The Expression And Clinical Significance Of HPV16-E6,PCNA And INOs In The Process Of Cancerating Of Cervical Epithelium

Objective:. Cervical cancer is common gynecologic tu-mors and in the world it is the second most common canc-erin women after breast cancer, but its mortality rate in w-omen is the highest. Large epidemiological and biological research shows that persistent infection caused by high-riskHPV is the major risk factor for cervical cancer and prec-ancerous lesions. Thus,the identification of the type of HPV,particularly high-risk subtypes,has great significance to the early detection and prevention of cervical cancer. Proliferati-ng cell nuclear antigen is a cell cycle regulatory protein f-or DNA synthesis, and it is also a common indicatorfordet-ecting tumor cell proliferation. Inducible Nitricoxide syntha-se is a non-calcium-dependent enzyme. It can induced by LPS and inflammatory mediators and produce nitric-oxide.To discuss the effects of HPV-E6,PCNA and iNOs on the genesis,progression and metastasis of carcinoma of the ute-rine cervix,we use immunohistochemical methods and flowcytometry method to detect the expressions of HPV16-E6 p-rotein, PCNA, iNOs in normal cervical tissue, Chronic cer-vicitis,CIN and cervical cancer and the relati-onship betwe- en them.Methods:All the specimens were recruited from the p-atients with cervical tissues who were treated first in the gynecology department of the Fourth Affiliated Hospital of Hebei Medical University from January 2006 to June 2007.All the cervical cancer were confirmed by pathologic obse-rvation. 52 cervical carcimona, 40 cervical intraepithelial n-eoplasia, 9 Chronic cervicitis and 3 normal cervical tissueswere recruited.Immunohistochemical technique and flow cyt-ometry were performed to examine the expression of HPV16-E6 protein,PCNA and iNOs of the cervical affection t-issue. Analyze the relationship between the expression andt-he clinical pathological characteristics, and the correlation between the expression of HPV16-E6,PCNAand iNOs protein in the cervical affection tissue through the statistical an-alysis with the results above by SPSS13.0statistic sotware.Statistical method concludes Chi-square test, Fisher exactprobabilitics, partitions of x2method Spearman rank correlationshomogeneity of variance and LSD test.Results: 1 The relationship between the HPV-E6 expression and clinical pathological features:The positive rate of HPV-E6 protein was 90.38,60.00%,25.58%,0% in cervical cancer, CIN, chronic cervicitis and normal cervical tissues respectively. It decreased and there is an obvious distinction among four group. The positive rate of HPV-E6 is 92.59% in the patients of cervical cancer whose age is less than 45, by contrast, the positive rate of HPV-E6 is 88.00% whose age is not less than 45. The difference between them had no statistics meaning. The positive rate of HPV-E6 is 97.50% in the squamous cancer, by contrast, the positive rate of HPV-E6 is 66.67% in non-squamous cancer. There is obvious distinction between them. HPV-E6 can be expressed differently with different histological grade. GradeⅠ～Ⅱis 84.85%. GradeⅢis 89.47%. The difference among them had no statistics meaning. HPV-E6 can be expressed differently with different FIGO stage. stageⅠis 88.00%. stageⅡ~Ⅲis 89.47% and the difference among them had no statistics meaning. The positive rate of HPV-E6 is 90.00% with lymphatic node metabasis, by contrast, the positive rate HPV-E6 is 90.63% without lymphatic node metabasis. There is no obvious distinction between them. The positive rate of HPV-E6 is 91.18% in the cervical tumorous focus whose diameter is less than 4cm, by contrast, the positive rate of HPV-E6 is 88.89% whose diameter is not less than 4cm. There is no obvious distinction between them;The positive rate of HPV-E6 is 95.00% with infiltration of latero-uterus or haemal tube, by contrast, the positive rate of HPV-E6 is 87.50% without infiltration of latero-uterus or haemal tube. There is no obvious distinction between them. The positive rate of HPV-E6 is 87.10% in shallow- muscular laye infiltration,by contrast, the positive rate of HPV-E6 is 95.24% in deep- muscular laye infiltration. There is no obvious distinction between them. 2 The relationship between the PCNA expression and clinical pathological features:The positive rate of PCNA protein was 86.54,56.00%,23.26%,0% in cervical cancer, CIN, chronic cervicitis and normal cervical tissues respectively. It decreased and there is an obvious distinction among four group. The positive r-ate of PCNA is 85.19% in the patients of cervical cancer whose age is less than 45, by contrast, the positive rate ofPCNA is 88.00% whose age is not less than 45. The diff-erence between them had no statistics meaning. The positi-ve rate of PCNA is 90.00% in the squamous cancer, and 77.78% in the adenocarcinoma of the uterine cervix,and the-n 66.67% in the other pathological type. There is no obvi-ous distinction between them. PCNA can be expressed diff-erently with different histological grade. GradeⅠ～Ⅱis 78.79%. GradeⅢis 100.0%.There is an obvious distinction among them. PCNA can be expressed differently with diff-erent FIGO stage.stageⅠis 72.00%.stageⅡ~Ⅲis 100.0% and had significant difference.The positive rate ofPCNA is 100.0% with lymphatic node metabasis , by contrast,the p-ositive rate PCNA is 78.13% without lymphatic node meta-basis. There is an obvious distinction between them. The p-ositive rate of PCNA is 85.29% in the cervical tumorous focus whose diameter is less than 4cm, by contrast, the po-sitive rate of PCNA is 88.89% whose diameter is not lessthan 4cm. There is no obvious distinction between them;T -he positive rate of PCNA is 100.0% with infiltration of la-terouterus or haemal tube, by contrast, the positive rate of PCNA is 78.13% without infiltration of latero-uterus or ha-emal tube.There is an obvious distinction between them.Th-e positive rate of PCNA is 77.42% in shallow-m-uscular l-ayeinfiltration, by contrast, the positive rate of PCNA is 100.0% in deep-muscular laye infiltration.There is an obviou-s distinction between them.3 The relationship between the iNOs expression and clinical pathological features:The positive rate of iNOs protein was 88.46,60.00%,27.91%,0% in cervical cancer, CIN, chronic cervicitis and normal cervical tissues respectively. It decreased and there is an obvious distinction among four group.The positive ra-te of iNOs is 88.89% in the patients of cervical cancer w-hose age is less than 45, by contrast, the positive rate of iNOs is 88.00% whose age is not less than 45.The differ-ence between them had no statistics meaning.The positive r-ate of iNOs is 90.00% in the squamous cancer, and 77.78% in the adenocarcinoma of the uterine cervix,and then 100% in the other pathological type.There is no obvious dist-inction between them. iNOs can be expressed differently with different histological grade. GradeⅠ～Ⅱis 87.88%. GradeⅢis 89.47%. There is no obvious distinction amon-g them. iNOs can be expressed differently with different FIGO stage. stageⅠis 76.00%. stageⅡ~Ⅲis 100.0% and had significant difference.The positive rate of iNOs is 100.0% with lymphatic node metabasis,by contrast,the positive r-ate iNOs is 81.82% without lymphatic node metabasis.There is an obvious distinction between them. The positive rat-e of iNOs is 85.29% in the cervical tumorous focus whos-e di-ameter is less than 4cm, by contrast, the positive rateof iNOsis 94.44% whose diameter is not less than 4cm. T-here is no obvious distinction between themThe positive rat-e of iNOs is 100.0% with infiltration of laterouterus or ha-emal tube, by contrast, the positive rate of iNOs is 81.82% without infiltration of latero-uterus or haemal tu-be. The-re is anobvious distinction between them. The positive rateof iNOs is 80.65% in shallow-muscular laye infiltration,bycontrast, the positive rate of iNOs is 100.0% in deepmuscu-lar laye infiltration. There is an obvious dis-tinction betwe-enthem.4 The relationship among HPV-E6,PCNA and iNOs: the relationship between HPV-E6 protein and PCNA was positive correlation in cervicitis and cervical cancer ,and HPV-E6 protein and iNOs was positive correlation ,and PCNA and iNOs protein was positive correlation.5 Flow cytometry results:the FI values of HPV-E6 protein in normal cervical tissues ,chronic cervicitis ,CIN and cervical cancer is 0.97±0.03,1.11±0.06,1.29±0.04,1.52±0.06 and there is an obvious distinction among four group. the FI values of PCNA protein in normal cervical tissues chronic cervicitis ,CIN and cervical cancer is 0.95±0.04,1.06±0.04,1.27±0.05,1.55±0.04 and there is an obvious distinction among four group. the FI values of iNOs protein in normal cervical tissues,chronic cervicitis ,CIN and cervical cancer is 0.96±0.04,1.10±0.06,1.42±0.08,1.84±0.09 and there is an obvious distinction among four group.Conclusions: 1 The positive rates of HPV-E6 protein innormal cervix, cervicitis, CIN and cervical cancer increase-ed gradually, and every two had an statistics meaning. Itshows that HPV16-E6 is were closely associated with gene-sis development and an HPV infection is an initial circum-stance in the genesis of cervical cancer. The expression ofHPV16-E6 is not associated with onset age,,degree of hist-odifferentiation, FIGO stage, tumour diameter, lymphatic n-ode metabasis, linfiltration of laterouterus or haemal tubea-nd infiltrative depth. while it is associated with pathologyi-cal type.2 The positive rate of PCNA in cervical cancer and CIN was significantly higher than cervisitis and normal cer-vical tissue, and the difference had an statistics meaning. which suggests that PCNA may cause cervical squamous c-ell hyperplasia and malignant transformation.The abnormal PCNA expression is an early event in cervical cancer. Theexpression of PCNA is not associated with onset age, path-ological type, tumour diameter, while it is assoc-iated with linfiltration of laterouterus or haemal tube, degree of histo -differentiation, FIGO stage, lymphatic node metabasis, and infiltrative depth.The PCNA expression may be related to the progress of the cervical cancer.3 The high expression of INOs can be considered that iNOS play an important role in the genesis and development in cervical cancer. The results showed that the expression of INOs is not associated with onset age, Organization size, type, histologic grade, while it is associated with Clinical stage, lymphatic node metabasis, infiltration of latero-uterus or haemal-tube and infiltrative depth. It suggests that INOS is closely related to the invasive cervical cancer.4 Cervicitis in the group, CIN and cervical cancer group HPV-E6 protein, PCNA and iNOs, every two of them are positive correlation.it Confirmed the HPV-E6 protein, PCNA, iNOs are closely related with the occurrence and development of cervical cancer, and they produce a synergistic effect to each other.