Investigation On The Gene Expression Pattern In Cervical Squamous Cell Carcinoma With CDNA Microarray

Objective:To investigate the differentially expressed gene in cervicalsquamous cell carcinoma use DNA microarry , explore the significanceof DNA microarry in expediting our understanding of the expressionpattern of cervical cancer. Method:(1)Collecting and dealing with the samples : The tissueswere collected from 3 samples of cervical cancer in Ib and 3 samples ofuterine leiomyoma during surgery, the tissues were put into fluid nitrogenimmediately and kept there. The histology types of the tissues wereapproved by pathology examination. (2) Picking up mRNA and judging thequality of the mRNA: The mRNA were isolated and purified from samplesof cervical cancer and normal tissues and checked with the instrumentnamed Agilent. The mRNA were proved with high quality. (3) preparinghybridization probes: The mRNA were reversely transcripted into cDNAwith the incorporation of fluorescent Cy-3dCTP or Cy-5dCTP from theother to prepare two groups of hybridization probes. The samples ofcervical cancer were marked by Cy-3 and the samples of normal weremarked by Cy-5.(4)hybridization:The mixed probes were hybridized tothe cDNA microarray H20K2L(16000 gene points). Each sample wasrepeated 3 times.(5)checking and analysis: After high –stringentwashing ,the cDNA microarry was scanned for the fluorescent signals bythe scanner AFFYMETRIX428 ARRAY , then the fluorescent signals weretreated with ImaGene4.2 to change into figures to show differentiallyexpressed genes between each 2 tissues. All the figures were revised withhousekeeping gene. picking up the genes which expressed diferentlly in allthe 9 microarrys ,and the genes were the results which based the standardthat the difference between two figures was over double . Result:By applying this chip assay to the cancerous tissues and thepara-cancerous normal tissues, we found that Gene 616,567 and 522expressed differentially between cancers and its corresponding normaltissues. The downregulated genes is more than the upregulated genes,including the genes about cell apoptosis ,anti-oncogene, signal transmission,ribosomal protein, protein composition and extending, et al. theupregulated genes included the genes about oncogne,cell anti-apoptosisgenes, tumour metastasis, metabolism, immunity, protein composition, et al.We identified 12 differentially expressed genes in all the 3 cancersamples,and we further identified 3 upregulated and 9 downregulated genesfor functional analysis. The upregulated genes included MMP-12,ILE1 andhTERT. The downregulated genes included 4(AQP4),RPL23A,NFKappaB,nm23,CALU,P-120,FHIT,UBL3 and LOC255147 . Conclusions:(1) Cervical squamous cell carcinoma is the result towhich balance of many genes expression were broken.the genes includeMMP-12,ILE1 and hTERT . The downregulated genes included 4(AQP4),RPL23A,NF KappaB,nm23,CALU,FHIT,P-120,UBL3 andLOC255147 . (2) cDNA microarray is a powerful strategy to indentifynovel cancer associated genes, and it can rapidly explore the roles of thenovel genes in clinical cancer rapidly and efficiently. we can investigate thegenes in big flux with cDNA microarray .