RESUMO
OBJECTIVE: To study the effects of MTA1 small interfering RNA (siRNA) on the anchorage-independent growth and anoikis of prostate cancer cell line PC-3. METHODS: After transfection of human prostate cancer PC-3 cells by MTA1 siRNA, we detected the expression of the MTA1 gene by real-time PCR and Western blot, the anchorage-independent growth of the cells by clone formation in soft agar, and their anoikis by DNA fragmentation assay and flow cytometry. RESULTS: Compared with the control group, MTA1 siRNA transfection significantly decreased the mRNA and protein levels of MTA1, inhibited the anchorage-independent growth of the PC-3 cells, and induced their anoikis, all in a dose- and time-dependent manner (r = 0.935, P = 0.001; r = 0.901, P = 0.0005; r = 0.916, P = 0.0003). CONCLUSION: MTA1 siRNA can inhibit the anchorage-independent growth of prostate cancer cells by inducing their anoikis.
Assuntos
Anoikis/genética , Histona Desacetilases/genética , Neoplasias da Próstata/genética , Proteínas Repressoras/genética , Regulação Neoplásica da Expressão Gênica , Humanos , Masculino , RNA Interferente Pequeno , Transativadores , Transfecção , Células Tumorais CultivadasRESUMO
OBJECTIVE: To explore the expression of pituitary tumor transforming gene (PTTG) in human renal clear cell carcinoma (RCCC) and its significance. METHODS: PTTG protein expression was detected by immunohistochemistry in 87 RCCC and 45 paired normal renal tissues. RESULTS: PTTG was expressed differentially between the normal renal and RCCC tissues. Compared with normal tissues, the primary RCCC tissues showed significantly increased expression of PTTG protein (P<0.001). The positive rate of PTTG protein was positively correlated to the tumor size, clinical stage and Fuhrman grade of RCCC (P=0.009, 0.008 and 0.035, respectively). CONCLUSION: Increased PTTG protein expression may be involved in the carcinogenesis and progression of RCCC.
Assuntos
Carcinoma de Células Renais/metabolismo , Neoplasias Renais/metabolismo , Proteínas de Neoplasias/metabolismo , Adulto , Idoso , Carcinoma de Células Renais/patologia , Feminino , Regulação Neoplásica da Expressão Gênica , Humanos , Imuno-Histoquímica , Neoplasias Renais/patologia , Masculino , Pessoa de Meia-Idade , Proteínas de Neoplasias/genética , SecurinaRESUMO
OBJECTIVE: To study the effect of laminarin sulphate (LAMS) on the expressions of PTEN and P27kip1 in androgen-independent prostate cancer PC-3 cells in vitro, and investigate the mechanism of its anti-tumor action. METHODS: The inhibitory effects of different concentrations of LAMS (0, 50, 100, 200 microg/ml) on androgen-independent prostate cancer PC-3 cells were detected by WST-8 assay. The morphology of PC-3 cells was observed under the fluorescence microscope, and the cell cycle and apoptosis were analyzed by flow cytometry. The mRNA and protein levels of PTEN and P27kip1 were measured by RT-PCR and Western blot. RESULTS: LAMS inhibited the proliferation of androgen-independent prostate cancer PC-3 cells in a dose- and time-dependent manner, and the cell cycle analysis showed that PC-3 cells were arrested in the S phase after treated with different concentrations of LAMS. The rate of apoptosis was increased and many typical apoptotic morphological features were observed under the fluorescence microscope. The PTEN and P27kip1 expressions at mRNA and protein levels were increased in a dose-dependent manner. CONCLUSION: LAMS can inhibit the proliferation, arrest the cell cycle in the S phase and induce apoptosis of prostate cancer PC-3 cells. The significantly increased expressions of PTEN and P27kip1 may be one of the mechanisms for LAMS inhibiting prostate cancer PC-3 cells.
Assuntos
Inibidor de Quinase Dependente de Ciclina p27/genética , PTEN Fosfo-Hidrolase/genética , Polissacarídeos/farmacologia , Neoplasias da Próstata/genética , Apoptose/efeitos dos fármacos , Ciclo Celular/efeitos dos fármacos , Linhagem Celular Tumoral/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Humanos , Masculino , Neoplasias da Próstata/sangue , RNA Mensageiro/genéticaRESUMO
OBJECTIVE: To investigate the effects of adenovirus-mediated PTEN and P27 on the invasion of PC-3 in vitro and angiogenesis, along with their synergy in the treatment of prostate cancer. METHODS: Recombinant adenovirus vectors of the human tumor suppressor genes PTEN and P27 were constructed. The replication-incompetent recombinant adenovirus was packaged and propagated in HEK293 cells. The viral titer was examined by plaque assay and the mRNA and protein expressions of PTEN and P27 in human prostate cancer cell line PC-3 infected with Ad-PTEN and Ad-P27 were determined by RT-PCR and Western blot respectively. The invasion of PC-3 cells in vitro was examined by Boyden chamber assay. MTT assay was used to testify the effect of supernatant from PC-3 infected with Ad-PTEN and Ad-P27 on the proliferation of endothelial cells ECV-304 and the CAM test was used to testify the effect of PTEN and P27 on angiogenesis. The difference between the combined therapy group and the single gene therapy group was also examined. RESULTS: The viral titers of Ad-PTEN and Ad-P27 were 1.8 x 10(7) pfu/ml and 1.2 x 10(9) pfu/ml respectively. Adenovirus infection verified that the mRNA and protein expression of PTEN and P27 were steady in human PC-3 cells. The invasion in vitro of PC-3 cells was significantly inhibited by infection with Ad-PTEN or/and Ad-P27. CAM and MTT assays of ECV-304 confirmed that the supernatant from PC-3 cells infected with Ad-PTEN or/and Ad-P27 could inhibit the angiogenesis effectively. There was a significant difference between the combined therapy group and the single gene therapy group. CONCLUSION: The combined gene therapy of Ad-PTEN and Ad-P27 plays a synergistic role in inhibiting the invasiveness of PC-3 cells and angiogenesis.
