Basic and Translation Research of miRNA-196 in Head and Neck Cancer.

Head‐neck cancer (HNC) (including oral cancer and larynx cancer) is one of the ten frequent cancers in the world with an estimated over 500,000 new cases being diagnosed annually. In Taiwan, the incidence of head and neck cancer has become the 6th leading cancer, the 4th leading cancer in male since year 2009, and is increasing in the recent years. Since this cancer usually occurs in the middle age male, at the high peak of life responsibility, it has tremendous impact of family and society. MicroRNAs (miRNAs) are a prevalent class of small single‐stranded non‐coading RNAs (19‐25 nts). They serve widespread functions as regulatory molecules in post‐transcriptional gene silencing for the regulation of cellular proliferation, differentiation, and apoptosis, which have great impacts on the malignant transformation. Apparently, the expression of miRNA profile is closely associated with cancer. Previously, we have global surveyed 470 human miRNAs associated with HNC by using microarray method to compare the differential expression levels between 6 cancer cell lines and 5 lines of normal keratinocytes. Total of 23 miRNAs were found significantly different between two groups (P ＜ 0.05), with 19 elevated and 4 decreased in HNC cell line. These miRNA were validated by quantitative RT‐PCR analysis, and miR‐196 family molecules consistently showed over‐expression in the cancer cell lines. In this present study, we intend to further investigate the potential role miR‐196 family in the carcinogenesis of HNC. The specific aims and research designs are briefly describe below. Aim #1. Functional investigation of miR‐196 in the oncogenesis of HNC. Examination of the cellular effects after alteration of the specific miRNA expression in cells. (1) Construction of the miR‐196 antigomir or overexpressed plasmids and transfection into HNC cells. (2) Functional study on cellular homeostatsis, including growth, migration and invasion ability. Aim #2. Mechanistic investigation of miR‐196 signaling pathway. (1) Prediction of miR‐196 downstream target genes by bioinformatic analysis. (2) Determination of the target expressions at both mRNA (by RT‐qPCR) and protein (by western blot) levels. (3) Confirmation by luciferase reporter assay of the target‐3’‐UTR upon alteration of miR‐196 expressions. (4) Functional validation of the target genes, which should be complement with miR‐196 function and rescues miR‐196 effects. Aim #3: Translational investigation of miR‐196 as a diagnostic / prognostic marker. (1) Evaluation of miR‐196 levels in the specimens of cancer tissues, plasma, saliva or oral rinses from HNC patients and non‐cancer subjects. (2) Comparison of miR‐196 expressions between two groups or correlation with the clinicopathological features of HNC patients. Aim #4: Translational investigation of miR‐196 as a therapeutic target. (1) Investigation of the tumor growth and metastatic status after delivery of miR‐196 antigomir into xenografted tumor mice. (2) Examination of tumor growth, metastasis, mice survival and confirming miR‐196 and the target expressions on the xenografted tumor sections.

Project ID:PC10301-0899
External Project ID:NSC101-2320-B182-041-MY3