Relation Between HPV16/18 And HCCR In Cervical Intraepithelial Neoplasia And Cervical Cancer

IntroductionCervical cancer (CC) is one of the most frequently founded malignant tumors in women world wide. Current evidences implicated high risk humn papillomavirus (HPV) especially HPV16/18 infection as a causative part of the natural history of cervical carcinogenesis. HPV can be found in 99.7% of cervical cancer patients. It's critical for precaution of cervical cancer to detect HPV. However only a part of the patients who contracted HPV may advance to cervical cancer. Cervical cancer oncogene (HCCR-1, HCCR-2) was overexpressed in cervical cancer and cervical intraepithelial neoplasia (CINI, II, III) compared with it in normal cervical tissue. So detection of HCCR protein and HPV16/18 in cervical exfoliated cells or tissue is very important for diagnosis at cervical cancer and precancerous lesion. In our study, we detect HPV16/18 by the hybridization in situ and HCCR by immuohistochemical analysis, to research the value of them in cutting down the mortality of CC, and to supply rationale to exploitation of convenient samples.Materials and Methods1. Materials(1) specimen570 cervical liquid-based cells remained samples were obtained from the Department of gynecology in the First Affiliated Hospital of China Medical University, during Nov 2005 to Nov 2006. All of them were treated as CC or CIN, Among them 31 samples were used in our study, in addition 4 samples of normal and 6 samples of inflammation patients as control gropes. All of them possess corresponding histological samples as controls. The patients with CC were 6, with CINI were 11, with CINII\III were 14. The average age is 46.7(range, 25～68 years). The standard of cytology grouping based on histodiagnosis. All patients had not the history of the operation on cervix, pregnancy and radiation therapy in the cavity of pelvic.(2) Main agents, equipment and sourceKit of HPV16/18 in situ hybridization was purchased from MAIXIN biological technology company in Fuzhou. antibody of HCCR protein was friendly presented by Jin woo Kim professor in Korea. S-P Kit and DAB Kit were obtained from MAIXIN biological technology company in Fuzhou.2. Methods(1) Detection of HCCR by Immuohistochemical analysis S-P methods. Result determination: The overexpressed staining was in the cytoplasm. The number of staining cells were graded as follows: 10-24%(+); 25-49%(++); 50-74%(+++);≥75%(++++).(2) Detection of HPV16/18 by Hybridization in situ. The masculine staining was in the nucleus.(3) Statistical analysisAll the datas were analysis with x~2 test and Fisher's test, P=0.05. The relation between HPV16/1 8 and HCCR was analysis with spearman analyses, P=0.05.Results1. Expressions of HCCR in cervical exfoliated cells and cervical tissue by Immuohistochemical analysis S-P methods. HCCR was overexpressed in cervical cancer(100%) and CINI(72.7%), CINII/III(78.6%) compared with that in normal cervical exfoliated cells and tissues (0%), p < 0.005. The expression of HCCR in cervical cancer and precancerous lesion correlated well with that in cervical neoplasia (CIN). It's consistent both in cervical exfoliated cell and in cervical tissue.2. Expression of HPV16/18 by Hybridization in situ. HPV16/18 was overexpressed in cervical cancer(83.3%) and CINII/III (71.4%) compared with it in normal(0%) and inflammation cervical exfoliated cells and tissues (50)%), P < 0.005. The expression of HPV16/18 in cervical cancer and precancerous lesion correlated well with cervical neoplasia (CIN). It's consistent both in cervical exfoliated cell and in cervical tissue.3. Overexpression of HPV1/18 and HCCR showed a significantly positive correlation, r_s=1. P < 0.005.DicussionCancer of cervix is a principle jeopardizing female lives. HR-HPV 16/18 is its positive pathogenetic cause, and therefore it's extremely necessary for developing the screening between HPV infection and cervix affection.Human cervical cancer oncogene (HCCR) is a new oncogene which was found in 2003. HCCR don't express in normal cervical tissue, but overexpressed in cervical cancer. HCCR inhibit P53 by binding with it to cause the genesis and development of cervical cancer.Not all patients who infected HPV will proceed to cervical cancer, it's not very accurate to screen CC only by detection of HPV. But detection of HCCR and HPV16/18 at the same time will be much worthwhile.Our outcome shows that with the increase of serious grade in CIN and CC, positive expression rates of HPV/16/18 and HCCR increase too. Positive expression rate of HPV16/18 in inflammation is lightly lower than that in CINI, but that of HCCR is obviously lower than that in CINI. Therefore, detection of HCCR in patients who were positive in HPV16/18 may predict the evolution and development of CIN. It can direct the patients to make biopsy properly so to decrease the rate of missed diagnosis of CIN and CC and to guide doctors dealing correctly with positive patients of HPV16/18.Our research showed that the detection results of HPV16/18 and HCCR in remained samples of liquid-base cells and cervical tissues are coordinate, which verified that remained samples of liquid-based cells can be used to study of molecular biology. Therefore, we can simplify the procedure and reduce the cost of obtaining samples.Conclusions1. In CIN and CC, The positive rates of HCCR and HPV16/18 increase with the aggravation of cervix affection.2. In CIN and CC. the expression rate of HCCR and HPY16/18 showed a positive correlation, It will be more accurate for screening CIN and CC to detect them together.3. The positive rate of ether HCCR or HPV16/18 in the remained samples of cervical liquid-based cells is coincident with that in cervical tissues. There is great feasibility in screening cervical affection by utilization of the remained samples of cervical liquid-based cells.