| Objective1）To study the infection state of HR-HPV in cervical cancer tissues and itsrelationship between cervical cancer occurrence and HPV infection.2）To screen miRNAexpressed profile of HPV-positive cervical cancer and normal cervical epithelium, tomodulate the special miRNA of HPV-positive cervical and investigate its effects onbiological behavior of HPV-positive cervical cancer,and to investigate the gene expressedchanges in HPV-positive cervical cancer．3）To study the biological function of miRNA invitro experiment of HPV-positive cervical cancer cells.Methods1）HR-HPV infection was detected by HC2,30cases of HR-HPV positive cervicalcancer tissues were collected, the expression of HPV16-DNA was detected by using thenested PCR, existence state in the organization of HPV16-DNA was detected by themultiplex real-time quantitative PCR. analysis2）The miRNA expressions in6pairs ofHPV-positive cervical cancer and normal cervical tissues were measured by miRNAmicroarray,and the real time PCR was used to validate the microarray results；Differentially expressed miRNAs were then identified through fold change as well as Pvalue calculated using t-test. The threshold set for up-and down-regulated genes was a foldchange>=2.0and a P value﹤0.05. Target genes of differentially expressed miRNAs werethe intersection predicted with3database（sTargetscan，microRNAorg，PITA）. HierarchicalClustering was performed to show the distinguishable miRNAs expression pattern amongsamples.3)According to the microarray results,we select19of differentially expressedmiRNAs and5of differentially expressed target genes, and the real time PCR was used tovalidate the microarray results,and we make statistical analysis by combined with clinical pathological data;4) We screened out150f the down-regulated miRNA in SiHa cells,which were packaged by Lentivirus,and screened out some fuctional miRNA by the HCS.Results1)HPV-16positive rate in patients with cervical squamous cell carcinomas (SCC)was96.7%,Compared with HC2detection rate no significant correlation were found (P﹥0.05); the HPV-16DNA exists in integrate state in cervical cancer tissues, its integraterate was89.7%. The expression level of HPV-16DNA has nothing to do with age andlymph node metastasis of cervical cancer patients,but it is significantly associated withpathological classification and clinical staging of cervical cancer patients;2）It identified140differentially expressed miRNAs betweent HPV-positive cervical cancer and normalcervical epithelium by the miRNA microarray,and among them640f them wereup-regulated miRNA,76of them were down-regulated miRNA. Real-time PCR validatedthat a part of differential expression of miRNAs is equal in result with the miRNAmicroarray,but a part of result were reverse. Coincidence rate of gene chip is73.8%.3)Relative expression of19miRNAs was significantly different in cancer tissues thanin normal tissues(P﹤0.05); The expression level of miR-96, miR-196, miR-17, miR-21,miR-15was significantly associated with pathological classification, invasive depth andclinical staging of cervical cancer (P﹤0.01),but them have nothing to do with age,tumorsize and menopause (P﹥0.05);4) The miRNA-1224, miRNA-Let7c significiantlyinhibited the profiferation of tumor cells in vitro.Conclusion1)HR-HPV infection is the main factor lead to cervical cancer, and HPV-16is main types of viruses in xinjiang uygur cervical cancer, its integration status in theorganization plays a main role in the incidence of cervical cancer and is closely related tocervical cell malignant transformation.2)Aberrant expression of miRNA-196,miRNA-17,miRNA-15,miRNA-21suggest they might play a important role as an oncogene in thetumorigenesis and development of cervical carcinoma and it possibly correlates with progression and prognosis of cervical cancer.,3)The miRNA-1224, miRNA-Let7c significiantlyinhibited the profiferation of tumor cells in vitro. |
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