A Preliminary Study Of MiRNA Expression Profile In The Cervical Squamous Cell Carcinoma Metastasis

Objectives:The aim of this study is to explore biological markers and biological targets for diagnosis and treatment for the metastasis of cervical cancer and provides experiment basis for regulating mechanism of cervical squamous cell carcinoma Lymph node metastasis associated miRNA using miRNA gene chip by investigating the differential expression of miRNAs between lymph node positive cervical carcinoma and lymph noda negative cervical carcinoma.Methods:1.Collecting samples from cervical cancer patients who underwent hysterectomy and pelvic lymph node dissection in our hospital, including 6 cases in lymph node positive group and 4 cases in pelvic lymph node negative group.2. Extracting the total RNA, and pick out miRNAs that express more than 2 times fold changing by the specimen miRNA gene chip screening in both pelvic lymph node positive group/negative group.3. Picking out eight differential expressed miRNAs, and verify the results of miRNA chip by real-time fluorescence quantitative RT-PCR technology.4. Predicting target genes of these miRNAs which show significant differential expression by bioinformatics net. Results:1) 176 miRNAs have shown differential expression between lymph nodal positive group and lymph nodal negative group via the use of the GenePix proV6.0 software.48 of these miRNAs are found tumor relevant.26 of the 48 miRNAs are up-regulated and the rest 22 are down-regulated. sixteen miRNAs of which are related with tumor metastasis2) Using Real-time quantitative RT-PCR to detect the up-regulated fold change of miR-490-5p, miR-323-3p, miR-328, and the results are 56.89536,8.180334 and 31.94124.While the down-regulated fold change of miR-31,miR-126,miR-144 are 16.3559,9.71628.32.4675. These results are well consistent with miRNA gene chip and are proved to be reliable.3) Among 6 miRNAs proved by RT-PCR, the target genes of miR-490-5p. miR-328,miR-31,miR-144,miR-126 are associated with tumor metastasis。The same target genes also by various miRNAs regulation, the same miRNAs can also control multiple target genes.Conclusion:1.176 miRNAs have shown differential expression between lymph nodal positive cervical carcinoma and lymph nodal negative cervical carcinoma in our experiment by the way of gene microarray. And find 48 of these miRNAs are found tumor relevant and 16 miRNAs of which are related with tumo metastasis, and clear the cervical squamous cell carcinoma metastasis associated miRNAs expression of spectrum preliminary.2. Many targer genes and pathways regulate miRNAs associated with cervical carcinoma nodal metastasis.