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1.
National Journal of Andrology ; (12): 679-683, 2010.
Article in Chinese | WPRIM | ID: wpr-295019

ABSTRACT

<p><b>OBJECTIVE</b>To construct an oncolytic adenovirus with the DD3 promoter regulation, expressing small hairpin RNA and targeting the SATB1 gene (SATBI-shRNA), and to evaluate its potential for inhibiting the growth of human prostatic carcinoma cells (LNCaP) in vitro.</p><p><b>METHODS</b>SATB1-shRNA expression cassettes containing the H1 promoter were produced by PCR from pSilencer3. 1-SATB1 and inserted into the pZD55 vector, and the recombinant plasmid pZD55-SATB1-shRNA was constructed, pZD55SATB1-shRNA and pZXC2-DD3-E1A were digested with EcoRV and Xba I , and the obtained expression cassettes linked each other to construct the recombinant plasmid pDD3-ZD55-SATB1, which was cotransfected with the pBHGE3 packaging plasmids mixture into 293 cells by Effectence. The recombined adenoviruses DD3-ZD55-SATB1 were identified by PCR. Viruses were propagated on HEK293 cells and purified by standard techniques, and the functional PFU titers determined by plaque assay on 293 cells. The antitumor potential of DD3-ZD55-SATB1 to LNCaP was evaluated by the crystal violet dye method. The expression level of the E1A gene was detected by Western blot, and that of the SATB1 gene in the adenovirus-infected LNCaP cells by both Western blot and RT-PCR.</p><p><b>RESULTS</b>An oncolytic adenovirus expressing SATB1-shRNA with the DD3 promoter regulation, DD3-ZD55-SATB1, was constructed successfully, whose functional PFU titer was 1.2 x 10(10) PFU/ml. DD3-ZD55-SATB1 showed an obvious cytopathic effect and a selective expression of E1A in the adenovirus-infected LNCaP cells; it exhibited a high LNCaP-targetability and specific SATB1-silencing effect.</p><p><b>CONCLUSION</b>The successful construction of the oncolytic adenovirus DD3-ZD55-SATB1 offers a new tool for researches on the gene therapy for human prostate cancer.</p>


Subject(s)
Humans , Male , Adenoviridae , Genetics , Carcinoma , Therapeutics , Cell Line, Tumor , Genetic Vectors , Matrix Attachment Region Binding Proteins , Genetics , Oncolytic Virotherapy , Methods , Oncolytic Viruses , Genetics , Promoter Regions, Genetic , Prostatic Neoplasms , Therapeutics , RNA Interference , RNA, Small Interfering , Genetics
2.
National Journal of Andrology ; (12): 1120-1123, 2009.
Article in Chinese | WPRIM | ID: wpr-252853

ABSTRACT

<p><b>OBJECTIVE</b>To improve the diagnosis and treatment of far advanced prostate cancer without clinically detectable bone metastasis.</p><p><b>METHODS</b>Cancer metastatic lesions were found in the liver and lungs respectively of two patients on routine medical examination, and only an abnormally elevated level of the serum prostate specific antigen (PSA) was observed in the following system examinations. The patients were diagnosed as having prostate cancer by prostate biopsy. MRI showed a discontinued prostate capsule, and ECT revealed no bone metastasis. Diagnostic treatment was conducted by giving LHRHa combined with antiandrogens. One of the patients underwent surgical castration at 12 months, and both received intensity modulated radiation therapy (80 Gy) at 15 and 18 months, respectively.</p><p><b>RESULTS</b>The metastatic lesions in the liver and lungs of the patients were either absent or significantly reduced after treated by maximal androgen blockade for 3 months, and all disappeared after 6 months'treatment, with the PSA level stabilized at less than 0.02 microg/L in one patient, and around 0.5 microg/L in the other. Antiandrogen treatment was suspended after radiotherapy. The results of liver, lung and bone scanning were normal during the 12-month follow-up, and the PSA level was below 1.0 microg/L.</p><p><b>CONCLUSION</b>Remote metastasis of prostate cancer may occur in ectosteal organs first, which deserves special attention. A combination of different treatment methods promises satisfactory results.</p>


Subject(s)
Aged , Humans , Male , Liver Neoplasms , Lung Neoplasms , Neoplasm Metastasis , Prostatic Neoplasms , Diagnosis , Pathology , Therapeutics
3.
Chinese Journal of Industrial Hygiene and Occupational Diseases ; (12): 90-92, 2003.
Article in Chinese | WPRIM | ID: wpr-265049

ABSTRACT

<p><b>OBJECTIVE</b>To observe the changes in peripheral blood T lymphocyte rDNA transcription activity and to study the significance of immune monitoring for patients with chronic benzene poisoning.</p><p><b>METHODS</b>Venous blood samples were withdrawn from 39 patients with chronic benzene poisoning, 20 patients with malignant disease and 22 healthy controls. Cell culture, argyrophil staining method, I-CLQ cell image analysis system were used in this study. rDNA transcription activity which was expressed by the ratio of integrated area (IA) of nucleolus to that of nucleus, and the ratio of integrated optical density (IOD) of argyrophilic nucleolus to that of argyrophilic nucleus.</p><p><b>RESULTS</b>(1) The value of IA and IOD in chronic benzene poisoning patients (7.95% +/- 1.13% and 7.15% +/- 1.15% respectively) were lower than those in controls (9.59% +/- 1.26% and 8.92% +/- 1.18% respectively), P < 0.01. The value of IA and IOD in chronic moderate benzene poisoning patients (6.54% +/- 0.88%) and (5.47% +/- 0.80%) were lower than those to be observed (7.98% +/- 1.06% and 7.13% +/- 0.96% respectively) as well as in mild poisoning patients (8.19% +/- 1.06% and 7.44% +/- 1.06% respectively), P < 0.05. (2) The value of IA and IOD in malignant group (4.10% +/- 1.50%, 3.67% +/- 1.42%) were much more lower than those in controls and in chronic benzene poisoning patients (P < 0.01).</p><p><b>CONCLUSION</b>rDNA transcription activity may be an index to monitor the cellular immune function of chronic benzene poisoning patients.</p>


Subject(s)
Adult , Female , Humans , Male , Middle Aged , Benzene , Poisoning , Chronic Disease , DNA, Ribosomal , Genetics , T-Lymphocytes , Metabolism , Transcription, Genetic
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