clinically localized prostate cancer.</p>

Construction of a recombinant lentiviral vector of p38 MAPK and establishment of a human prostatic carcinoma cell line stably expressing p38 MAPK / 南方医科大学学报

<p><b>OBJECTIVE</b>To construct a recombinant lentiviral vector for p38 MAPK and establish a human prostatic carcinoma cell line that stably expresses p38 MAPK.</p><p><b>METHODS</b>EGFP/p38 fusion gene was subcloned into the lentiviral vector pTYF- EF1α-IRES-EGFP. The recombinant lentiviral vector pTYF-EF1α-EGFP/p38 was indentified by restriction enzyme digestion, and packaged in HEK 293T cells using lipofectamintm2000 with the packaging plasmid psPAX2 and envelope plasmid pMD2.G. The viral titer was tested according to the expression level of GFP. The resulting recombinant lentiviral vector was transduced into human prostatic carcinoma DU145 cells, and stably transduced cells were selected by limiting dilution analysis. The intracellular expression level of total p38 was detected by Western blotting and the cell growth curve was drawn.</p><p><b>RESULTS</b>DNA restriction enzyme digestion demonstrated that the recombinant lentiviral vector of the fusion gene EGFP/p38 (pTYF-EF1α-EGFP/p38) was constructed successfully. The recombinant lentiviral vector was packaged in 293T with a viral titer of 4.7×10(6) TU/ml. A stable cell line, EGFP/p38-DU145, was established, which stably expressed exogenous EGFP/p38 MAPK fusion protein as detected by Western blotting and showed a lowered growth rate compared to the control cells.</p><p><b>CONCLUSION</b>We have successfully constructed a recombinant lentiviral vector of the fusion gene EGFP/p38 and established a stable cell line EGFP/p38-DU145. Overexpression of p38 has a significant inhibitory effect on the proliferation of DU145 cells in vitro.</p>

A comparative analysis of the methods for titering adenoviruses / 南方医科大学学报

<p><b>OBJECTIVE</b>To compare different methods commonly used for titering adenovirus and analyze the advantages and limitations of each method.</p><p><b>METHODS</b>Four recombined adenoviruses (Ad-G-AT2R-EGFP, Ad-CMV-EGFP, Ad-mif-shRNA-EGFP and Ad-CBA-GFP) were amplified and purified, and each was titered by optical absorbance, real-time PCR, green fluorescent protein (GFP)-labeled method, immunoassay, and cytopathic effect (CPE). The results were then comparatively analyzed.</p><p><b>RESULTS</b>No significant difference was found in the titer amounts derived from GFP-labeled method, immunoassay, and cytopathic effect method (P>0.1). A positive correlation was noted in the titer amounts determined by real-time PCR and immunoassay (r=0.965), even though the value (vg/ml) obtained by real-time PCR was 10 times higher than that by immunoassay (ifu/ml).</p><p><b>CONCLUSION</b>GFP-labeled method and immunoassay allow rapid determination of the adenoviral titer. Real-time PCR can not directly determine the real infectious titer of the adenovirus, but the result is well correlated to that of immunoassay and reflects, though indirectly, the actual infectious titer of adenovirus. Considering the procedural convenience and shorter time consumption, real-time PCR is still a practical method for adenoviral titration.</p>

Rapid construction of a mammalian cell surface display library of full-length human antibodies / 南方医科大学学报

<p><b>OBJECTIVE</b>To construct a mammalian cell surface display library of full-length human antibodies.</p><p><b>METHODS</b>The total RNA was isolated from human peripheral blood mononuclear cells (PBMCs), and the genes encoding the heavy chain variable regions and kappa light chains (VH and Cκ) of the antibodies were amplified by RT-PCR. The amplified VH and Cκ gene sequences were separately inserted into the vector pDGB-HC-TM. The ligation mixtures were transformed into competent E.coli DH5α cells to construct the antibody libraries, and the library sizes and diversity were analyzed. The library DNAs were transfected into CHO cells and the expression of the full-length human antibodies on the surface of CHO cells was analyzed by flow cytometry.</p><p><b>RESULTS</b>The heavy chain gene library constructed showed a diversity of 2.6 × 10(5), and the kappa light chain gene library had a diversity of 2.0 × 10(5). Sequence analysis of 10 clones randomly selected from the constructed heavy chain gene library and 10 from the light chain gene library showed that 8 heavy chain clones and all 10 light chain clones contained correct open reading frames. Flow cytometry demonstrated that all the 18 clones expressed full-length antibodies, which could be detected on CHO cell surfaces. After co-transfection of the heavy chain and light chain gene libraries into CHO cells, the expression of full-length antibodies on CHO cell surfaces was detected with the positive cells reaching 31.5.</p><p><b>CONCLUSIONS</b>A full-length human mammalian display antibody library with a combinatory diversity of 5.2 × 10(10) can be constructed in two weeks, which allows the display of full-length antibodies on mammalian cell surface.</p>

Expression of Gluc-Fluc dual luciferase plasmid after transfection into MB49 bladder cells / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the expression characteristics of Gluc-Fluc dual luciferase plasmid after its transfection into MB49 bladder cells.</p><p><b>METHODS</b>pAAV2neoCAG-Gluc-2A-Fluc and pAAV2neo-Gluc plasmids were separately transfected into MB49 cells via LipofectamineTM2000. The Gluc activity in the cell culture supernatant and the Fluc activity in the cells were detected by luminometer and Lumina Imaging system.</p><p><b>RESULTS</b>The luminometer result showed that the activity of Gluc in the supernatant increased gradually in a cell number- and time-dependent manner, while Fluc activity in the cells increased with the cell number but not with time. The Lumina Imaging system showed that Gluc-Fluc was successfully expressed in MB49 bladder cells and cell lines with stable Gluc-Fluc expression were obtained after G418 selection.</p><p><b>CONCLUSION</b>Gluc in the dual luciferase plasmid retains its expression characteristics. Due to the advantages of Fluc in localization in living imaging and the easy quantitative detection of Gluc, the dual luciferase plasmid, after transfection in MB49 bladder cells, allows reliable and dynamic detection of tumor growth in animal models.</p>

Patient outcome and prognostic factors of renal cell carcinoma in clinical stage T(1-3)N(1-2)M(0): a single-institution analysis / 南方医科大学学报

<p><b>OBJECTIVE</b>To report our data of patients with clinical stage T(1-3)N(1-2)M(0) renal cell carcinoma (RCC) and explore the biological behavior of this malignancy.</p><p><b>METHODS</b>A total of 531 patients with no distant metastatic RCC underwent open radical nephrectomy at our institution between 1988 and 2008, among whom 42 patients with histological nodal metastases had successful surgical tumor resection. The clinical data and outcomes of the 42 patients were analyzed.</p><p><b>RESULTS</b>Of those 42 patients, 19.0% had T1, 21.4% had T2, and 59.5% had T3 stage tumors; 42.9% had N1 and 57.1% had N2 stage tumors. Tumor recurred in 30 (71.4%) patients after the surgery, and death occurred in 26 (61.9%) cases at the last follow-up; among the recurrent cases, 83.3% (25/30) had multiple metastases at the initial recurrence. The median cancer-specific survival (CSS) and disease-free survival (DFS) was 23 and 11 months in these cases, respectively. Multivariate analysis demonstrated that Fuhrman grade (P=0.005), N stage (P=0.014) and T stage (P=0.037) were the independent predictors of CSS; Eastern Cooperative Oncology Group (ECOG) performance status (PS) (P=0.002), tumor size (P=0.007), Fuhrman grade (P=0.009) and N stage (P=0.019) were the independent predictors of DFS.</p><p><b>CONCLUSION</b>Patients with T(1-3)N(1-2)M(0) RCC have poor prognosis. N stage is an independent predictor of both CSS and DFS, suggesting that extended lymph node dissection should be performed when suspicious enlarged nodal disease is found during surgery.</p>

Construction of a dual-luciferase co-expression vector and its characteristics in vitro and in vivo / 病毒学报

A novel dual luciferase expression vector was designed and its expression characteristics were studied in vitro and in vivo. Firstly, the Gluc and Fluc genes were connected via the TaV 2A sequence by overlaping PCR, and inserted into the expression vector pAAV2neoCAG, obtaining the recombinant plasmid pAAV2neoCAG-Gluc-2A-Fluc. Then pAAV2neoCAG-Gluc-2A-Fluc was transfected into BHK21 cells and the activity of Gluc and Fluc in the supernatant and cell lysates were assayed respectively. Results showed that both Gluc and Fluc were expressed successfully. The Gluc was mainly detected in the culture media while the Fluc was mostly within cells. The activity of Gluc in the supernatant increased gradually with time while the Fluc activity in cells almost kept stable. To investigate the expression of pAAV2neoCAG-Gluc-2A-Fluc in vivo, the plasmid was hydrodynamically injected into BALB/c mice through tail vein. The Gluc activity was assayed in a small volume of blood taken by tail vein at different time points. Results showed that Gluc was expressed stably at least 7 days. Live bioluminescence imaging technology was used to compare the expression characteristics of Gluc and Fluc. Whole body imaging was seen when coelenterazine, a specific substrate for Gluc, was injected, and the imaging signals decreased rapidly within 10 minutes. Liver imaging was showed when Flue specific substrate named D-Luciferin was injected, and the imaging remained stable at least for half an hour. The dual luciferase expression vector pAAV2neoCAG-Gluc-2A-Fluc combines the advantages of the secreted report gene Gluc and the non-secreted report gene Fluc, and will provide a new tool for cell labeling and tracing.

Immobilization of streptavidin-tagged bioactive hTNF-alpha on biotinylated mucosal surface of the bladder wall for treatment of superficial bladder cancer in mice / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate a novel immunotherapy through immobilization of streptavidin-tagged hTNF-alpha on the biotinylated mucosal surface of the bladder wall for bladder cancer treatment in mice.</p><p><b>METHODS</b>A total of 120 female C57BL/6j mice were randomized into 5 equal groups, namely blank control, PBS, soluble hTNF-alpha, SA-GFP, and SA-hTNF-alpha treatment groups. Twenty-four hours after establishment of a mouse model of orthotopic superficial bladder cancer, SA-hTNF-alpha fusion protein was immobilized on the biotinylated mucosal surface of the bladder wall, which was repeated every 4 days for a total of 6 sessions. Immunohistochemistry was performed to detect the retention time of SA-hTNF-alpha fusion protein in the biotinylated mouse bladder mucosa and the distribution of CD4(+) and CD8(+) lymphocytes in the mucosa and tumor tissues, with the tumor growth and mouse survival also observed. The cytotoxiciy of the tumor-specific lymphocytes was evaluated. The mice responding well to the treatment were re-challenged by MB49 and monitored for survival.</p><p><b>RESULTS</b>SA-hTNF-alpha could be efficiently and stably immobilized on the bladder mucosal surface for as long as 7 days. On day 60 after MB49 implantation, 18 out of 22 SA- hTNF-alpha-treated mice survived, with 9 appearing tumor-free, but all the mice in PBS control group died. Five out of 9 tumor-free mice in SA-hTNF-alpha group showed resistance to a re-challenge with intravesical MB49. The numbers of CD4(+) and CD8(+) lymphocytes were significantly greater in SA-hTNF-alpha group than in the other groups (P<0.05). The cytotoxicity of the tumor-specific lymphocytes was significantly stronger in SA-hTNF-alpha group than in the other groups (P<0.05).</p><p><b>CONCLUSION</b>SA-hTNF-alpha immobilized on the biotinylated mucosal surface of the bladder wall can significantly inhibit the tumor growth and promote the survival of the mice bearing orthotopic superficial bladder cancer.</p>

Construction of human full-length renal cell carcinoma patient-specific antibody library by mammalian cell surface display / 南方医科大学学报

<p><b>OBJECTIVE</b>To construct human renal cell carcinoma patient-specific full-length antibody library using mammalian cell surface display technique.</p><p><b>METHODS</b>Peripheral blood mononuclear cells (PBMC) were isolated from patients with renal cell carcinoma. The repertoires of kappa light chain (LCkappa) and heavy chain variable region (VH) of antibody were amplified by RT-PCR. The LCkappa and VH libraries were inserted into the vector pDGB-HC-TM separately, and the ligated libraries were transformed into competent E.coli TOPO10 to construct the renal cell carcinoma patient-specific antibody heavy and light chain libraries. 293T cells were co-transfected with the libraries and the full-length human antibodies expressed on the surface of 293T cells were analyzed by flow cytometry.</p><p><b>RESULTS</b>The libraries of renal cell carcinoma-specific antibody kappa light chain (LCkappa) and heavy chain (IgG1) were constructed. The expression of the full-length human antibodies on the surface of 293T cell was confirmed by flow cytometry. The libraries showed an expressible combinatory diversity of 7.5x10(10).</p><p><b>CONCLUSION</b>The expressible antibody library provides a useful platform for screening of renal cell carcinoma-specific antibodies.</p>

Predictive value of fluorescence in situ hybridization in patients with bladder cancer / 南方医科大学学报

<p><b>OBJECTIVE</b>To assess the value of fluorescence in situ hybridization (FISH) in the diagnosis of bladder cancer.</p><p><b>METHODS</b>Urine samples from 100 patients suspected of having bladder cancer were collected before cystoscopy for immediate urine cytology and FISH analysis. The criteria for FISH abnormality were determined by evaluating the urine specimens from 20 subjects without urogenital neoplasm.</p><p><b>RESULTS</b>The overall sensitivity of cytology and FISH was 43.2% and 82.4%, and their specificity was 92.3% and 88.5%, with diagnostic concordance rate of 56.0% and 84.0%, respectively. The differences between FISH and cytology showed statistical significance in the sensitivity, diagnostic concordance rate, non-muscle-invasive cancer and primary cancer.</p><p><b>CONCLUSION</b>The sensitivity and efficiency of FISH in the detection of bladder cancer are superior to those of cytology, especially for prophase cancer.</p>

Correlation of aging with psychological and organic ED: nocturnal electrobioimpedance volumetric assessment of 83 cases / 中华男科学杂志

<p><b>OBJECTIVE</b>The ratio of psychological to organic ED changes with aging. This study aimed to analyze the results of nocturnal electrobioimpedance volumetric assessment (NEVA) for ED patients of different age groups and their significance in the diagnosis of ED.</p><p><b>METHODS</b>A total of 83 ED patients were divided into 4 age groups (< or = 29 yr, 30 -39 yr, 40 -49 yr and > or = 50 yr) and detected for nocturnal penile tumescence (NPT) by NEVA.</p><p><b>RESULTS</b>Thirty-four of the cases were diagnosed as organic ED, and the other 49 as psychological ED. With the increase of age, the former was increased from 30.3% in the < or = 29 yr group to 60.0% in the > or = 50 yr group, while the latter decreased from 69.7% to 40.0%.</p><p><b>CONCLUSION</b>The percentage of organic ED tends to grow with the increase of age, while that of psychological ED is just the opposite.</p>

Efficacy of low-dose tadalafil on ED assessed by Self-Esteem and Relationship Questionnaire / 中华男科学杂志

<p><b>OBJECTIVE</b>To explore the effects of low-dose oral tadalafil on self-esteem, confidence and sexual relationship in ED patients.</p><p><b>METHODS</b>We treated 17 ED patients with oral tadalafil at the low dose of 5 mg once daily for 12 weeks, and used the paired t test to compare their scores on The Self-Esteem and Relationship Questionnaire (SEAR) and IIEF-5 and the results of nocturnal penile tumescence (NPT) obtained by nocturnal electrobioimpedance volumetric assessment (NEVA) before and after the medication.</p><p><b>RESULTS</b>The scores on SEAR and IIEF-5 were significantly increased (P < 0.01) and NPT markedly improved (P < 0.05) after tadalafil treatment as compared with the baseline.</p><p><b>CONCLUSION</b>Low-dose oral tadalafil once daily can significantly improve the self-esteem, confidence, sexual relationship satisfaction and NPT of ED patients.</p>

A simple and efficient method for establishing a mouse model of orthotopic MB49 bladder cancer / 南方医科大学学报

<p><b>OBJECTIVE</b>To establish a simple and efficient method for establishing a mouse model of orthotopic superficial bladder cancer.</p><p><b>METHODS</b>C57BL/6 mice were anesthetized with sodium pentobarbital and catheterized with modified IV catheter (24 G). The mice were intravesically pretreated with HCl and then with NaOH, and after washing the bladders with phosphate-buffered saline (PBS), 100 microl (1 x 10(7)) MB49 cells were infused and allowed to incubate in the bladder for 2 h followed intravesical mitomycin C (MMC) administration. The tumor formation rate, survival, gross hematuria, and bladder weight were determined as the outcome variables, and the pathology of the bladders was observed.</p><p><b>RESULTS</b>Instillation of MB49 tumor cells resulted in a tumor formation rates of 100% in all the pretreated groups while 0% in the control group without pretreatment. MMC significantly reduced the bladder weight as compared to PBS.</p><p><b>CONCLUSION</b>We have successfully established a stable, reproducible, and reliable orthotopic bladder cancer model in mice.</p>

Diagnosis and treatment of transverse testicular ectopia: a case report and literature review / 南方医科大学学报

<p><b>OBJECTIVE</b>To study the etiology, diagnosis and treatment of transverse testicular ectopia (TTE).</p><p><b>METHODS</b>A case of TTE was treated with orchidopexy.</p><p><b>RESULTS</b>Six months after the operation, both of the two testes were in proper positions with normal vascular supply.</p><p><b>CONCLUSION</b>TTE is a rare congenital abnormality of the male reproductive system with unknown etiology, and surgical correction remains the best option for treatment.</p>

Identification of human testicular embryonal carcinoma proteins by two-dimensional gel electrophoresis and mass spectrometry / 南方医科大学学报

<p><b>OBJECTIVE</b>To separate and identify human testicular embryonal carcinoma proteomics using two-dimensional electrophoresis (2-DE) and mass spectrometry.</p><p><b>METHODS</b>Immobilized pH gradient two-dimensional polyacrylamide gel electrophoresis was used to separate the total proteins of the samples. After silver staining, PDQuest 7.30 image analysis software was applied to analyze the 2-DE images. Three of the proteins highly expressed in human testicular embryonal carcinoma were identified by matrix-assisted laser adsorption/ionization-time of flight-tandem mass spectrometry (MALDI-TOF-MS/MS).</p><p><b>RESULTS</b>2-DE effectively screened the differentially expressed proteins in the carcinoma tissues. Three proteins highly expressed in the carcinoma were successfully identified.</p><p><b>CONCLUSION</b>The proteins of human testicular embryonal carcinoma can be effectively separated and analyzed using 2-DE and mass spectrometry. Proteomic analysis offers a new means for further study of this carcinoma.</p>

Reduced expression of programmed cell death 5 protein in tissue of human prostate cancer / 中国医学科学杂志(英文版)

<p><b>OBJECTIVE</b>To investigate the expression of programmed cell death 5 (PDCD5) in tissues of normal human prostate (NP), benign prostatic hyperplasia (BPH), and prostate cancer (PCa) in order to assess the clinical role of PDCD5 in PCa.</p><p><b>METHODS</b>PDCD5 expression was determined by EnVision immunohistochemical staining in formalin-fixed and paraffin-embedded specimens obtained from 12 subjects with NP, 22 with BPH, and 22 with PCa. In addition, PCa cases were classified as low/middle-risk (Gleason sum < or = 7) and high-risk (Gleason sum >7) on the basis of Gleason grade. Positive expression rates and intensity of PDCD5 protein were observed under light microscope and analyzed with computer imaging technique. Expression of PDCD5 was compared among different prostatic tissues.</p><p><b>RESULTS</b>The expression of PDCD5 was significantly lower in tissue of PCa than in tissues of NP and BPH (P<0.01). However, there was no significant difference in PDCD5 expression between tissues of NP and BPH. In addition, the expression of PDCD5 was further downregulated with the increase of Gleason sum in PCa.</p><p><b>CONCLUSIONS</b>By downregulating apoptosis, low PDCD5 expression may play an important role in the occurrence and development of PCa. PDCD5 is supposed to have a potential clinical value to be a new predictor of progression and target of gene therapy in PCa.</p>

Expression of human telomerase reverse transcriptase in renal cell carcinoma and its clinical significance / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the expression of human telomerase reverse transcriptase (hTERT) in renal cell carcinoma (RCC) and its clinical significance.</p><p><b>METHODS</b>The expression levels of hTERT mRNA and protein were detected using RT-RCR and Western blotting in 45 RCC tissues, 45 adjacent tissues and 786-0 cell line, and the associations of hTERT expression with the tumor size, clinical stage, pathological type and grade were evaluated.</p><p><b>RESULTS</b>hTERT mRNA and protein was expressed at significantly higher levels in RCC tissues than in the adjacent tissues (P=0.000), and no correlation of hTERT expression was found with the tumor size, clinical stage, pathological type or grade.</p><p><b>CONCLUSION</b>hTERT might serve as a diagnostic and prognostic marker for RCC, and also shed light on the new clues for gene therapy of RCC.</p>

Effect of adenovirus-mediated TK/GCV gene therapy in combination with TNF-alpha against murine bladder cancer cells in vitro / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the cell-killing effect of adenovirus-mediated TK-ganciclovir (GCV) gene therapy in combination with tumor necrosis factor-alpha (TNF-alpha) against murine bladder carcinoma cells in vitro.</p><p><b>METHODS</b>Murine bladder carcinoma MB49 cells were transfected with the adenoviral vector containing TK gene and green fluorescent protein (GFP) gene. The transfection efficiency was observed and the TK gene expression in the transfected cells was detected by RT-PCR. The survival rate of MB49 cells in response to TNF-alpha treatment and that of the TK gene-transfected cells after treatment with GCV and GCV+TNF-alpha were determined by MTT assay. The apoptosis of the cells after the treatments was analyzed by flow cytometry.</p><p><b>RESULTS</b>In cells transfected with TK gene, the cell inhibition rate increased gradually with the increment of GCV and TNF-alpha concentration. GCV in combination with TNF-alpha resulted in significantly increased killing efficiency of the cells as compared with GCV or TNF-alpha treatment alone, and the effect of the combined treatment was enhanced as the TNF-alpha concentration increased. GCV treatment (50 microg/ml) alone produced a cell killing rate of (24.39-/+1.10)%, and when combined with 5 microg/ml TNF-alpha, the rate was increased to (40.05-/+0.97) %, and further to (65.47-/+0.67) % when TNF-alpha concentration increased to 20 microg/ml. Flow cytometry revealed obvious apoptosis of the cells 8 h after treatments with TK/GCV, TNF-alpha, or TK/GCV+TNF-alpha, and the combined treatment resulted in the highest cell apoptotic rate.</p><p><b>CONCLUSION</b>TK/GCV in combination with TNF-alpha can enhance the effect of suicide gene therapy against murine bladder carcinoma cells and effectively induce apoptosis of the cells.</p>