人elac2基因在肿瘤中的生物学功能分析-biological function analysis of human elac 2 gene in tumor.docx

Abs Abstract Study on Biological Functions of Human ELAC2 Gene IV IV V V Study on Biological Functions of Human ELAC2 Gene Abstract Objective The current study was firstly to investigate the effect and the mechanism of ELAC2 gene on the growth, migration and invasion as well as radiosensitivity in human breast cancer and prostate cancer cell lines. Secondly, the study was to illucidate the functions of ELAC2 in mediating androgen receptor transcriptional activity via a direct interaction with androgen receptor. These novel studies will first empirical evidence for the potential application of ELAC2 as a gene therapy in breast cancer and prosate cancer in the near future. Methods A recombinant mammalian plasmid pcDNA3 carrying a full-length of ELAC2 cDNA (pcDNA3-ELAC2) was transfected into two breast cancer cell lines, MDA-MB-231 and MCF-7, and one prostate cancer cell line, DU-145, via Lipofectamine transfection assay. Pooled G418-resistant clones were obtained after incubation in G418-containing medium for three weeks. Alterations of ELAC2 mRNA and protein level were monitored by RT- PCR and Western blot assay. Cell proliferation was assessed by MTT assay. Flow cytometric analysis was used to detect the cell cycle distribution and apoptosis induction. Wound healing assay and modified Boyden chamber assay were performed to assess the ability of cell migration and invasion. Western blot assay was employed to evaluate the alterations of proteins. The allograft tumor model was established in BALB/c nude mice via subcutaneously inoculation with different cancer cells, and the effects of ELAC2 on tumor growth, invasion and metastasis were examined. The amount of microvessels in tumors was measured by immunohistochemistry and compared afterward. The metastasis model was established by injecting cancer cells into nude mice through vena caudalis. Study on Biological Functions of Human ELAC2 Gene Colony formation assay was used to evaluate survival fraction after irradiatio