Study On The Regulation Of UNC5A On The Biological Behavior And Radiosensitivity Of Non-Small Cell Lung Cancer

Abstract

UNC5A has been proved down regulated in many kinds of neoplasms. UNC5 family is currently identified as tumor suppressor genes. In this study, we investigated UNC5A expression in NSCLC and its function as a prognosis predictor and a biological regulator. 1. Using database analysis, the expression of UNC5A in NSCLC tumor and para-tumor tissue. Comparing the UNC5A promotor region methylation of tumor and para-tumor and analyze the relationship between the methylation and clinical pathology characters. Using BSP to assess UNC5A methylation level and its expression in NSCLC. Colony formation to investigate radiosensitivity after demethylated. 2. Using database analysis, the relationship between UNC5A expression and prognosis. IHC staining was performed on pre-radiotherapy cases and the relationship between UNC5A expression and pathological characters and prognosis was explored. Real-time PCR was adopted on NSCLC cases and cell lines to analysis pathological characters comparing with para-tumor tissue. 3. Building UNC5A-silenced cell lines and over-expression cell lines then we performed MTT to assess proliferation, Transwell to investigate invasive ability, scratch test to investigate migration, flow cytometry test apoptosis ability. Colony formation to investigate radiosensitivity. Using Western bolt to assess AKT/PhosphoAKT, PI3K/PhosphoPI3K, p21, and cyclin D1 expression. 1. UNC5A expression is higher in tumor tissue than normal tissue. The methylation level of tumor tissue was higher than para-tumor and associated with clinical staging. Methylation of UNC5A promotor region was also increased in NSCLC cell lines compared with normal cell line. UNC5A methylation was decreased and mRNA was increased, also increased radiosensitivity of tumor cells after demethylated. 2. High expression of UNC5A predicts better prognosis in NSCLC database. IHC staining indicated that UNC5A expression is negatively related to TNM staging. UNC5A high expression suggested better outcome after radiotherapy. UNC5A expression was significantly negatively related to TNM staging and tumor size. 3. UNC5A silencing increased tumor proliferation, invasive ability and migration, while over-expression of UNC5A decreased these characters in vitro. Over-expression of UNC5A increase radiosensitivity. UNC5A expression was related to PI3K/AKT/P21 signaling pathway and cyclin D1 in NSCLC cell line. 1. In NSCLC tumor tissue, methylation of UNC5A promotor region was much higher than para-tumor tissue and related to TNM staging and associated with radiosensitivity.