Expression Of MiR - 21 In Serum Of Patients With Cervical Squamous Cell Carcinoma And Its Effect On The Biological Characteristics Of Cervical Cancer SiHa Cells

PART1:EXPRESSION OF MIR-21IN SQUAMOUS CELL CARCINOMA OF CERVIX WITH LYMPH NODE METASTASIS AND ITS CLINICAL SIGNIFICANCEObjectiveLymph node metastasis is recognized as one of the major risk factors for cervical cancer prognosis cervical cancer, survival rate of patients with lymph node metastasis is far lower than that without lymph node metastasis.Early c-ervical cancer occur lymph node metastasis, the same staging patients whether there is lymph node metastasis, the prognosis is very different, and whether t-here is lymph node metastasis is closely related to the choice of treatment and surgery and postoperative radiotherapy.The study found that the occurrence of cervical cancer is closely related to the changes in miRNA. The majority of cervical cancer pathology is squamous cell carcinoma, In this study, Real-time RT-PCR detection the level of expression of serum microRNA-21(miR-21) in cervical squamous cell carcinoma with lymph node metastasis, no lymph node metastasis of cervical squamous cell carcinoma and Cervical Intraepithelial Neoplasia, and investigate the expression and cervical squamous cell carcinoma lymph node metastasiswhether the relationship.To examine the level of expression of serum microRNA-21(miR-21) in cerval cancer patients and assess the diagnose function of serum miR-21in lymph node metastasis cases.MethodsA total of:80cases of cervical cancer at the Guangxi cancer hospital, Dep-artment of Gynecologic Oncology, between2011January and September were selected, among the tatal,25cases were cervical cancer with lymph node meta-stasis (LNM),40cases were cervical cancer without LNM,15cases were normal control. Examine the level of expression of serum miR-21by quantitative reve-rse transcription polymerase chain reaction (qRT-PCR), analyzed by Pearson Correlation Analysis,draw the ROCcurve and find the best suitable cut-off point.Results1.The serum miR-21level of cervical cancer cases with LNM is higher than the cases without LNM and normal control, P<0.05.2.Serum miR-21level and LNM in cervical cancer were correlation with each other(Spearman Correlation Coefficient=0.408, P<0.001). Areas under the curve (AUC) was0.770.The sensitivities and specificities of serum miR-21were56%and92.5%, respectively, cut-off points for LNM cases was6.65.Conclusions Serum miR-21may be used to estimate LNM and could be a useful tool in making therapeutic decisions in cervical cancer. PART2:EFFECTS OF MIR-21ON BIOLOGICAL PROPERTIES OF SIHA CERVICAL SQUAMOUS CELL CARCINOMAObjectivecervical cancer is the mostly common tumor of reproductive system in female.Cervical cancer of main pathological types is cervical squamous cell carcinoma and cervical adenocarcinoma. High-risk human papillomavirus (human papilomavirus, HPV) plays an important role in the malignant transformation of cervical epithelial cells. HPV16is connected with cervical squamous cell carcinoma,and HPV18is connected with cervical adenocarcinoma. But the specific molecular mechanism of cervical cancer is still unclear. In this study, using RNA interference (RNAi) technology to change miR-21expression in siha cells derived from human cervical squamous cell carcinoma (HPV16positive), The study using a technique called RNA interference (RNAi) change miR-21expression in SiHa cells(HPV16positive)that comes from cervical squamous cell carcinoma of the people, to investigate the change of proliferation and migration invasion in SiHa. Explore the effects of miR-21on biological properties such as Proliferation、migration and invasion of SiHa. To provide the experimental basis for cervical squamous cell carcinoma of early diagnosis and gene therapy and prognosis. MethodsSelected SiHa cells come from cervical squamous cell carcinoma of the pe-ople, By using artificial synthesis of miR-21simulation objects miR-21-mimic and miR-21restrain objects miR-21-inhibitor. The SiHa cells were divided into three groups:miR-21-mimic group, miR-21-inhibitor group and blank control group. Through Lipofectamine TM2000,the SiHa cells were transfected with miR-21-mimic、miR-21-inhibitor and blank control, Through the flow cyto-meter(FCM) detection transfect rate, after SiHa cells were transfected48hours, The miR-21expression was measured by quantitative Real Time-PCR. Cellular proliferation capacity were assayed by the methyl thiazolyl tetrazolium (MTT) assay.Cellular migration and invasion ability were detected by Transwell cham berassay.Results:1.The result of quantitative Real Time-PCR showed:after incubation with miR-21-mimic for48h,the level of miR-21in miR-21up-regulation group was up-regulated significantly compared with that of the control group(P<0.01),while the level of miR-21in miR-21down-regulation group was decreased by anti-miR-21oligonucleotides compared with the control group(P<0.01).2.The result of MTT assay showed:after incubation for48h,the cell proliferation ability of the miR-21up-regulation group was increased compared with that of the control group(P<0.05);while the cell proliferation ability of the miR-21down-regulation group was reduced compared with that of thecontrol group(P<0.05).3.The result of Transwell invasion assay showed:compared with that of the control group,the number of cells in the miR-21up-regulation group which cross the wound region was more significantly after incubation for48h;on the contrary,the number of cells in miR-21down-regulation group which cross the wound region was less compared with the control group.4.The result of Transwell invasion assay showed:There were different significances between the miR-21up-regulation group and the control group(P <0.01)and there were different significances between the miR-21down-regulation group and the control group(P<0.01).Conclusions:1.miR-21-mimic can up-regulated the level of miR-21and anti-miR-21oligonucleotides can down-regulated the level of miR-21in SiHa cells.2.After the up-regulated expression of miR-21,the cellular proliferation ability was enhanced and the activity of cellular migration and invasion were positively regulated in SiHa cells;after the down-regulated expression of miR-21,the ability of cellular proliferation、migration and invasion were inhibited.3.miR-21is closely related with the development of cervical cancer.