The Research Of The MiRNAs Which Regulate Telomerase Of Head And Neck Squamous Cell Carcinoma

Part ?:Screening and verification of miRNAs for regulation of telomeraseObjective:To screen specificity miRNA for telomerase activity regulation using miRNA microarray.Methods:Telomerase-positive cell lines were chosen such as human laryngeal carcinoma cells (Hep-2), human nasopharyngeal carcinoma cells (CNE) and telomerase-negative cell line human osteosarcoma cells (U-2 OS), human lung epithelial cells (BEAS-2B), total RNA was extracted Hy3 fluorescently labeled, miRCURYTM LNA Array (v.11.0) (Exiqon) and hybridized on the chip, real-time quantitative PCR validation miRNAs chip was used.Results:Hsa-miR-301b, hsa-miR-624, hsa-miR-22, hsa-miR-618, hsa-miR-1267 in telomerase-positive cells were highly expressed, and hsa-miR-512-5p, hsa- miR-647, hsa-miR-194, hsa-miR-135a, hsa-miR-621, hsa-miR-640, hsa-miR-155 *was highly expressed in telomerase negative cells. MiR-512-5P in two cells was differentially expressed.Conclusion:miR-512-5P may be involved in the regulation of telomerase activity.Part II:The mechanism of miRNA-512-5P in telomerase regulationObjective:To investigate miRNA-512-5P mechanism for the regulation of tumor telomerase.Methods:The role of miR-512-5p target genes was from miRNA target database, miR-512-5p plasmid was constructed and transfected into tumor cells, inhibition of the target gene was detected and recognized by way of luciferase target and its role point.Results:After multiple miRNA database analysis, we found that miR-512-5p targeting hTERT gene, was significantly decreased in luciferase activity and its decline was obvious. The hTERT protein expression was down in telomerase-positive cells transfected with miR-512-5p plasmidConclusion:MiR-512-5p expression is related with hTERT protein regulating, thereby by inhibiting telomerase activity.Part III:The effect of miR-512-5p on proliferation and telomere of tumor cellsObjective:To investigate the effect of miR-512-5p on proliferation and telomeres of tumor cells.Methods:miR-512-5p plasmid was transfected into tumor cells, the telomere length of tumor cells apoptosis and cell proliferation were changed.Results:MiR-512-5p can inhibit telomere extension, thereby promoting apoptosis of tumor cells, ultimately leading to tumor cell proliferation which was significantly inhibited.Conclusion:MiR-512-5p could inhibit hTERT protein expression, thereby inhibiting telomere extension and ultimately promote tumor apoptosis while proliferation is limited.