The Experimental Research Of MiRNAs In The Regulation Of OSCC Cell Proliferation And Apoptosis

Background & Purpose: Head and neck squamous cell carcinoma(HNSCC) has been a serious problem with more than 500,000 incidence and caused 200,000 deaths worldwidely every year. Despite huge progress has been made in diagnosis and treatment of HNSCC, it is still a great threat to human life with an unsatisfied 5-year survival. Micro RNAs(mi RNA) are small noncoding single-strand nucleotide(19-23nts) that regulate protein-coding gene expression in a sequence-specific manner and thereby influence cell fate and function. Tumor is the result of genetic mutations of, it is accompanied by a variety of mi RNAs in the process of the formation and development of abnormal expression of this abnormal expression with tumor site and organization change and pathological characteristics. In order to more in-depth understanding of mi RNAs in oral squamous cell carcinoma of the role played by, this research to influence regulation of oral squamous cancer cell proliferation and apoptosis of mi RNAs screening, identification and further experiments have been carried out to verify the specific mechanism of action.Methods: We choose 8 pairs of samples of oral squamous cell carcinomas and adjacent normal tissues for microarray analysis to screen differentially expressed mi RNAs in oral squamous carcinoma tissues of mi RNAs; In clinical samples and the department of oral squamous cancer cells by using the method of real- time PCR for difference, in vitro cell experiment method to screen out the differentiation of mi RNAs to detect the gene function. We conducted in vitro cell proliferation test(CCK8) and single cell cloning experiment, apoptosis and cell cycle detection, in the evaluation of screening of mi RNAs effects on oral squamous cancer cell proliferation and apoptosis. Through the analysis and prediction of bioinformatics website and dual luciferase reporter gene system to find and verify the relevant target genes; Application of transcript copy number detection and Western blot method(Western blot) validation of mi RNAs to the appropriate target genes in the m RNA and protein level control function.Result: mi R-330-3p in 29 patients with oral squamous cell carcinomas clinical tissue samples(each pair of samples including oral squamous carcinoma tissues and the normal tissue adjacent to carcinoma) expressed as: in the tissue adjacent to carcinoma in the expression of mi R-330-3p is 7.476 ± 3.537(10-3 × U6), the expression of Oral Squamous Carcinoma is 13.059 ± 5.045(t=5.282,n=29,P<0.0001).The the two groups have statistical significance. mi R-330-3p in proliferation ability weaker HN4 and HN6 cell line in the low expression level, and in high proliferation HN13 and SCC25 expression level is higher; mi R-330-3p expression can obviously improve the ability of cancer cells proliferation and single cell clone formation, and significantly reduce the apoptosis rate of oral squamous cancer cells; Research found that mi R-330-3p to negative regulating target genes m RNA and protein level factor 4(PDCD4) the expression of programmed cell apoptosis, reduce the apoptosis rate of squamous cancer cells, thus indirectly promote the proliferation of tumor cells.Conclusions: mi R-330-3p expression in oral squamous cancer cells is significantly higher than normal tissue adjacent to carcinoma, and negatively correlated with the prognosis of patients with; mi R-330-3p can bind with 3’UTR, regulating the expression of suppressor PDCD4 negatively, significantly reduce the apoptosis rate of oral squamous cancer cells, play the role of promoting cancer gene.