ShRNA-hIAP2Enhancing Radiation Effect On Nasopharyngeal Carcinoma Cell

NPC is one of common malignant tumors, originating in the nasopharyngeal mucosa lining epithelium, malignant conversion is high, natural survival was18.7months on average, onset hidden, difficult to detect early. China Guangdong, Guangxi, Fujian, Hunan and other places of multiple areas, more men than women. Age of onset mostly middle-aged, some young ill. Etiology and racial predisposition, yellow and more sick than Caucasians, and genetic factors and EB virus infection and so on, can occur early cervical lymph node metastasis of nasopharyngeal cancer patients between the natural course of the disease varies greatly. From the onset of symptoms to death from the natural course of3to13months. Currently, the main treatment with radiation therapy. Seen from the existing domestic and international coverage, nasopharyngeal carcinoma after radiotherapy5-year survival rate of8%to62%. With radiation therapy equipment upgrades, radiation therapy technology improvements, Radiotherapy after5-year survival rate continues to increase. Report a Shanghai Tumor Hospital,1955previously applied deep X-ray therapy,5-year survival rate was8%in1983,5-year survival rate was54%. After radiotherapy for local recurrence and distant metastasis is the main reason for death of the patient, so in addition to improving radiotherapy technology to improve radiotherapy, but also on the biological characteristics of nasopharyngeal carcinoma, nasopharyngeal carcinoma patient body aspects as well as the interaction between tumor and patient factors such as body studied. According to the biological characteristics of nasopharyngeal carcinoma patients body, in treatment from radiotherapy, chemotherapy, surgery, immunotherapy, gene therapy, traditional Chinese medicine and other treatment methods considered, selection and development of appropriate treatment program to further improve the efficacy. In this study, nasopharyngeal carcinoma cell line CNE1as the research object, construct shRNA-hIAP2, using RNA interference as a means by liposome-mediated transfection methods, research MAP2CNE1cell lines for enhanced radiosensitivity of the initial investigate the mechanism for the adjuvant treatment of clinical gene lay a certain foundation.Abstract:Objective:Construct the inhibitor of the shRNA of apoptosis protein (hIAP-2) for the purpose of the combination of inhibition hIAP-2gene with radiotherapy, investigate its effect towards CNE1nasopharyngeal carcinoma cell line radiosensitivity.Methods:First, using the MTT assay to determine the optimal dose of radiation; Then build four hIAP-2-shRNA silencing fragment and transfected by liposome CNEl cells, and finally use real-time quantitative PCR (Q-PCR) and Western blot (WB, Western blot) best silencing shRNA sequences screened; Using silence shRNA sequences best transfection CNE1cells, respectively set up transfected irradiated group and non-transfected irradiated group; using real-time quantitative PCR and WB detect hIAP-2gene and protein expression, using immunocytochemistry detect hIAP-2protein expression,apoptosis detectes by flow cytometry, transwell measures cell invasion.Results:MTT assay determined4Gy as the optimal radiation dose; Q-PCR and WB test results showed that four hIAP-2-shRNA could effectively inhibit hIAP-2mRNA and protein expression, which hIAP-2-shRNA2silence has the most significant effect; Q-PCR,WB and immunocytochemistry test results showed that hIAP-2gene and protein expression of the transfection hIAP-2-shRNA2irradiated group and untransfected hIAP-2-shRNA2CNE1cells irradiated group were significantly different; Flow cytometry showed that transfection hIAP-2-shRNA2irradiated group than in the non-transfected hIAP-2-shRNA2irradiation apoptosis rate were significantly different; Transwell test results showed that transfection hIAP-2-shRNA2irradiated group and untransfected hIAP-2-shRNA2invasion irradiated group were significantly different;Conclusion:Transfection hIAP-2gene silencing after radiotherapy can increase nasopharyngeal carcinoma cell apoptosis rate, reducing its invasion, can play a role in radiosensitization.