RESUMO
OBJECTIVE: To evaluate the application of CXC chemokine receptor-4 (CXCR4) combined with alpha-methylacyl-CoA racemase (P504S) or P63 protein in the differential diagnosis of benign and malignant prostatic diseases. METHODS: The EnVision immunohistochemical method was used to detect the expressions of CXCR4, P504S and P63 protein in 40 specimens of PCa not treated by any anticancer therapy and 30 specimens of BPH tissues. The correlation was analyzed between CXCR4 expression and the characteristics of PCa metastasis. RESULTS: Of the 40 cases of PCa, 33 (82.5%) were stained positive for CXCR4, 37 (92.5%) for P504S and 2 (5%) for P63 protein. Of the 30 cases of BPH, 5 (16.6%) exhibited positivity for CXCR4, 1 for P504S and all for P63. P504S + P63 showed a higher rate of correct diagnosis of PCa than either CXCR4 + P63 or P504S + CXCR4. There was a statistically significant correlation between CXCR4 expression and cancer metastasis (P < 0.05). CONCLUSION: P504S, CXCR4 and P63 are useful tumor markers for the diagnosis and differentiation of benign and malignant prostatic diseases. CXCR4 gives a high rate of correct diagnosis when combined with P504S or P63, and has an important application value in the differential diagnosis of benign and malignant prostatic diseases.
Assuntos
Biomarcadores Tumorais/biossíntese , Proteínas de Membrana/biossíntese , Hiperplasia Prostática/diagnóstico , Neoplasias da Próstata/diagnóstico , Racemases e Epimerases/biossíntese , Receptores CXCR4/biossíntese , Idoso , Idoso de 80 Anos ou mais , Diagnóstico Diferencial , Humanos , Masculino , Pessoa de Meia-Idade , Hiperplasia Prostática/metabolismo , Hiperplasia Prostática/patologia , Neoplasias da Próstata/metabolismo , Neoplasias da Próstata/patologiaRESUMO
OBJECTIVE: To construct RNA interfering retrovirus vector targeting CXCR4 gene driven by human prostate-specific antigen promoter and investigate its targeted inhibition effects in androgen-responsive prostate cancer cells LNCaP. METHODS: To clone the CXCR4 targeting siRNA gene by PCR. The PCR products were inserted into the pGensil-1 plasmid containing U6 promoter and EGFP. U6 promoter was replaced by hPSA promoter. Then, the recombinant EGFP-hPSA-siCXCR4 fragment was sub-cloned into pLXSN, which was evaluated by restriction enzyme. The pLXSN-EGFP-hPSA-siCXCR4 was transfected into PA317 cells with Lipofectamine 2000. The virus obtained from transfected PA317 cells was transfected into PC-3m, LNCaP and MCF-7 cells, respectively. The expression of CXCR4 mRNA and protein was detected by RT-PCR and Western blot. The invasion ability of prostate carcinoma cells was detected by Transwell experiment. RESULTS: The recombinant pLXSN-hPSA-siCXCR4 was successfully constructed. The expression of CXCR4 mRNA and protein in LNCaP cells was blocked by pLXSN-hPSA-siCXCR4. The expression inhibition rate was (81.53 +/- 10.22)% at mRNA level detected by semi-quantitive RT-PCR and (90.52 +/- 9.31)% at protein level detected by Western blot, respectively, in LNCaP cells at 48 h. The expression of CXCR4 mRNA and protein was effectively inhibited by sequence-specific hPSA-siCXCR4 in LNCaP cells, but not in PC-3m and MCF-7 cells. The results of Transwell experiment showed that the number of cells in down-pore of micro-membrane was 139.9 +/- 14. 2 in the treated group, significantly less in comparison with 348.4 +/- 36. 4 in the controlled group (P < 0.05). However, the number of PC-3m and MCF-7 cells in down-pore of micro-membrane was not significantly different among the control and treated groups (P > 0.05). CONCLUSION: The downstream siRNA controlled by hPSA promoter in retrovirus system can be expressed selectively in androgen-responsive prostate carcinoma cells, showing an apparent targeting character. RNAi targeted to CXCR4 driven by hPSA promoter has a potential value in gene therapy of androgen-responsive prostate cancer.
Assuntos
Antígeno Prostático Específico/genética , Neoplasias da Próstata , Interferência de RNA , RNA Interferente Pequeno/genética , Receptores CXCR4/metabolismo , Animais , Linhagem Celular Tumoral , Regulação Neoplásica da Expressão Gênica , Vetores Genéticos , Humanos , Masculino , Camundongos , Células NIH 3T3 , Invasividade Neoplásica , Plasmídeos , Regiões Promotoras Genéticas , Neoplasias da Próstata/genética , Neoplasias da Próstata/metabolismo , Neoplasias da Próstata/patologia , RNA Mensageiro/metabolismo , Receptores CXCR4/genética , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo , Retroviridae/genética , TransfecçãoRESUMO
The safety and effectiveness of a novel Chinese one-shot dilation technique based on stimulated diuresis for percutaneous nephrolithotomy (PCNL) were investigated. After the feasibility of the Chinese one-shot dilation based on stimulated diuresis was verified by an animal study, this technique was applied in the clinical practice. A total of 67 patients in our department underwent the modified PCNL from July 2014 to June 2015. After the renal infundibulum was distended by stimulated diuresis, the kidney was punctured under the ultrasonographic guidance via the fornix of the target calyx. The working channel was dilated using a special designed pencil-shaped fascial dilator. The successful access rate, nephrostomy tract creation time, pre- and postoperative hemoglobin values and serum creatinine concentrations, stone-free rate and complications were recorded and analyzed. The renal infundibulum was successfully distended in all of the patients by the diuresis treatment. Under the ultrasonographic guidance, the successful access rate was 100% and the mean tract creation time was 2.0 min (range: 1.5-5.0 min). The stone-free rate right after surgery was 91.0%. Although the postoperative hemoglobin was significantly reduced (P<0.01), transfusion was not clinically necessary. There was no significant difference in serum creatinine concentrations before and after operation (P>0.05). No severe complication occurred during or after the PCNL. It was suggested that this Chinese one-shot dilation technique based on stimulated diuresis is an efficient and safe innovation for PCNL, and is even helpful for those patients with non-dilated pelvicaliceal systems.
Assuntos
Diurese , Nefrostomia Percutânea/métodos , Cirurgia Assistida por Computador/métodos , Adulto , Idoso , Animais , Creatinina/sangue , Feminino , Hemoglobinas/metabolismo , Humanos , Rim/cirurgia , Masculino , Pessoa de Meia-Idade , Nefrostomia Percutânea/efeitos adversos , Complicações Pós-Operatórias , Cirurgia Assistida por Computador/efeitos adversos , Suínos , UltrassonografiaRESUMO
OBJECTIVES: To examine the effects of suramin on the growth, cell cycle and apoptosis of a hormone refractory prostate cancer cell line PC-3M, and to explore the possible mechanisms. METHODS: The roles of diverse concentrations (10, 50, 100 and 200 mumol/L) of suramin on PC-3M cell proliferation at different ratios of fetal calf serum (FCS) (2%, 5%, 10%) were assayed respectively by trypan blue exclusion and tetrazolium (MTT) assay. The effect of suramin on cell cycle distribution and apoptosis induction of PC-3M cells was evaluated with flow cytometry (FCM). RESULTS: A higher dosage of suramin (200 mumol/L) had a cytotoxic effect on PC-3M cells, while lower dosages from 10 to 100 mumol/L produced a predominant inhibiting effect. Suramin could also play a growth suppressive role in the culture media containing 10% FCS, but to a much less extent than in the media containing lower concentrations(5%, 2%) of FCS. FCM analysis exhibited that suramin at a high dosage of 200 mumol/L could induce apoptosis, and at the other concentrations, G0/G1 cell cycle arrest. CONCLUSION: Suramin's proliferative suppression on PC-3M cells might result from several mechanisms including antagonistic action on growth stimulation via growth factor, arrest of cell cycle and induction of apoptosis.
Assuntos
Antineoplásicos/farmacologia , Proliferação de Células/efeitos dos fármacos , Neoplasias da Próstata/patologia , Suramina/farmacologia , Animais , Apoptose/efeitos dos fármacos , Bovinos , Ciclo Celular/efeitos dos fármacos , Linhagem Celular Tumoral , Relação Dose-Resposta a Droga , Sangue Fetal , Humanos , MasculinoRESUMO
PURPOSE: CXC chemokine receptor-4 (CXCR4) is closely involved in bone metastasis of prostate cancer, and CXCR4 levels are frequently increased in prostate cancer cells and tissues. In the present study, its biological effects on prostate cancer in vitro and in vivo and feasibility to be a therapy target were investigated using a RNA interfering retrovirus vector targeting CXCR4 gene driven by human prostate-specific antigen promoter (pPSA). METHODS: We established a pPSA-siCXCR4 retrovirus vector and transfected prostate cancer cell PC-3m, LNCaP and breast cancer cell MCF-7, respectively. The expression of CXCR4 mRNA and protein was detected by RT-PCR and western blot, and the ability of adhesion, migration, invasion of prostate cancer cells was assessed using Transwell chamber. A metastasizing model using BALB/cA mice with human bone tissue implantation was established too, and transfected prostate cancer cells were via caudal vein. Survival time of mice suffering bone metastatic tumor as well as the weight and volume of these tumors were recorded and analyzed. RESULTS: The expression of CXCR4 mRNA and protein in androgen-responsive LNCaP cells was blocked by the pPSA-siCXCR4 vector, but it could not work in non androgen-responsive PC-3m cell and breast cancer cell MCF-7. The results of experiments in vitro also showed that the adhesion, transendothelial migration and invasive ability of transfected LNCaP cells were impaired, while there was no change in PC-3m and MCF-7 cells after transfection. pPSA-siCXCR4 represented a similar inhibitory effect in fluorescent bone metastasis model of LNCaP cells compared with PC-3m cells. CONCLUSION: These results suggest that the downstream siRNA controlled by PSA promoter in retrovirus system can express selectively in androgen-responsive prostate cancer in vitro and in vivo, and CXCR4 plays an important role in prostate cancer metastasis. We believe that the pPSA-siCXCR4 retrovirus vector is a potential choice in gene therapy for androgen-responsive prostate cancer.