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1.
National Journal of Andrology ; (12): 207-212, 2014.
Article in Chinese | WPRIM | ID: wpr-309734

ABSTRACT

<p><b>OBJECTIVE</b>To construct a recombinant adenovirus expression vector containing the anti-oncogene PTEN and to investigate the effects of the PTEN gene on the proliferation of prostate cancer PC-3 cells and the expressions of cyclin D1 and p21 in the PC-3 cells.</p><p><b>METHODS</b>The PTEN gene was amplified from the rat hippocampus by RT-PCR and cloned into the shuttle plasmid pEN-TR2A. The plasmids were constructed and amplified in 293A cells. Prostate cancer PC-3 cells were cultured in vitro and infected with the adenoviral vector carrying the PTEN gene (Ad-PTEN). The up-regulation of the PTEN protein was measured by indirect immuno-fluorescence assay; the expressions of PTEN, cyclin D1 and p21 in the cells infected with Ad-PTEN and Ad-LacZ were determined by</p><p><b>RESULTS</b>The Western blot; and the effect of PTEN on the cell proliferation was detected by MTT assay and plate colony formation. recombinant adenoviral vector Ad-PTEN was successfully constructed. Western blot showed a significantly increased expression of the PTEN protein in the PC-3 cells infected with Ad-PTIEN (0.215 +/-0.065) as compared with that in the control ([0.052 +/-0.009], t = 4. 30, P <0.05) and the Ad-LacZ group ( [0. 056 +/- 0.008 ] , t =4.21, P <0.05). The expression of cyclin D1 was significantly lower in the Ad-PTEN-infected PC-3 cells (0. 256 +/- 0. 072) than in the control ( [0. 502 +/- 0. 087 ], t = 3.77, P < 0.05) and the Ad-LacZ group ([0.498 +/-0.081] , t =3.87, P <0.05), while the expression of p21 remarkably higher in the Ad-PTEN-infected PC-3 cells (0.589 +/-0. 076) than in the control ([0. 146 +/-0.026] , t = 9.55, P<0. 01) and the Ad-LacZ group ([0. 163 +/-0. 024] , t = 9.26, P <0.01). Ad-PTEN significantly inhibited the growth of the PC-3 cells (21.98%) at 48 h (t = 6.80, P <0.01). The colony formation rate of the PC-3 cells was (37.4 +/-4. 18)% in the Ad-PTEN group, significantly lower than (54.9 +/-4.81)% in the control (t =4.76, P<0.01) and (56.5 +/- 5.42)% in the Ad-LacZ group (t=4.83, P<0.01).</p><p><b>CONCLUSION</b>The expression of PTEN induced by Ad-PTEN can significantly inhibit the proliferation of PC-3 cells, down-regulate the expression of cyclin D1, and up-regulate the expression of p21.</p>


Subject(s)
Animals , Humans , Male , Rats , Adenoviridae , Genetics , Cell Line, Tumor , Cell Proliferation , Cyclin D1 , Metabolism , Cyclin-Dependent Kinase Inhibitor p21 , Metabolism , PTEN Phosphohydrolase , Genetics , Prostatic Neoplasms , Metabolism , Pathology , Rats, Sprague-Dawley
2.
National Journal of Andrology ; (12): 884-887, 2011.
Article in Chinese | WPRIM | ID: wpr-305771

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the effects of staurosporine (ST) on the proliferation and apoptosis of prostate cancer PC-3 cells.</p><p><b>METHODS</b>Prostate cancer PC-3 cells were treated in vitro with ST at 10(-8) mol/L. The expressions of cyclin A and cyclin D1 proteins in the cells were detected by Western blot, the effect of ST on the proliferation of the cells determined by MTT assay and plate colony formation, the apoptosis of the cells examined by flow cytometry, and their morphological changes observed under the light microscope.</p><p><b>RESULTS</b>ST treatment markedly decreased the expressions of cyclin A and cyclin D1 in the PC-3 cells, and significantly inhibited the growth of the PC-3 cells (19.35%) at 48 h. (F = 31.06, P < 0.01). The colony formation rate of the PC-3 cells was (37.10 +/- 3.43) % in the ST group, significantly lower than (64.80 +/- 4.34) % in the control (chi2 = 14.59, P < 0.05) and (62.80 +/- 4.36) % in the DMSO group (chi2 = 12.50, P < 0.05), while the apoptosis rate of the cells was remarkably higher in the ST group ([19.6 +/- 2.20] %) than in the control ([5.33 +/- 1.40] %) and the DMSO group ([5.50 +/- 0.96] %) (F = 104.36, P < 0.01). Under the light microscope, the ST-treated cells were round with indistinct margins as compared with those of the other two groups.</p><p><b>CONCLUSION</b>ST could significantly inhibit the proliferation and induce the apoptosis of PC-3 cells.</p>


Subject(s)
Humans , Male , Apoptosis , Cell Line, Tumor , Cell Proliferation , Prostatic Neoplasms , Pathology , Staurosporine , Pharmacology
3.
National Journal of Andrology ; (12): 905-910, 2010.
Article in Chinese | WPRIM | ID: wpr-266247

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the effects of intraprostatic injection of botulinum toxin A (BTX-A) on benign prostate hyperplasia (BPH) in rats.</p><p><b>METHODS</b>Models of BPH were established in adult male Sprague-Dawley rats by injection of testosterone propionate, and then divided into three BTX-A groups, injected with BTX-A into the ventral prostate at the doses of 5 U, 10 U and 20 U, a negative control group, injected with saline only, and a sham operation group, with 12 in each. The prostates of the animals were harvested at 2 or 4 weeks after the injection, their volumes and weights measured, histological changes examined by HE staining, and glandular and interstitial areas semi-quantified by the image analysis system.</p><p><b>RESULTS</b>Two rats died in the 20 U group within 3 days after BTX-A injection. Compared with the saline group, the 5 U, 10 U and 20 U BTX-A groups showed significant decreases in prostatic volume (P < 0.01, 0.01 and 0.05), weight, and glandular and interstitial areas as well as atrophic epithelia in the glandular tube at 2 weeks. These changes were lessened at 4 weeks, especially in the 5 U group.</p><p><b>CONCLUSION</b>Intraprostatic injection of BTX-A induces obvious atrophy and histological changes of the prostate, but meanwhile may potentially result in death at a large dose.</p>


Subject(s)
Animals , Male , Rats , Botulinum Toxins, Type A , Therapeutic Uses , Toxicity , Prostate , Pathology , Prostatic Hyperplasia , Drug Therapy , Pathology , Rats, Sprague-Dawley
4.
National Journal of Andrology ; (12): 224-226, 2008.
Article in Chinese | WPRIM | ID: wpr-319241

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the expressions of the aging gene P16(ink4a) and anti-aging gene HST2 in benign prostatic hyperplasia (BPH).</p><p><b>METHODS</b>Twenty-three BPH and eighteen normal prostate specimens were collected and total RNA was extracted, followed by the reverse transcriptase polymerase chain reaction (RT-PCR). The expressions of P16(ink4a) was detected by semi-quantitative analysis in BPH and normal prostate tissues.</p><p><b>RESULTS</b>P16(ink4a) mRNA, rather than HST2, was expressed in the BPH and normal prostate tissues. Semi-quantitative analysis showed that the P16(ink4a) mRNA expression in the normal prostate tissues (0.4868 +/- 0.545 was significantly higher than in the BPH tissues (0.2783 +/- 0.0268, with a statistical difference in between (P < 0. 05).</p><p><b>CONCLUSION</b>P16(ink4a) might play an important role in the pathogenesis of BPH and is probably one of the factors of cell aging escape.</p>


Subject(s)
Adult , Aged , Humans , Male , Cyclin-Dependent Kinase Inhibitor p16 , Genetics , Fibroblast Growth Factor 6 , Genetics , Gene Expression Profiling , Pilot Projects , Prostatic Hyperplasia , Genetics , Pathology , RNA, Messenger , Genetics , Metabolism , Reverse Transcriptase Polymerase Chain Reaction
5.
National Journal of Andrology ; (12): 664-669, 2005.
Article in Chinese | WPRIM | ID: wpr-339455

ABSTRACT

<p><b>OBJECTIVE</b>To evaluate the effect of the levels of IGF-I in the epididymis and the expression of IGF-I in the testis of adult male rat after the administration of cyclophosphamide.</p><p><b>METHODS</b>Ninety-six male adult rats (8 weeks age) were divided into 6 groups. The doses given to the rats of the groups 1 to 5 were 10, 20, 40, 80 and 100 mg/(kg x d), respectively. The remaining group was served as control. All those rats were sacrificed and IGF-I were quantitatively determined by ELISA techniques 2 and 4 weeks after the administration of the drug (by gastric fudge). Immunohistochemical SP technique was used to examine expression of IGF-I in rat testis.</p><p><b>RESULTS</b>The levels of cell factors (IGF-I) in the epididymis of the rats were gradually reduced with the increasing time and dose after administration of the drug. In the mean time the expression of IGF-I in the tissues of the testis of those rats were also gradually reduced.</p><p><b>CONCLUSION</b>In the time of oligozoospermia/azoospermia induced by the administration of cyclophosphamide, the expression levels of IGF-I in the genetic system were significantly reduced. The possible mechanism of these changes could be attributed to the lower spermatogenesis function of the testis caused by the administration of cyclophosphamide.</p>


Subject(s)
Animals , Male , Rats , Azoospermia , Metabolism , Cyclophosphamide , Toxicity , Disease Models, Animal , Enzyme-Linked Immunosorbent Assay , Epididymis , Metabolism , Immunohistochemistry , Insulin-Like Growth Factor I , Oligospermia , Metabolism , Rats, Sprague-Dawley , Testis , Metabolism
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