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1.
J Huazhong Univ Sci Technolog Med Sci ; 34(4): 535-541, 2014 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-25135723

RESUMEN

The effects of over-expression of testis-specific expressed gene 1 (TSEG-1) on the viability and apoptosis of cultured spermatogonial GC-1spg cells were investigated, and the immortal spermatogonial cell line GC-1spg (CRL-2053™) was obtained as the cell model in order to explore the function of TSEG-1. We transfected the eukaryotic vector of TSEG-1, named as pEGFP-TSEG-1 into cultured spermatogonial GC-1spg cells. Over-expression of TSEG-1 inhibited the proliferation of GC-1spg cells, and arrested cell cycle slightly at G0/G1 phase. Transfection of TSEG-1 attenuated the transcript levels of Ki-67, PCNA and cyclin D1. In addition, over-expression of TSEG-1 induced early and late apoptosis, and reduced the mitochondrial membrane potential of GC-1spg cells. Moreover, transfection of TSEG-1 significantly enhanced the ratio of Bax/Bcl-2 and transcript levels of caspase 9, and decreased the expression of Fas and caspase 8 in GC-1spg cells. These results indicated over-expression of TSEG-1 suppresses the proliferation and induces the apoptosis of GC-1spg cells, which establishes a basis for further study on the function of TSEG-1.


Asunto(s)
Fase G1/fisiología , Histonas/metabolismo , Fase de Descanso del Ciclo Celular/fisiología , Espermatogonias/metabolismo , Animales , Caspasa 8/biosíntesis , Caspasa 8/genética , Línea Celular , Ciclina D1/biosíntesis , Ciclina D1/genética , Histonas/genética , Antígeno Ki-67/biosíntesis , Antígeno Ki-67/genética , Masculino , Ratones , Antígeno Nuclear de Célula en Proliferación/biosíntesis , Antígeno Nuclear de Célula en Proliferación/genética , Espermatogonias/citología , Proteína X Asociada a bcl-2/biosíntesis , Proteína X Asociada a bcl-2/genética
2.
Zhonghua Nan Ke Xue ; 20(6): 495-9, 2014 Jun.
Artículo en Zh | MEDLINE | ID: mdl-25029852

RESUMEN

OBJECTIVE: To explore the mechanism of miR-124 inhibiting the proliferative activity of prostate cancer PC3 cells. METHODS: Luciferase reporter gene assay was used to examine the specific binding ability of miR-124 to PKM2 mRNA 3'-UTR. After miR-124 was transfected mimic to PC3 cells, the expression levels of PKM2 mRNA and protein were detected by real-time fluorescence quantitative PCR (qRT-PCR) and Western blot, respectively. The effects of miR-124 mimic and PKM2 siRNA on the proliferative activity of the PC3 cells were determined by MTT assay. RESULTS: The expressions of PKM2 mRNA and protein were upregulated (5.12 +/- 0.35) times and (4.05 +/- 0.20) times respectively in the PC3 cells as compared with those in the RWPE-1 cells (P < 0.05). Luciferase reporter gene assay demonstrated that miR-124 targeted PKM2 3'-UTR. At 24 hours after transfection with miR-124 mimic, the PKM2 protein expression in the PC3 cells was downregulated (0.16 +/- 0.04) times (P < 0.05), while the PKM2 mRNA level was not changed significantly (P > 0.05), as compared with the control group. MTT assay showed that both miRNA-124 mimic and PKM2 siRNA could inhibit the proliferation of the PC3 cells, but the former exhibited a greater inhibitory effect than the latter. After transfection with miR-124 mimic and PKM2 siRNA, the cell growth rates were (66.20 +/- 5.10)% vs (82.10 +/- 6.35)% at 24 hours (P < 0.05) and (49.34 +/- 2.37)% vs (70.10 +/- 5.80)% at 48 hours (P < 0.05). CONCLUSION: miR-124 can suppress the proliferation of PC3 cells by regulating the PKM2 gene.


Asunto(s)
Proteínas Portadoras/genética , Proteínas de la Membrana/genética , MicroARNs/genética , Neoplasias de la Próstata/patología , Hormonas Tiroideas/genética , Proteínas Portadoras/metabolismo , Línea Celular Tumoral , Proliferación Celular/genética , Humanos , Masculino , Proteínas de la Membrana/metabolismo , Neoplasias de la Próstata/genética , Neoplasias de la Próstata/metabolismo , Hormonas Tiroideas/metabolismo , Transfección , Proteínas de Unión a Hormona Tiroide
3.
Zhonghua Yi Xue Za Zhi ; 93(22): 1740-2, 2013 Jun 11.
Artículo en Zh | MEDLINE | ID: mdl-24124684

RESUMEN

OBJECTIVE: To evaluate the safety and efficacy of preoperative computed tomography urography (CTU) three-dimensional reconstruction, intraoperative radiology and ultrasound guidance followed by percutaneous nephrolithotomy (PCNL) in the treatment of complex renal calculi. METHODS: We summarized the clinical data of 210 patients with complex renal calculi treated at our hospital from December 2008 to December 2011 in this retrospective study. In the one-stop diagnosis and treatment group (n = 119), the optimal puncture approach was designed according to CTU imaging and three-dimensional reconstruction. Percutaneous track was established by ultrasound and radiology guided puncture. PCNL was performed with EMS system. The control group (n = 91) underwent PCNL without radiological guidance. The success rate of puncture, mean accessing time, mean operative duration, intraoperative volume of blood loss and stone-free rate after one operative session were observed. Post-operative follow-ups were conducted until June 2012. RESULTS: Compared to the control group, the one-stop diagnosis and treatment group showed a higher success rate of puncture [98.3% (117/119) vs 92.3% (84/91), P = 0.037], a shorter operative duration [97.8 ± 13.20 vs 110.0 ± 14.73 min, P = 0.043] and a higher stone-free rate after one operative session [92.4% (110/119) vs 83.5% (76/91), P = 0.037]. No significant difference was detected in the mean accessing time[15.3 ± 3.7 vs 13.9 ± 3.9 min, P = 0.398] or intraoperative volume of blood loss [195.8 ± 84.15 vs 263.3 ± 82.06 ml, P = 0.059]. No severe complications occurred. No recurrence of calculi was noted during the follow-up period. CONCLUSION: One-stop diagnosis and treatment plan (CTU 3-D reconstruction plus radiology, ultrasound guidance followed by PCNL) may identify the puncture path, improve the successful rate of puncture and stone-free rates and reduce the complications of PCNL.


Asunto(s)
Cálculos Renales/cirugía , Nefrostomía Percutánea/métodos , Adulto , Anciano , Femenino , Humanos , Imagenología Tridimensional , Cálculos Renales/radioterapia , Masculino , Persona de Mediana Edad , Estudios Retrospectivos , Tomografía Computarizada por Rayos X , Resultado del Tratamiento , Urografía , Adulto Joven
4.
Zhonghua Nan Ke Xue ; 19(2): 102-6, 2013 Feb.
Artículo en Zh | MEDLINE | ID: mdl-23441447

RESUMEN

OBJECTIVE: To study the effect of silencing pyruvate kinase M2 (PKM2) on gambogic acid (GA)-induced apoptosis of human prostate cancer PC3 cells. METHODS: Three specific PKM2 siRNAs and one negative control siRNA (si-NC) were transfected into PC3 cells. The silencing effect of PKM2 siRNAs was determined by real-time fluorescence quantitative PCR (qRT-PCR) and Western blot, and the effects of PKM2 siRNA on the vitality and apoptosis of GA-stimulated PC3 cells detected by MTT and AO/EB double staining, respectively. The mRNA and protein levels of c-myc and cyclin D1 were analyzed by qRT-PCR and Western blot, respectively. RESULTS: All the 3 PKM2 siRNAs effectively reduced the mRNA and protein expressions of PKM2, and PKM2 siRNA-1 exhibited the strongest silencing effect. At 24 h after transfection, the expression levels of PKM2 mRNA and protein were reduced by 70% and 85%, respectively (P < 0.05). Twenty-four hours of treatment with GA (0.5 micromol/L) following transfection with PKM2 siRNA-1 inhibited the vitality of the PC3 cells by 68%, increased their apoptosis, and significantly down-regulated the mRNA and protein levels of c-myc (50% and 35%) and cyclin D1 (60% and 20%) (P < 0.05). CONCLUSION: Inhibition of PKM2 sensitized PC3 cells to GA-induced apoptosis, suggesting that PKM2 may be a potential therapeutic target for sensitizing human prostate cancer to GA.


Asunto(s)
Apoptosis/efectos de los fármacos , Proteínas Portadoras/genética , Proteínas de la Membrana/genética , Neoplasias de la Próstata/genética , ARN Interferente Pequeño , Hormonas Tiroideas/genética , Xantonas/farmacología , Proteínas Portadoras/metabolismo , Línea Celular Tumoral , Humanos , Masculino , Proteínas de la Membrana/metabolismo , Neoplasias de la Próstata/metabolismo , Neoplasias de la Próstata/patología , Interferencia de ARN , Hormonas Tiroideas/metabolismo , Proteínas de Unión a Hormona Tiroide
5.
Acta Pharmacol Sin ; 33(4): 531-41, 2012 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-22426696

RESUMEN

AIM: To investigate the mechanisms underlying the inhibitory effect of gambogic acid (GA) on TNF-α-induced metastasis of human prostate cancer PC3 cells in vitro. METHODS: TNF-α-mediated migration and invasion of PC3 cells was examined using migration and invasion assays, respectively. NF-κB transcriptional activity and nuclear translocation were analyzed with luciferase reporter gene assays, immunofluorescence assays and Western blots. The ability of p65 to bind the promoter of Snail, an important mesenchymal molecular marker, was detected using a chromatin immunoprecipitation (ChIP) assay. After treatment with Snail-specific siRNA, the expression of invasiveness-associated genes was measured using quantitative real-time PCR and Western blot. RESULTS: GA significantly inhibited the viability of PC3 cells at 1-5 µmol/L, but did not produce cytotoxic effect at the concentrations below 0.5 µmol/L. GA (0.125-0.5 µmol/L) dose-dependently inhibited the migration and invasion of PC3 cells induced by TNF-α (10 ng/mL). Moreover, the TNF-α-mediated activation of phosphatidylinositol-3-OH kinase/protein kinase B (PI3K/Akt) and NF-κB pathways was suppressed by GA (0.5 µmol/L). Furthermore, this anti-invasion effect of GA was associated with regulation of Snail. Snail expression was significantly down-regulated by treatment with GA (0.5 µmol/L) in the TNF-α-stimulated PC3 cells. CONCLUSION: GA inhibits TNF-α-induced invasion of PC3 cells via inactivation of the PI3K/Akt and NF-κB signaling pathways, which may offer a novel approach for the treatment of human prostate cancer.


Asunto(s)
Antineoplásicos Fitogénicos/farmacología , FN-kappa B/inmunología , Invasividad Neoplásica/prevención & control , Neoplasias de la Próstata/patología , Factor de Necrosis Tumoral alfa/inmunología , Xantonas/farmacología , Línea Celular Tumoral , Movimiento Celular/efectos de los fármacos , Garcinia/química , Humanos , Masculino , Invasividad Neoplásica/inmunología , Fosfatidilinositol 3-Quinasas/inmunología , Neoplasias de la Próstata/tratamiento farmacológico , Neoplasias de la Próstata/inmunología , Proteínas Proto-Oncogénicas c-akt/inmunología , Transducción de Señal
6.
Mol Cell Biochem ; 346(1-2): 11-21, 2011 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-20824490

RESUMEN

The ubiquitin specific peptidase 22 (USP22) is a positive regulator of the growth of tumors. However, little is known about the impact of USP22 knockdown on the growth of human bladder cells. In the present study, we designed a series of asymmetric interfering RNAs (aiRNAs) and compared the efficacy of aiRNA and conventional symmetric interfering RNA (siRNA) in the silencing of USP22 expression and the growth of human bladder EJ cells in vitro and in vivo. In comparison with transfection with the USP22-specific siRNA, transfection with 15/21 aiRNA was more potent in down-regulating the USP22 expression and inhibiting EJ cell proliferation in vitro. Furthermore, transfection with 15/21 aiRNA induced higher frequency of EJ cells arrested at the G0/G1 phases, but did not trigger EJ cell apoptosis. Moreover, transfection with either the siRNA or 15/21 aiRNA up-regulated the expression of p53 and p21, but down-regulated the expression of cyclin E and Mdm2 in EJ cells. The up-regulated p53 expression induced by the specific siRNA or aiRNA was abrogated by induction of Mdm2 over-expression. In addition, treatment with the specific siRNA or aiRNA inhibited the growth of implanted human bladder tumors in mice and the aiRNA had more potent anti-tumor activity in vivo. Therefore, our data suggest that knockdown of USP22 expression by the aiRNA may down-regulate the expression of Mdm2 and cyclin E, resulting in the up-regulated expression of p53 and p21 and leading to cell cycling arrest and inhibition of human bladder EJ cell proliferation. Our findings indicate that the USP22-specific aiRNA may be a novel approach for the intervention of human bladder tumors.


Asunto(s)
Silenciador del Gen , Interferencia de ARN , Tioléster Hidrolasas/genética , Neoplasias de la Vejiga Urinaria/patología , Animales , Apoptosis , Secuencia de Bases , Línea Celular Tumoral , Cartilla de ADN , Humanos , Masculino , Ratones , Ratones Endogámicos BALB C , Reacción en Cadena de la Polimerasa , ARN Interferente Pequeño/genética , Ubiquitina Tiolesterasa , Neoplasias de la Vejiga Urinaria/genética
7.
Anticancer Drugs ; 22(9): 853-63, 2011 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-21642838

RESUMEN

Methyl jasmonate (MJ) has recently attracted attention as a promising antitumoral compound because of its highly specific proapoptotic properties in a wide range of malignancies. However, the high doses required to achieve a therapeutic benefit have limited its clinical development. Here, we hypothesize that the family of inhibitor of apoptosis proteins (IAPs) may inhibit MJ-mediated apoptosis in cancer cells. We combined MJ with the IAPs inhibitor, the second mitochondria-derived activator of caspases (Smac) peptide to treat bladder cancer cells. The results showed that the combination of MJ and Smac peptide enhanced the apoptosis-inducing effect in a synergistic manner by releasing and activating IAPs-bounding caspase-3. These findings suggest that the inhibition of IAPs could overcome the resistance of cancer cells to MJ.


Asunto(s)
Acetatos/farmacología , Antineoplásicos/farmacología , Apoptosis/efectos de los fármacos , Supervivencia Celular/efectos de los fármacos , Ciclopentanos/farmacología , Proteínas Inhibidoras de la Apoptosis/metabolismo , Oligopéptidos/farmacología , Oxilipinas/farmacología , Neoplasias de la Vejiga Urinaria/tratamiento farmacológico , Proteína Inhibidora de la Apoptosis Ligada a X/metabolismo , Proteína con Homeodominio Antennapedia , Antineoplásicos/metabolismo , Bisbenzimidazol , Caspasa 3/metabolismo , Caspasa 9/metabolismo , Proteínas de Drosophila , Evaluación Preclínica de Medicamentos , Sinergismo Farmacológico , Colorantes Fluorescentes , Células HEK293 , Humanos , Terapia Molecular Dirigida , Oligopéptidos/metabolismo , Survivin , Células Tumorales Cultivadas , Neoplasias de la Vejiga Urinaria/metabolismo
8.
Zhonghua Nan Ke Xue ; 16(9): 803-6, 2010 Sep.
Artículo en Zh | MEDLINE | ID: mdl-21171264

RESUMEN

OBJECTIVE: To evaluate the safety and effectiveness of endourological techniques in the treatment of benign prostate hyperplasia (BPH) in aged high-risk patients. METHODS: We used endourological techniques in the treatment of 283 BPH patients aged over 70 years and complicated with hydronephrosis, renal failure, heart failure, cerebral infarction, respiratory dysfunction, anemia, diabetes, bladder tumor, or prostate weight over 80 g, TURP (transurethral resection of the prostate) for 112 cases and PKRP (transurethral plasmakinetic resection of the prostate) for the other 171. All the patients were followed up for 1-30 months. RESULTS: In the TURP group, the scores on IPSS and QOL were decreased from 27.5 +/- 2.8, 5.5 +/- 1.0 to 5.8 +/- 1.2, 1.0 +/- 0.5, and the residual urine volume (RUV) from (75.0 +/- 20.0) ml to (8.0 +/- 3.0) ml, but the maximal flow rate (Qmax) increased from (6.5 +/- 2.0) ml/s to (18.5 +/- 1.5) ml/s (P < 0.05), while in the PKRP group, the scores on IPSS and QOL were decreased from 28.2 +/- 2.2, 5.5 +/- 1.0 to 5.4 +/- 1.6, 1.0 +/- 0.5, and RUV from (80.0 +/- 20.0) ml to (7.0 +/- 3.0) ml, and Qmax increased from (6.8 +/- 2.1) ml/s to (20.0 +/- 1.5) ml/s (P < 0.05). There were no statistically significant differences in IPSS, QOL, Qmax and RUV after treatment between the two groups (P > 0.05), but significantly less complications were found in the PKRP than in the TURP group (P < 0.05). CONCLUSION: Endourological treatment, especially PKRP, with comprehensive perioperative preparations, unerring operative skills, well-controlled operation time, and intensive postoperative monitoring and nursing, has the advantages of high safety, less bleeding, fewer complications and definite effectiveness for aged high-risk BPH patients.


Asunto(s)
Hiperplasia Prostática/cirugía , Resección Transuretral de la Próstata/métodos , Anciano , Anciano de 80 o más Años , Humanos , Masculino , Calidad de Vida , Resultado del Tratamiento
9.
Zhonghua Nan Ke Xue ; 15(2): 99-105, 2009 Feb.
Artículo en Zh | MEDLINE | ID: mdl-19323366

RESUMEN

OBJECTIVE: To clone the mouse testis specific gene TSEG-2 via a bioinformatic approach. METHODS: The expressed sequence tags (EST) in the normal mouse testis were obtained from the online EST database ZooDDD. Their highly homologous EST sequences were retrieved through the dbEST database to construct contigs and spliced with the biomedical software Biolign. The corresponding exons and introns within the genome sequences were predicted with the software GeneScan. Primers were designed according to the open reading frame. RT-PCR was applied in cloning the cDNA of the novel gene from the mouse testis tissue and analyzing its expression patterns in the undescended testis and various organ tissues as well as in different developmental stages of the mouse testis. The sequencing results of TSEG-2 underwent bioinformatic analyses. RESULTS: The novel mouse testis gene TSEG-2 was successfully cloned, with full-length sequence of 451 bp. The open reading frame was 267 bp, coding a protein of 88 amino acid residues, and demonstrated to be correct by RT-PCR. The expression of TSEG-2 was high in the mouse testis, regular in the testis cDNA samples of different postnatal days, and down-regulated in the cryptorchidism model. No obvious homology with other mouse cDNA was found for TSEG-2. The GenBank accession number EU079025 was achieved. Function prediction showed that mouse TSEG-2 was probably a soluble non-secretary protein located at chromosome 15qE3, or a nucleoprotein with 2 phosphorylation sites of protein kinase C (PKC) and 1 of casein kinase II (CK2). CONCLUSION: A novel mouse testis specific gene TSEG-2 was successfully cloned, which could be down-regulated by cryptorchidism-inducible 17-beta estradiol. This has prepared the ground for further researches on the biological function and expression regulation of TSEG-2.


Asunto(s)
Proteínas/genética , Proteínas/metabolismo , Testículo/metabolismo , Secuencia de Aminoácidos , Animales , Secuencia de Bases , Clonación Molecular , Etiquetas de Secuencia Expresada , Femenino , Expresión Génica , Masculino , Ratones , Ratones Endogámicos , Datos de Secuencia Molecular , Sistemas de Lectura Abierta , Embarazo , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Análisis de Secuencia de ADN
10.
Acta Pharmacol Sin ; 29(7): 861-9, 2008 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-18565287

RESUMEN

AIM: Recent evidence has indicated that members of natural jasmonates, a family of plant stress hormones, exhibit anticancer activity. The current study was undertaken to investigate the effects of jasmonates on the in vitro growth of human neuroblastomas, one of the most common solid tumors in children. METHODS: Cellular proliferation was measured by 3-(4,5-dimethylthiazol-2-yl)-2,5- diphenyltetrazolium bromide colorimetry and colony formation assay. Apoptosis was detected by Hoechst 33258 staining and flow cytometry. Western blotting was applied to assay gene expression. RESULTS: The administration of natural jasmonates, methyl jasmonate, cis-jasmone, and jasmonic acid to cultured neuroblastoma cell line SH-SY5Y, resulted in a decrease of cell proliferation in a doseand time-dependent manner. However, the in vitro growth of cultured human embryonic kidney (HEK) cell line HEK 293 was not affected by jasmonates. The cell cycles of jasmonate-treated SH-SY5Y cells were arrested at the G2/M phase. The incubation of SH-SY5Y cells with jasmonates resulted in characteristic changes of apoptosis. The anticancer activities of natural jasmonates on SH-SY5Y cells are as follows: methyl jasmonate>cis-jasmone>jasmonic acid. In addition, the expressions of proliferating cell nuclear antigen and N-myc were downregulated by methyl jasmonate. Moreover, methyl jasmonate decreased the expression of the Xlinked inhibitor of apoptosis protein and survivin, critical members of inhibitors of the apoptosis protein family, in SH-SY5Y cells. CONCLUSION: Jasmonates suppress the growth of human neuroblastoma cell line SH-SY5Y via inhibiting cell proliferation and inducing apoptosis, which lays the groundwork for further investigation into the anticancer activities and its mechanisms of natural jasmonates on human neuroblastomas.


Asunto(s)
Antineoplásicos Fitogénicos/farmacología , Proliferación Celular/efectos de los fármacos , Ciclopentanos/farmacología , Jasminum/química , Oxilipinas/farmacología , Animales , Antineoplásicos Fitogénicos/química , Apoptosis/efectos de los fármacos , Ciclo Celular/efectos de los fármacos , Línea Celular Tumoral , Ciclopentanos/química , Humanos , Proteínas Inhibidoras de la Apoptosis , Ratones , Ratones Endogámicos C57BL , Proteínas Asociadas a Microtúbulos/biosíntesis , Proteínas Asociadas a Microtúbulos/genética , Oxilipinas/química , Antígeno Nuclear de Célula en Proliferación/biosíntesis , Proteínas Proto-Oncogénicas c-myc/biosíntesis , Relación Estructura-Actividad , Survivin , Ensayo de Tumor de Célula Madre , Proteína Inhibidora de la Apoptosis Ligada a X/biosíntesis , Proteína Inhibidora de la Apoptosis Ligada a X/genética
11.
Yao Xue Xue Bao ; 43(6): 584-90, 2008 Jun.
Artículo en Zh | MEDLINE | ID: mdl-18822959

RESUMEN

This study is to explore the inhibitory effect of methyl jasmonate on cell proliferation and expression of XIAP and survivin of human neuroblastoma cell line BE(2)-C. After cultivation of 1 - 2 mmol x L(-1) jasmonates with BE (2) -C cells for 6 - 24 h, the growth inhibiting rates of BE (2) -C cells were studied by MTT colorimetry. Cell proliferation was detected by colony formation assay. Cell cycle phases were assayed by propidium iodide staining flow cytometery. Cell apoptosis was inspected by acridine orange-ethidium bromide fluorescent staining, Hoechst 33258 fluorescent staining, and Annexin V-FITC and propidium iodide staining flow cytometry. Expressions of cyclin D1, XIAP and survivin were determined by RT-PCR and real-time RT-PCR. Methyl jasmonate inhibited the growth of BE(2)-C cells in a dose- and time-dependent manner. After addition of 1, 1.5 and 2 mmol x L(-1) of methyl jasmonate for 24 h, the inhibiting rates of cell growth reached 20.6% - 85.5% (P < 0.01), and the IC50 was 1.35 mmol x L(-1). The cell cycles were arrested at S phase. A part of cells presented the characteristic morphological changes of apoptosis. The early apoptotic rates were 13.51%, 17.32%, 24.59% (P < 0.01) and the cell death rates were 29.36% , 54.73% , 75.52% (P < 0.01), respectively. The expression of XIAP and survivin mRNA were downregulated by 18.5% - 68.9% , 22.4% - 48.7% (P < 0.05), respectively, without change in that of cyclin D1. The results indicated that methyl jasmonate could significantly inhibit the growth of BE(2) -C cells through inducing cell cycle arrest and apoptosis, downregulating the expression of XIAP and survivin might be one of its molecular mechanisms of action.


Asunto(s)
Acetatos/farmacología , Apoptosis/efectos de los fármacos , Ciclopentanos/farmacología , Proteínas Asociadas a Microtúbulos/biosíntesis , Neuroblastoma/patología , Oxilipinas/farmacología , Proteína Inhibidora de la Apoptosis Ligada a X/biosíntesis , Antineoplásicos Fitogénicos/farmacología , Línea Celular Tumoral , Proliferación Celular/efectos de los fármacos , Ciclina D1/biosíntesis , Ciclina D1/genética , Relación Dosis-Respuesta a Droga , Regulación hacia Abajo , Humanos , Proteínas Inhibidoras de la Apoptosis , Proteínas Asociadas a Microtúbulos/genética , Neuroblastoma/metabolismo , ARN Mensajero/metabolismo , Fase S , Survivin , Proteína Inhibidora de la Apoptosis Ligada a X/genética
12.
Yi Chuan ; 30(3): 352-8, 2008 Mar.
Artículo en Zh | MEDLINE | ID: mdl-18332006

RESUMEN

The expressed sequence tags (ESTs) of normal mouse testis were obtained from online EST database ZooDDD. Their highly homologous EST sequences were found through the dbEST database to construct contigs, and spliced by the biomedical software Biolign. The corresponding exons and introns within genome sequences were predicted by software GeneScan. According to the open reading frame, the primers were designed. RT-PCR was applied in the cloning of novel gene from mouse testis and analyzing its expression pattern in various mouse tissues. The bioinformatics analysis on the sequencing results of TSEG-1 was conducted. Results indicated that a novel gene TSEG-1 was cloned from 1 668-2 011 kb of mouse X chromosome, with full-length sequence of 510 bp. The open reading frame (ORF) is 336 bp in length and en-codes a deduced amino acid sequence of 111 residues. The molecular weight of TSEG-1 protein is 12.84258 kDa, and its pI is 11.4000. RT-PCR demonstrated the correctness of its ORF. TSEG-1 was distinctively expressed in testis, but not in other tissues of mouse. No obvious homology with other mouse cDNA was found for TSEG-1. The GenBank accession number EU079024 was achieved. It was predicted that TSEG-1 is a kind of transmembrane protein, and the transmembrane domain is from 41 amino acid residue to 61 amino acid residue. Blastn analysis revealed its high homology to human testis-specific gene H2AX. Computational prediction of the 5'-untranslated region of TSEG-1 gene revealed a 680 bp-length promoter region. There are four antigen binding sites and two phosphorylation sites of specific protein kinase in TSEG-1 protein, with subcellular localization in mitochondria. The cloning of mouse testis specific gene TSEG-1 laid a foundation for subsequent research of its biological function and expression regulation.


Asunto(s)
Proteínas/metabolismo , Proteínas/fisiología , Testículo/metabolismo , Animales , Secuencia de Bases , Biología Computacional , ADN Complementario/química , ADN Complementario/genética , Etiquetas de Secuencia Expresada , Masculino , Ratones , Datos de Secuencia Molecular , Sistemas de Lectura Abierta/genética , Regiones Promotoras Genéticas/genética , Proteínas/genética , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa
13.
Zhonghua Nan Ke Xue ; 14(1): 3-6, 2008 Jan.
Artículo en Zh | MEDLINE | ID: mdl-18297801

RESUMEN

OBJECTIVE: To explore a method to isolate and purify the subtype of type A spermatogonial stem cells (SSCs). METHODS: We isolated spermatogonia by discontinuous density gradient centrifugation, sorted c-kit-expressed cells with the fluorescence-activated cell sorter (FACS), observed their ultrastructure by electron microscope, and performed immunohistochemistry to determine the expression of c-kit in the testis. RESULTS: The c-kit positive cells constituted (18.65 +/- 1.69) % of the testis cells that were isolated by density gradient centrifugation, but made up only (3.16 +/- 0.84) % of those that were not (P < 0.01). The rates of recovery and viability of the c-kit positive cells sorted by FACS were (65.90 +/- 1.24)% and (85.60 +/- 1.14)%, respectively. CONCLUSION: With c-kit as the marker, FACS can effectively isolate and purify the subtype of SSCs after preliminarily purified by discontinuous density gradient centrifugation.


Asunto(s)
Separación Celular/métodos , Citometría de Flujo/métodos , Espermatogonias/citología , Animales , Masculino , Microscopía Electrónica , Proteínas Proto-Oncogénicas c-kit/biosíntesis , Ratas , Ratas Sprague-Dawley , Espermatogonias/metabolismo , Espermatogonias/ultraestructura , Células Madre/citología , Células Madre/metabolismo , Células Madre/ultraestructura , Testículo/citología , Testículo/metabolismo
14.
Zhonghua Nan Ke Xue ; 14(10): 911-3, 2008 Oct.
Artículo en Zh | MEDLINE | ID: mdl-19157102

RESUMEN

OBJECTIVE: To evaluate the efficacy and safety of vardenafil in kidney transplant recipients with erectile dysfunction. METHODS: Thirty-nine kidney transplant recipients with erectile dysfunction (ED) and serum creatinine values <2 mg/dl were enrolled in a 4-week randomized, double blind, placebo-controlled study, 19 treated with placebo and 20 with vardenafil. Vardenafil efficacy was assessed with the IIEF questionnaire after 4 weeks of treatment, and its safety appraised by measuring serum creatinine levels, creatinine clearances and cyclosporine concentrations before and after the treatment. RESULTS: IIEF scores improved from 12.6 +/- 3.4 to 26.5 +/- 2.8 (P < 0.01), but renal function and cyclosporine concentrations remained unchanged in the vardenafil-treated patients. Adverse effects were observed in 4 patients: headache in 2, palpitation and flush in 1, and dyspepsia in the other. CONCLUSION: Oral vardenafil therapy has a high efficacy and a low incidence of adverse events for kidney transplant recipients with ED.


Asunto(s)
Disfunción Eréctil/tratamiento farmacológico , Imidazoles/uso terapéutico , Trasplante de Riñón , Inhibidores de Fosfodiesterasa/uso terapéutico , Piperazinas/uso terapéutico , Adulto , Método Doble Ciego , Disfunción Eréctil/etiología , Humanos , Masculino , Persona de Mediana Edad , Diálisis Renal , Sulfonas/uso terapéutico , Triazinas/uso terapéutico , Diclorhidrato de Vardenafil
15.
Zhonghua Zhong Liu Za Zhi ; 29(7): 489-94, 2007 Jul.
Artículo en Zh | MEDLINE | ID: mdl-18069626

RESUMEN

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.


Asunto(s)
Antígeno Prostático Específico/genética , Neoplasias de la Próstata , Interferencia de ARN , ARN Interferente Pequeño/genética , Receptores CXCR4/metabolismo , Animales , Línea Celular Tumoral , Regulación Neoplásica de la Expresión Génica , Vectores Genéticos , Humanos , Masculino , Ratones , Células 3T3 NIH , Invasividad Neoplásica , Plásmidos , Regiones Promotoras Genéticas , Neoplasias de la Próstata/genética , Neoplasias de la Próstata/metabolismo , Neoplasias de la Próstata/patología , ARN Mensajero/metabolismo , Receptores CXCR4/genética , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo , Retroviridae/genética , Transfección
16.
Zhonghua Nan Ke Xue ; 13(10): 890-4, 2007 Oct.
Artículo en Zh | MEDLINE | ID: mdl-17977319

RESUMEN

OBJECTIVE: To observe the effects of hydroxycamptothecin (HCPT) on the apoptosis of prostate cancer cell line PC-3 and to explore the possible mechanism. METHODS: The influence of different concentrations (1 x 10(-1), 1 x 10(-2), 1 x 10(-3), 1 x 10(-4) mg/ml) of HCPT on PC-3 cell proliferation at different time (12, 24, 48 h) was determined by tetrazolium (MTT) assay. The morphologic changes of the apoptotic cells were observed by acridine orange/ethidium bromide dyeing. The DNA of the apoptotic cells was analyzed with agarose gel electrophoresis. The apoptosis rate of HCPT on prostate cancer cells was analyzed by flow cytometry (FCM). RESULTS: The growth of PC-3 was inhibited by HCPT in a time- and dose- dependent manner. The values of IC50 were 6.50 x 10(-2) mg/ ml (12 h), 2.35 x 10(-2) mg/ml (24 h) and 5.31 x 10(-3) mg/ml (48 h) respectively. The typical apoptotic cells under the fluorescence microscope showed budding phenomena and apoptotic bodies. And the DNA ladder was observed in ultraviolet light. FCM analysis showed that the apoptosis rate of PC-3 cells increased with the increasing dose of HCPT, which reached the peak (35.76%) at 1 x 10(-3) mg/ml. CONCLUSION: HCPT could suppress PC-3 cell proliferation significantly by inducing the apoptosis of PC-3 cells. However, the mechanism is yet to be further studied.


Asunto(s)
Apoptosis/efectos de los fármacos , Camptotecina/análogos & derivados , Antineoplásicos/farmacología , Camptotecina/farmacología , Línea Celular Tumoral , Proliferación Celular/efectos de los fármacos , Relación Dosis-Respuesta a Droga , Citometría de Flujo , Humanos , Masculino , Factores de Tiempo
17.
J Huazhong Univ Sci Technolog Med Sci ; 37(6): 886-890, 2017 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-29270748

RESUMEN

Emerging evidence has indicated that circular RNAs (circRNAs) play pivotal roles in the regulation of cellular processes and are found to be aberrantly expressed in a variety of tumors. However, the clinical role of circRNAs in bladder cancer (BC) and the molecular mechanisms have yet to be fully understood. In this study, the clinical specimens were obtained and the expression level of a circRNA BCRC4 was detected by real-time PCR in both BC tissues and cell line. The circular RNA over-expression plasmid was constructed and transfected into BC cells and related cell line. The cell cycles and apoptosis were observed using inverted microscope and flow cytometry. Western blotting was used to compare the relative protein expression of groups with different treatments. It was found that circRNA BCRC4 expression was lower in BC tissues than in adjacent normal tissues. Furthermore, consequences of forced-expression of BCRC4 promoted apoptosis and inhibited viability of T24T and UMUC3 cells, and up-regulated BCRC4-increased miR-101 level, which suppressed EZH2 expression in both RNA and protein levels. In addition, gambogic acid (GA) is a promising natural anticancer compound for BC therapy, and GA treatment increased the BCRC4 expression in T24T and UMUC3 cells in a dose-dependent manner. Altogether, our findings suggest that BCRC4 functions as a tumor suppressor in BC, and mediates anticancer function, at least in part, by up-regulating the expression of miR-101. Targeting this newly identified circRNA may help us develop a novel strategy for treating human BC.


Asunto(s)
Apoptosis/genética , Proteína Potenciadora del Homólogo Zeste 2/genética , Regulación Neoplásica de la Expresión Génica , MicroARNs/genética , ARN Neoplásico/genética , ARN/genética , Neoplasias de la Vejiga Urinaria/genética , Antineoplásicos Fitogénicos/farmacología , Apoptosis/efectos de los fármacos , Ciclo Celular/efectos de los fármacos , Ciclo Celular/genética , Línea Celular Tumoral , Proteína Potenciadora del Homólogo Zeste 2/metabolismo , Humanos , MicroARNs/metabolismo , Plásmidos/química , Plásmidos/metabolismo , ARN/agonistas , ARN/metabolismo , ARN Circular , ARN Neoplásico/metabolismo , Estudios Retrospectivos , Transducción de Señal , Transfección , Neoplasias de la Vejiga Urinaria/metabolismo , Neoplasias de la Vejiga Urinaria/patología , Xantonas/farmacología
18.
APMIS ; 114(12): 893-8, 2006 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-17207090

RESUMEN

The serine protease Omi/HtrA2 is released from mitochondria into the cytosol after apoptotic stimuli, inducing apoptosis in a caspase-independent manner through its protease activity and in a caspase-dependent manner by neutralizing the inhibition of inhibitor of apoptosis proteins (IAPs) on caspases. Alteration of apoptosis is essential for cancer development, and cancer cell death by radiation and chemotherapy is largely dependent upon apoptosis. Thus, analysis of the expression status of Omi/HtrA2, a regulator of apoptosis, in cancer tissues is needed for an understanding of cancer development. In the current study we analyzed the expression of Omi/HtrA2 in 65 prostate cancer, 40 benign prostatic hyperplasia and 10 normal prostate specimens by immunohistochemistry. Omi/HtrA2 mRNA levels of in vivo prostate cancer and benign prostatic hyperplasia samples were also assayed by semiquantitative reverse transcription-polymerase chain reaction. Immunopositivity (defined as > or =30%) was observed for Omi/HtrA2 in most of the prostate cancers, and the positive rate of Omi/HtrA2 was lower in the well-differentiated group than in the poorly and moderately differentiated groups (p<0.005). By contrast, the cells in the normal prostate and benign prostatic hyperplasia groups showed no or only weak expression of Omi/HtrA2. Meanwhile, the Omi/HtrA2 mRNA level of prostate cancer is much higher than that of benign prostatic hyperplasia (p<0.001). Taken together, these results suggest that prostate cancer cells in vivo may need Omi/HtrA2 expression for apoptosis, and that Omi/HtrA2 expression might be involved in prostate cancer development.


Asunto(s)
Proteínas Mitocondriales/biosíntesis , Hiperplasia Prostática/enzimología , Neoplasias de la Próstata/enzimología , Serina Endopeptidasas/biosíntesis , Anciano , Apoptosis/fisiología , Serina Peptidasa A2 que Requiere Temperaturas Altas , Humanos , Inmunohistoquímica , Masculino , Persona de Mediana Edad , Hiperplasia Prostática/patología , Neoplasias de la Próstata/patología , ARN Mensajero/biosíntesis , ARN Mensajero/genética , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Estadísticas no Paramétricas
19.
Zhonghua Nan Ke Xue ; 12(10): 872-5, 2006 Oct.
Artículo en Zh | MEDLINE | ID: mdl-17121012

RESUMEN

OBJECTIVE: To investigate the phosphorylation intensity of MAPK pathway molecular Erk1/2 and the proliferation of prostate cancer cell line PC-3M. METHODS: Flow cytometry and RT-PCR were employed to study the ratio of different cell cycles and phases, respectively, before and after GM-CSF stimulation. Erk1/2 phosphorylation intensity was examined by Western blot simultaneously. RESULTS: The rate of PC-3M cells at S and G2/M stages and the expression intensity of Ki-67 increased after GM-CSF incubation in a dose-dependent manner. The phosphorylation intensity of Erk1/2 increased remarkably after stimulation with GM-CSF. CONCLUSION: The intensification of Erk1/2 phosphorylation is one important molecular mechanism of the proliferation of hormone-independent prostate cancer.


Asunto(s)
Factor Estimulante de Colonias de Granulocitos y Macrófagos/farmacología , Proteína Quinasa 1 Activada por Mitógenos/metabolismo , Quinasas de Proteína Quinasa Activadas por Mitógenos/fisiología , Neoplasias Hormono-Dependientes/fisiopatología , Neoplasias de la Próstata/fisiopatología , Línea Celular Tumoral , Proliferación Celular , Humanos , Antígeno Ki-67/biosíntesis , Masculino , Quinasas de Proteína Quinasa Activadas por Mitógenos/metabolismo , Neoplasias Hormono-Dependientes/metabolismo , Neoplasias Hormono-Dependientes/patología , Fosforilación , Neoplasias de la Próstata/metabolismo , Neoplasias de la Próstata/patología , Transducción de Señal/efectos de los fármacos
20.
J Huazhong Univ Sci Technolog Med Sci ; 36(6): 881-886, 2016 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-27924508

RESUMEN

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.


Asunto(s)
Diuresis , Nefrostomía Percutánea/métodos , Cirugía Asistida por Computador/métodos , Adulto , Anciano , Animales , Creatinina/sangre , Femenino , Hemoglobinas/metabolismo , Humanos , Riñón/cirugía , Masculino , Persona de Mediana Edad , Nefrostomía Percutánea/efectos adversos , Complicaciones Posoperatorias , Cirugía Asistida por Computador/efectos adversos , Porcinos , Ultrasonografía
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