The Significance Of The Study Of Mirna-155,hpv16/18by Self-Made Tissue Microarray In The Occurrence And Development Of The Cervical Cancer And The Clinical Pathological

Objective: Cervical cancer is one of the most common cancer.Therewere about466,000new cases every year all over the world and80%caseshappen in developing countries.As a result,the cervical cancer has alreadybecome the most common cancer and the main cause of death among womenin developing countries. Its death rate follows closely after the breast cancer,so it is the second leading disease of death among women, in recent yearsincidence is younger and constantly on the rise, which threaten the life andhealth of women seriously. Its occurrence and development are thepathological process involved multi-factor and multi-procedure.So far, itspathogenesis has not yet fully elucidated. In the past few decades, scholars athome and abroad has always kept studying the mechanism causing thecervical cancer. Numerous Studies of its pathogenesis from the protein level tothe gene level, which are of great significance for the clinical diagnosis,differential diagnosis, and guidance of clinical treatment.Many researches confirm that the principal factor of causing the cervicalcancer is due to the infection of human papillomavirus(HPV), in particular, thepersistent infection of high-risk HPV16/18relate closely to the cervicalintraepithelial neoplasia,(CIN),and the occurrence and development ofcervical cancel. The carcinogenic process of HPV16/18has oftenaccompanied with proto-oncogene mutation or antioncogene inactivation.Though the high-risk HPV infection is a necessary condition for cervicalcancer, a pure HPV infection is not sufficient to induce it. Therefore,otherunknown genes may be involved in this process, microRNA (miRNA)is a kindof endogenous non-coding small RNA composed of21-25nucleotides, widelyexists in animals and plants. It can involve in gene transcription regulation, joins the regulation of body’s various physiological processes, as well asregulate the gene’ expressive activity. In certain disease, the expression can beobviously up and down, which has attracted great notice from the scholars allover the world. The miRNA fragments of the third Exon on BIC gene.The miRNA-155is a small, typical multi-functional RNA and encoded by21-22nucleotides. A growing number of studies have shown that miRNA-155plays an important role as oncogenes in many tumors. Some studies haveaffirmed that miRNA-155can show abnormal expression in the process ofcervical cancer. But people have different ideas whether its expression isincreased or decreased.Due to the limitation of our technical level, there are many questions aboutwhat is the pathogenic mechanism of miRNA-155in the cervical cancer andwhat kind of connection does the miRNA-155have with the cervical lesions.The issue will detect the expression of miRNA-155and the infection ofHPV16/18in cervical cancer, CIN pathological changes, cervical squamouscell carcinoma through self-made tissue microarray and in situ hybridizationtechnique to observe the expression of miRNA-155in different lesions, tostudy the role of the virus in cervical carcinogenesis and discuss theconnections among miRNA-155HPV16/18and the different lesions ofcervical, and meanwhile, the likely relationships of the thereto analyze nowthe HPV16/18infection can affect the expression of miRNA-155so as to finda more objection can affect the expression of miRNA-155,so as to find a moreobjective and reliable indicators for clinical monitoring the progress of thelesions, as well as provide an evidence for further revealing the mechanism ofHPV carcinogenic and synergy factors. There is an important directivefunction for early preventions, early detections and early treatment of cervicalcancer.Methods:1Collect from HEBEI provincial people’ Hospital,200cases of cervical tissuesamples and surgical samples, of which31cases cervix uterus tissuematched group and different126cases CIN tissue research group (including 31cases of CIN Ⅰ,47cases of CIN Ⅱ,48cases of CIN Ⅲ),43cases ofcervical invasive squamous cell carcinoma tissue research group.2Apply the self-made tissue microarray; put the200cases of donor samples inarray design order into the recipient of paraffin block, then fusing, slicing,gaining, baking processes and so on, and then the tissue microarray is made.3Using in situ hybridization to examine the expression of miRNA-155incervical squamous cell carcinoma, various graded cervical CIN tissues andnormal cervical squamous epithelial tissue.4In situ hybridization (ISH) is used to examine the expression of HPV16/18in cervical squamous cell carcinoma, cervical CIN tissue and normal cervicalsquamous epithelial tissue.5Using the SPSS16.0software system for statistical analysis, data-countingneed x2test. Graded data use non-parametric rank sum test andcorrelation.Using the spearman rank analysis. Using a=o.005as the inspectionlevel which is statistically significant when p <0.05.Results:1The preparation of tissue chip and staining resultsAccording to the subject of design and the requirements of test method,3tissue chips are needed in8×8,8×9array. Dots of tissue chip lined up order,uniformed in size and no shifting,10tissue dots were absence and29dotsdon’t have significant tissue structures, result in a meaningful sample of162cases(36cases of cervical squamous cell carcinoma,29cases of CINⅢ,40cases of CIN Ⅱ,26cases of CIN Ⅰ,31cases of normal cervical squamousepithelium)the observable rate of the tissue dots’ morphology is81%.The HEstaining was uniformity and no dropping, moving and winkling, in situhybridization staining positive positioning is clear, the background is alsoclean.2Expression of HPV-DNA16118in different cervical intra epithelial lesions.Apply the in situ hybridization to examine the expression ofHPV-DNA16/18.In36cases of cervical squamous cell carcinma,95cases ofCIN (including29cases of CINⅢ,40cases of CINⅡ,26cases o CIN Ⅰ) and 31cases of normal cervical squamous epithelial tissue. The results showed:the HPV-DNA in the normal cervical squamous epithelium has no expression,from CIN to cervical squamous cell carcinoma, the expression of HPV-DNAwere50%(13/26),67.5%(27/40),79.3(23/29),52.8%(19/36)respectively,thedifference was statistically significant(p<0.05).The positive rate of HPV-DNA.In the graded CIN lesions and cervical squamous cell carcinoma wasobviously higher than that in the normal cervical squamous epithelium(p<0.05).But between the CIN grade and CIN grade, CIN and cervicalsquamous cell and carcinoma. The difference in the expression of HPV-DNAwas not statistically significant.(P>0.05).3Expression of mir-RNA155in different cervical intraepithelial lesionsApply the in situ hybridization to examine the expression of mir-RNA-155.In36cases of cervical squamous cell carcinma,95cases of CIN (including29cases of CIN Ⅲ,40cases of CINⅡ,26cases o CIN Ⅰ) and31cases of normalcervical squamous epithelial tissue. It shows the result: in normal Cervicalnon-keratinizing squamous epithelium miRNA-155maily expresses high incytoplasm64%(20/30), positive rate of miRNA-155in CIN Ⅰ, CINC, CINⅢand ISCC expression respectively65%(17/26),45％(18/40),21％(6/29),22％(8/36) with lower expression of normal Cervical non-keratinizing squamousepithelium and with significant difference (p<0.05) and weakened in turn. Butthere are no difference of positive rate of miRNA-155among CIN Ⅰand CINⅡ, CIN Ⅱ and CIN Ⅲ, ISCC and CINⅢ.(p＞.005)4Correlation of the expression of P16and HPV16/18infection in cervicallesions.There is no relativity of expressing of cervical squamous cancer HPV16/18and miRNA-155.(p＞0.005)5The relationship between expression of miRNA-155, HPV-DNA16/18andpatient’s age, tumor size, Gleason score, status of lymph node and pathologicstage: expression of HPV-DNA16/18has no obvious relation with the abovementioned.(p＞.005)Conclusion: 1In situ hybridization detection HPV16/18has higher SPE, Positivepredictive value and accuracy in identifying high grade CIN and low grade CIN also has a high clinical prediction value towards the development of CIN.2In situ hybridization detection miRNA-155expresses high among normalcervical tissue and expresses inconspicuous among high grade CIN and ISCC;there isn’t higher SPE, Positive predictive value and accuracy in identifyinghigh grade CIN and low grade CIN; it is trivial towards the development ofCIN and clinical prediction value.3There isn’t relativity of expressing of ISCC HPV-DNA16/18andmiRNA-155and no synergy in the ISCC diagnostic process of the two.4There is no synergy in expressing Expressing of miRNA-155、HPV-DNA16/18has no relativity with age, tumor size, Gleason score,lymph node metastasis and pathologic stage of the patient.5The procedures of handmade tissue microarray are simple, economical,convenient, so it is an easy and workable.