RESUMEN
MicroRNAs (miRNAs) are small, noncoding RNAs of endogenous origin that play an important role in tumor development. Here, we examined the role of miR-152 in supragalottic laryngeal carcinoma. The expression of miR-152 was assessed by real-time PCR in tissues from 83 patients with supragalottic laryngeal carcinoma in relation to clinicopathological parameters. Cell viability was assessed by thiazolyl blue assay in Hep-2 cells transfected with miR-152 mimics or a negative control. MiR-152 was significantly downregulated in supragalottic laryngeal carcinoma tissues (t = 12.65, p < 0.001, paired t test), and its expression was correlated with pT stage (χ(2) = 26.88, p < 0.001) and pN stage (z = -3.56, p < 0.001) in patients with supragalottic laryngeal carcinoma. MiR-152 inhibited the proliferation of Hep-2 cells. MiR-152 may serve as a novel prognostic marker in patients with supragalottic laryngeal carcinoma.
Asunto(s)
Proliferación Celular , Neoplasias Laríngeas/genética , Neoplasias Laríngeas/patología , MicroARNs/genética , Anciano , Femenino , Estudios de Seguimiento , Humanos , Masculino , Estadificación de Neoplasias , Pronóstico , ARN Mensajero/genética , Reacción en Cadena en Tiempo Real de la Polimerasa , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Células Tumorales CultivadasRESUMEN
This study aimed to investigate the expression, function, and possible mechanism of Src in the Hep-2 cell line. We used Src-specific small interfering RNA (siRNA) to inhibit the expression of Src in Hep-2 cells. RT-PCR and Western blot were applied to evaluate the expression level of Src after RNA interference, and the MTT assay and flow cytometry were used to observe the expression of PI-3 K and Akt. siRNA can downregulate the expression of Src in Hep-2 cells. Downregulation of Src decreased PI-3 K and Akt expression. We found that Src knockdown inhibits the proliferation of Hep-2 cells and the growth of laryngeal carcinoma in vivo. This study has demonstrated that Src participates in the regulation of apoptosis through the Src/PI-3 K/Akt signaling pathway in the Hep-2 cell line. Silencing of Src by siRNA is a viable approach in laryngeal carcinoma treatment.
Asunto(s)
Neoplasias Laríngeas/genética , Fosfatidilinositol 3-Quinasas/metabolismo , Proteínas Proto-Oncogénicas c-akt/metabolismo , Interferencia de ARN , Transducción de Señal , Familia-src Quinasas/genética , Animales , Apoptosis/genética , Línea Celular Tumoral , Supervivencia Celular/genética , Regulación hacia Abajo , Regulación Neoplásica de la Expresión Génica , Humanos , Immunoblotting , Inmunohistoquímica , Neoplasias Laríngeas/metabolismo , Neoplasias Laríngeas/patología , Ratones Desnudos , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Ensayos Antitumor por Modelo de Xenoinjerto , Familia-src Quinasas/metabolismoRESUMEN
A novel drug named Dasatinib is a highly potent ATP-competitive orally active dual Src/Abl kinase inhibitor with anti-proliferative activity against solid tumors and CML (chronic myeloid leukaemia) cell lines. Dasatinib has been shown to have preclinical activity against human prostate, breast, pancreatic, lung, and head and neck cancer. To determine whether Dasatinib can inhibit the growth of laryngeal squamous cell carcinoma, in the present study, we investigated the antitumor effect of Dasatinib on Hep-2 cells. Hep-2 cells were treated with different concentrations of Dasatinib for different time. Cell proliferation, cell cycle distribution, and cell apoptosis were evaluated using MTT assay, flow cytometry, and fluorescent microscopy. It was found that Dasatinib exhibited significant efficacy in growth inhibition, cell cycle arrest at G0/G1 phase, and apoptosis induction in a dose- and time-dependent manner. Measuring the modulation of regulators in the cell cycle, apoptosis and signal transductions by western blot analysis showed that the effect of Dasatinib was due to suppression of the expression of Bax, Bcl-2, Caspase-3, and Caspase-8. Moreover, in vivo studies were performed in a nude mouse xenograft model, the new prescription (DDP + Dasatinib) was better than DDP alone in terms of therapeutic efficacy. In conclusion, the antitumor effect of Dasatinib on Hep-2 cells was due to the induction of cell cycle arrest as well as apoptosis. The possible mechanisms underlying the action might be attributed to the suppression of Src phosphorylation. This investigation suggests a potential clinical application of Dasatinib for the treatment of laryngeal cancer patients.
Asunto(s)
Apoptosis/efectos de los fármacos , Carcinoma de Células Escamosas/patología , Ciclo Celular/efectos de los fármacos , Proliferación Celular/efectos de los fármacos , Neoplasias Laríngeas/patología , Inhibidores de Proteínas Quinasas/uso terapéutico , Pirimidinas/uso terapéutico , Tiazoles/uso terapéutico , Animales , Protocolos de Quimioterapia Combinada Antineoplásica/farmacología , Western Blotting , Línea Celular Tumoral , Transformación Celular Neoplásica/efectos de los fármacos , Transformación Celular Neoplásica/patología , Cisplatino/farmacología , Dasatinib , Relación Dosis-Respuesta a Droga , Citometría de Flujo , Humanos , Técnicas In Vitro , Masculino , Ratones , Ratones Desnudos , Trasplante de Neoplasias , RatasRESUMEN
This study was carried out to evaluate the effects of Astragalus on human nasopharyngeal carcinoma (NPC) viability and apoptosis and to investigate the mechanism of Astragalus in a NPC cell line (CNE2). Cell viability was measured using the MTT assay. CNE2 cells treated with Astragalus were stained with acridine orange/ethidium bromide and subjected to fluorescence microscopy. Bcl-2, Bax, caspase-3 and -8 were measured by western blotting. Rat NPC cells were used to establish a NPC model. Tumor weight, immune organ index and T lymphocyte subsets were employed to detect the immunoregulatory and antitumor effects of Astragalus after administration. Astragalus was effective in inducing apoptosis in CNE2 cells. Morphological changes associated with cell injury were found. Western analysis showed caspase-3, -8, and Bax protein levels were increased after Astragalus treatment, while the bcl-2 protein level was decreased. Astragalus increased the percentage of CD3(+) , CD4(+) T-lymphocytes, and the ratio of CD4(+) /CD8(+) . Astragalus also restored the immunological effects of DDP-induced immunosuppression. These findings suggest that the immunomodulatory and anticancer effects of DDP + Astragalus were better than those of DDP alone, and Astragalus could inhibit immunosuppression induced by DDP. The combination of CDDP + Astragalus could be developed as an effective chemotherapeutic regimen in the treatment of nasopharyngeal carcinoma.
Asunto(s)
Antineoplásicos/farmacología , Planta del Astrágalo/química , Neoplasias Nasofaríngeas/tratamiento farmacológico , Extractos Vegetales/farmacología , Animales , Apoptosis/efectos de los fármacos , Proteínas Reguladoras de la Apoptosis/biosíntesis , Carcinoma , Citometría de Flujo , Humanos , Masculino , Carcinoma Nasofaríngeo , Neoplasias Nasofaríngeas/inmunología , Neoplasias Nasofaríngeas/metabolismo , Neoplasias Nasofaríngeas/patología , Ratas , Ratas Sprague-DawleyRESUMEN
OBJECTIVE: To explore the mRNA expression of KAI1 gene in laryngeal squamous-cell carcinoma and its clinical significance. METHODS: Fresh laryngeal cancer samples taken from 40 laryngeal carcinoma cases and normal control laryngeal tissues from 9 subjects were examined with semi-quantitative reverse transcription polymerase chain reaction (RT-PCR). RESULTS: Moderate, low and negative expression rates of KAI1 gene mRNA in nine normal laryngeal tissues were 33.3% (3/9), 33.3% (3/9) and 33.3% (3/9), respectively. The high, moderate, low and negative expression rates of KAI1 mRNA in 25 laryngeal cancers without lymph node metastasis were 40.0% (10/25), 28.0% (7/25), 20.0% (5/25) and 12.0% (3/25), respectively. The moderate, low and negative expression rates of KAI1 mRNA in 15 laryngeal cancers with lymph node metastasis were 20.0% (3/15), 26.7% (4/15) and 53.3% (8/15), respectively. The KAI1 mRNA expression in the laryngeal cancers without lymph node metastasis was higher than that in normal laryngeal tissues (P < 0.05). The KAI1 mRNA expression in the laryngeal cancers with lymph node metastasis was lower than that in the laryngeal cancers without lymph node metastasis (P < 0.05). The high, moderate and low expression rates of KAI1 mRNA in 10 highly differentiated laryngeal cancers were 50.0% (5/10), 30.0% (3/10) and 20.0% (2/10), respectively. The high, moderate, low and negative expression rates of KAI1 mRNA in 12 low differentiation laryngeal cancers were 8.3% (1/12), 16.7% (2/12), 16.7% (2/12) and 58.3% (7/12), respectively. The differences of KAI1 mRNA expression between high and low differentiation laryngeal cancers were statistically significant (P < 0.05). CONCLUSION: The decrease of KAI1 mRNA expression may be related to lymph node metastasis and low differentiation of laryngeal squamous-cell carcinoma.
Asunto(s)
Carcinoma de Células Escamosas/genética , Proteína Kangai-1/biosíntesis , Neoplasias Laríngeas/genética , Ganglios Linfáticos/patología , Adulto , Anciano , Carcinoma de Células Escamosas/metabolismo , Carcinoma de Células Escamosas/patología , Femenino , Humanos , Proteína Kangai-1/genética , Neoplasias Laríngeas/metabolismo , Neoplasias Laríngeas/patología , Metástasis Linfática , Masculino , Persona de Mediana Edad , ARN Mensajero/biosíntesis , ARN Mensajero/genética , Reacción en Cadena de la Polimerasa de Transcriptasa InversaRESUMEN
OBJECTIVE: To detect mutations of exon 5 and exon 8 of phosphatase and tensin homology deleted on chromosome10/mutated in multiple advanced cancer1/TGF-beta regulated and epithelial cell-enriched phosphatase (PTEN/MMAC1/TEP1 for short PTEN) gene in laryngeal squamous cell carcinoma (LSCC) and analyze the relationship between the mutation and LSCC. METHODS: Fresh tumor samples from 40 LSCC patients were examined using polymerase chain reaction-single-strand conformation polymorphism (PCR-SSCP) and direct DNA sequencing. RESULTS: The mutation of exon 5 and exon 8 occurred in supra-glottic laryngeal carcinoma and no mutation in the glottic laryngeal carcinoma was found. There were 6 cases of mutation for exon 5, and the mutation rate was 15%. In 2 cases, 1 base pair insertion TT-->TAT in codon 85, while in other two cases, 1 base pair deletion GCA-->GC in codon 86, and the two type mutation may result in frame shift mutation. One case had the above two type mutation simultaneously, which may result in missense mutation; 1 case had 1 base pair insertion TT-->TAT in 85 codon and 1 base pair deletion GCA-->GA, which may result in nonsense mutation. The pathology of 3/6 (50%) cases was low differentiation, and the others (3/6, 50%) were middle differentiation. Five cases had lymphatic metastasis, and the rate of which was 83.3%; the other one had no lymphatic metastasis, and the rate of which was 16.7%. There was only 1 case mutation for exon 8, and the mutation rate was 2.5%. One base pair deletion AAA-->AA in 276 codon as well as 1 base pair insertion CT-->CCT between the 336 codon and 337 codon may result in frame shift mutation. The pathology of the case was low differentiation. CONCLUSION: The codon 85,86 of PTEN gene in exon 5 may be "hot spots" in LSCC and the mutation of PTEN gene may be related with the lymphatic metastasis and middle or low differentiation.