RESUMO
Ovarian cancer is one of the most common gynecologic malignancies that has a poor prognosis. THUMPD3-AS1 is an oncogenic long noncoding RNA (lncRNA) in several cancers. Moreover, miR-320d is downregulated and inhibited proliferation in ovarian cancer cells, whereas ARF1 was upregulated and promoted the malignant progression in epithelial ovarian cancer. Nevertheless, the role of THUMPD3-AS1 in ovarian cancer and the underlying mechanism has yet to be elucidated. Human normal ovarian epithelial cells (IOSE80) and ovarian cancer cell lines (CAVO3, A2780, SKOV3, OVCAR3, and HEY) were adopted for in vitro experiments. The functional roles of THUMPD3-AS1 in cell viability and apoptosis were determined using CCK-8, flow cytometry, and TUNEL assays. Western blot was performed to assess the protein levels of ARF1, Bax, Bcl-2, and caspase 3, whereas RT-qPCR was applied to measure ARF1 mRNA, THUMPD3-AS1, and miR-320d levels. The targeting relationship between miR-320d and THUMPD3-AS1 or ARF1 was validated with dual luciferase assay. THUMPD3-AS1 and ARF1 were highly expressed in ovarian cancer cells, whereas miR-320d level was lowly expressed. THUMPD3-AS1 knockdown was able to repress cell viability and accelerate apoptosis of OVCAR3 and SKOV3 cells. Also, THUMPD3-AS1 acted as a sponge of miR-320d, preventing the degradation of ARF1. MiR-320d downregulation reversed the tumor suppressive function induced by THUMPD3-AS1 depletion. Additionally, miR-320d overexpression inhibited ovarian cancer cell viability and accelerated apoptosis, which was overturned by overexpression of ARF1. THUMPD3-AS1 inhibited ovarian cancer cell apoptosis by modulation of miR-320d/ARF1 axis. The discoveries might provide a prospective target for ovarian cancer treatment.
Assuntos
Fator 1 de Ribosilação do ADP , Apoptose , Regulação Neoplásica da Expressão Gênica , MicroRNAs , Neoplasias Ovarianas , RNA Longo não Codificante , Feminino , Humanos , Fator 1 de Ribosilação do ADP/metabolismo , Fator 1 de Ribosilação do ADP/genética , Apoptose/genética , Linhagem Celular Tumoral , Proliferação de Células , MicroRNAs/genética , MicroRNAs/metabolismo , Neoplasias Ovarianas/genética , Neoplasias Ovarianas/patologia , Neoplasias Ovarianas/metabolismo , RNA Longo não Codificante/genética , RNA Longo não Codificante/metabolismo , RNA Antissenso/genéticaRESUMO
OBJECTIVE: To investigate the expression of mesothelin (MESO) mRNA and protein and its significance in ovarian carcinomas. METHODS: Semi-quantitative reverse transcription polymerase chain reaction (RT-PCR) and immunohistochemistry were used to detect the expression level of MESO mRNA and protein, respectively, in 124 samples of ovarian tumor and normal tissues, including 84 epithelial ovarian carcinomas, 12 borderline ovarian tumors, 16 benign ovarian tumors and 12 normal ovarian tissues. RESULTS: The expression of MESO mRNA and protein in epithelial ovarian carcinomas (1.4005 +/- 0.4646, 2.7857 +/- 2.2712) and borderline ovarian tumors (1.0650 +/- 0.3100, 2.9167 +/- 2.391) were significantly higher than that in benign ovarian tumors (0.6463 +/- 0.2419, 1.2500 +/- 1.6125) and normal ovarian tissues (0.6439 +/- 0.2729, 0.9167 +/- 1.2401) (P < 0.05), and also significantly higher in serous cystadenocarcinoma (1.5255 +/- 0.4151, 3.3036 +/- 2.6141) and endometrioid carcinoma (1.5250 +/- 0.5419, 3.0000 +/- 2.3094) than that in mucinous cystadenocarcinoma (1.0675 +/- 0.3149, 1.0556 +/- 1.9242) (P < 0.05). The expression of MESO mRNA and protein in stages II and IV carcinomas (1.5100 +/- 0.4142, 3.6087 +/- 3.3959) was significantly higher than that in stages I and II carcinomas (1.1190 +/- 0.4909, 1.7895 +/- 2.6320; P < 0.05), and also significantly higher in grade 3 carcinomas than that in grade 1 and 2 ones (P < 0.05), but was not correlate with age or serum CA125 of the patients (P > 0.05). CONCLUSION: The results of this study demonstrated that the expression of MESO mRNA and protein is increased in ovarian carcinomas and borderline ovarian tumors, and MESO may play a role in the adhesion and dissemination of ovarian carcinomas.
Assuntos
Carcinoma Endometrioide/metabolismo , Cistadenocarcinoma Mucinoso/metabolismo , Cistadenocarcinoma Seroso/metabolismo , Glicoproteínas de Membrana/metabolismo , Neoplasias Ovarianas/metabolismo , Carcinoma Endometrioide/genética , Carcinoma Endometrioide/patologia , Estudos de Casos e Controles , Cistadenocarcinoma Mucinoso/genética , Cistadenocarcinoma Mucinoso/patologia , Cistadenocarcinoma Seroso/genética , Cistadenocarcinoma Seroso/patologia , Feminino , Proteínas Ligadas por GPI , Regulação Neoplásica da Expressão Gênica , Humanos , Imuno-Histoquímica , Mesotelina , Pessoa de Meia-Idade , Metástase Neoplásica , Estadiamento de Neoplasias , Neoplasias Ovarianas/genética , Neoplasias Ovarianas/patologia , Ovário/metabolismo , Ovário/patologia , RNA Mensageiro/metabolismo , Reação em Cadeia da Polimerase Via Transcriptase ReversaRESUMO
IL-6 has been found to be associated with poor response to chemoradiotherapy and poor overall prognosis of patients with cervical cancer. However, little is known about the clinicopathological significance of IL-6 receptor (IL-6R) expression in the setting of cervical cancer. To investigate the clinicopathological meaning of IL-6R in cervical cancer, expression of IL-6R was detected using immunohistochemistry in cervical cancer tissue microarray composed of 98 cases of cervical cancer and paired normal controls. As further confirmation of expression trend, western-blotting was conducted in another independent 36 pairs of cervical cancer and matched normal controls. Subsequently, the statistical correlation between IL-6R expression and clinicopathological variables was analyzed, including demographic, TNM stage, clinical grading and overall prognosis. IL-6R expression was shown to be remarkably associated with lymph node metastasis, recurrence and overall prognosis. Moreover, only IL-6R expression was observed to be an independent prognostic factor among these variables that could potentially influence the overall prognosis of patients with cervical cancer. In conclusion, IL-6R was shown to be an independent prognostic factor for patients with cervical cancer.