A Preliminary Study Of Gene Expression Profile In Cervical Squamous Cell Carcinoma With Pelvic Lymph Node Metastasis

Explore the differentially expressed gene profile in cervicalsquamous cell carcinoma tissues with and without pelvic lymph node metastasis by the application of microarray technique. To search for potential gene markers for the prediction of pelvic lymph node metastasis and to provide guidelines for the study of metastasis mechanism of cervical squamous carcinoma. Method:1. Cancer tissue samples were collected from4cervical squamouscell carcinoma patients with pelvic lymph metastasis and6patients without metastasis before treatment. All10patients had undertaken radical hysterectomy and pelvic lymphadenectomy in our hospital.2. Total RNA was extracted and cDNA was synthesized by reversetranscription. The differentially expressed gene profile was identified between cervical squamous cell carcinoma tissues with and without pelvic lymph node metastasis by NimbleGen microarray.3. Fluorescent real-time quantitative PCR was employed to validatethe accuracy of the microarray by detecting the expression of6candidate genes picked from the differentially expressed gene profile. The method of2-ΔΔC was taken to express the fold change of the6genes.4. PUBMED and gene ontology database were applied to analyze and classify the function of the differentially expressed genes. Genes associated with tumor metastasis were identified.Results:1. Differentially expressed gene profile:With the application of microarray,329genes were identified as markedly differentially expressed genes between cervial squamous cell carcinoma tissue with and without pelvic lymph node metastasis.187genes were up-regulated and142genes were down-regulated in cervical squamous cell carcinoma tissues with pelvic lymph node metastasis.2. Results of real-time PCR:KLK6, FGA, CTGF, CPB2, PRL and CCL20in the profile were detected by real-time PCR, and the fold change of each was respectively up-regulated2.7014,11.7411,3.4248,12.1232,4.4272and down-regulated12.7065, which was in accordance with the results of microarray.3. Genes associated with tumor metastasis:44genes associated with tumor metastasis were identified,29of which were up-regulated in cervical squamous cell carcinoma tissues with pelvic lymph node metastasis and15ones were down-regulated.Conclusions:1.By the application of gene microarray and real-time PCR. gene profile in cervical squamous cell carcinoma tissues associated with pelvic lymph node metastasis was identified. 2. Quantity of tumor-associated genes existed in differentially expressed gene prolife in cervical squamous cell carcinoma tissues, which indicated that lymph node metastasis of cervical squamous cell carcinoma was a process involving numerous genes.