Microarray expression analysis has become one of the most widely used functional genomics tools. Efficient application of this technique requires the development of robust and reproducible protocols. We have optimized all aspects of the process, including PCR amplification of target cDNA clones, microarray printing, probe labeling and hybridization, and have developed strategies for data normalization and analysis.
Cervical carcinoma is known as one of the most lethal and common conditions in women worldwide. Increasing evidence shows that microRNAs (miRs) may be involved in the pathogenesis of cervical carcinoma. This study investigates the correlation between expression of miR-224 in peripheral blood mononuclear cells and both diagnosis and prognosis of cervical carcinoma to clarify the effect miR-224 has on the biological behaviors of the subjected cervical carcinoma cells. Initially, 132 patients diagnosed with cervical carcinoma and 120 healthy subjects were recruited. Peripheral blood expression of miR-224 and PTX3 was detected. A telephone follow-up was performed every 3 months after treatment. The diagnostic value of miR-224 in cervical carcinoma was analyzed using the Receiver Operating Characteristic curve. The effects of both miR-224 and PTX3 on cell proliferation, migration, and invasion were evaluated with an intervention of miR-224 ectopic expression or depletion and PTX3 silencing. The bioinformatics prediction website and dual-luciferase reporter assay revealed PTX3 to be a target gene for miR-224. Moreover, miR-224 was detected as over-expressed, but PTX3 was under-expressed in cervical carcinoma. Additionally, as a diagnostic biomarker, a high miR-224 expression was closely linked with the progression of cervical carcinoma. Both miR-224 overexpression and PTX3 silencing promoted cell proliferation, migration, and invasion, whereas, the aforementioned properties were depressed when miR-224 was inhibited. Altogether, the miR-224 overexpression may be a biological indicator in predicting the progression of cervical carcinoma. Thus, miR-224 inhibition may significantly prevent cervical carcinoma progression by targeting the PTX3 gene.
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