| Background The pathological type of ovarian cancer is complicated, which contains majority of epithelial cancers, mainly including three subtypes such as serous carcinoma, endometrioid carcinoma and mucinous carcinoma. Serous ovarian cancer accounts for the most, with biological behaviors of occult early symptoms, easy to metastasize and widely disseminated, so more than 70% of patients visit doctors at advanced stage, and five-year survival is only 30%. Ovarian cancer, with the mortality ranks first in gynecological malignancies, has become a major serious threat to women’s life and health, due to the lack of early diagnosis methods of ovarian cancer. Despite a lot of effort was made in the medical researches and clinical studies, ovarian cancer survival rates of nearly fifty years did not change significantly. However, the pathogenesis of ovarian cancer is still unclear, therefore, it will be very significant to find its closely related genes about genesis and development, and to block its key occurrence.Recently the early high-grade serous ovarian cancer lesions, which are called serous carcinoma in situ, discovered at the site of tubal fimbria of fallopian tube, providing new concepts and materials for the study of early stage of ovarian cancer. Recent studies have found that micro RNA(mi RNA) can affect cell proliferation, apoptosis, migration and invasion, and other functions by regulating the expression of some tumor suppressor genes and oncogenes. Many studies have found that mi RNA expression profiles are abnormal in a variety of tumor tissue, suggesting that the expression of this disorder is potentially relevant with the genesis and development of tumor. mi RNA also plays an important role in ovarian cancer. Our previous study found that mi R-129-1-3p/mi R-129-2-3p in serous ovarian carcinoma was significantly abnormal in expression using mi RNA microarray technology by detecting the eight pairs of serous ovarian cancer tissues and normal tubal fimbria tissues. Studies found that mi R-129 plays an important role in occurrence and development of gastric cancer and liver cancer, however, its role and possible mechanism in epithelial ovarian cancer, has been not reported so far. In this study, further experiments were performed to detect the expression of mi R-129-1-3p/mi R-129-2-3p in serous ovarian cancer and take an insight into its biological function and possible mechanism in ovarian carcinogenesis and development.Objective 1. Identify the expression levels of mi R-129-1-3p/mi R-129-2-3p in serous ovarian carcinoma. 2. Identify the role of mi R-129-1-3p/mi R-129-2-3p in regulation functions of serous ovarian cancer cell line Skov3 cell proliferation, cell cycle, migration and invasion. 3. Preliminarily explore mechanism of mi R-129-1-3p/mi R-129-2-3p functioning in serous ovarian cancer.Methods 1. Total RNA was extracted from the 100 cases of serous ovarian cancer tissues in patients after surgical resection and 50 cases of normal tubal fimbria tissues. After reverse transcription, the relative expression of mi R-129-1-3p/mi R-129-2-3p in cancer tissues and normal tissues was detected by q RT-PCR and their differential expression level was analyzed by relative quantitative methods. 2. The human serous ovarian cancer cell lines Skov3 were transiently transfected by mi R-129-1-3p/mi R-129-2-3p-mimics with Hi Per Fect reagent. And the functions of cell migration, invasion and proliferation after overexpression of mi R-129-1-3p/ mi R-129-2-3p were detected througth scratch, transwell migration, invasion and MTT assay. 3. The cell invasion, migration-related oncogenes which will be the mi R-129-1-3p/mi R-129-2-3p possible target genes was selected by bioinformatics analysis using software DIANA LAB, Target Scan and Mi Randa and the DAVID database application functional cluster analysis. Its predicted target genes proteins were detected by Western-blot method for its regulated expression in ovarian cancer cells within Skov3 overexpression of mi R-129-1-3p/mi R-129-2-3p. And the regulating relationship between mi R-129-1-3p/mi R-129-2-3p and their candidate target genes was verified by reporter gene experiments with dual luciferase.Results 1. The expression of mi R-129-1-3p/mi R-129-2-3p was significantly decreased in serous ovarian carcinoma tissues compared with normal tubal fimbria tissues. 2. Skov3 cells migration was significantly weakened after transiently transfected by mi R-129-1-3p/mi R-129-2-3p-mimics with Hi Per Fect reagent through scratch test and transwell experiment. Skov3 cells proliferation was significantly weakened after overexpression of mi R-129-1-3p/mi R-129-2-3p through MTT experiments. Transwell invasion assay suggest that tumor cells invasion capacity significantly weakened afteroverexpression of mi R-129-1-3p/mi R-129-2-3p, relatively. 3. BZW1 was possible target gene of mi R-129-1-3p/mi R-129-2-3p by the bioinformatics forecast. And BZW1 expression was significantly decreased in ovarian cancer Skov3 cells after overexpression of mi R-129-1-3p/mi R-129-2-3p through q RT-PCR and Western-bolt experiments. BZW1 was proved to be the target gene of mi R-129-1-3p/mi R-129-2-3p by dual luciferase reporter gene experiment.Conclusions 1. The expression of mi R-129-1-3p/mi R-129-2-3p was significantly decreased in serous ovarian carcinoma. In vitro, overexpression of mi R-129-1-3p/mi R-129-2-3p could significantly inhibit ovarian cancer cell migration, proliferation and invasion. The mi R-129-1-3p/mi R-129-2-3p may be a tumor suppressor gene and restrain ovarian cancer invasion and metastasis. 2. BZW1 is one of mi R-129-1-3p/mi R-129-2-3p possible target genes, suggesting that it may play suppression of ovarian cancer invasion and metastasis by inhibiting the expression of BZW1. In conclusion, this preliminary study showed that mi R-129-1-3p/mi R-129-2-3p may be new candidate tumor suppressor genes in ovarian cancer, and their mechanism may be carried out by regulating the expression level of BZW1. It provides a new strategery and primary theory basis to clarify the mechanisms of the occurrence and progression of ovarian cancer, and for further gene therapy and individualized diagnosis and therapy. |
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