The Study Of P27～(kip1) And Skp2 Protein Express In Between Cin And Cervical Carcinoma And Clinical Signifcance

Objective:To detect the expression level of cyclin-dependent kinase inhibitor, P27Kip1 and S-phase kinase associated protein 2, Skp2 in cervical intraepithelial neoplasia (CIN) and Squamous cell Carcinoma of Cervix (SCC) . Skp2 mRNA expression was examined by semiquantitative PCR in fresh cervical carcinoma tissue and normal cervical tissue.And to discuss the biological and clinical significance of Skp2 and P27Kip1 experssion in CIN and SCC and the relationship between the expression of Skp2 and P27Kip1. Then try to analysis the relationship between dysregulation of the cell cycle in the occurrence and development of cervical tumors.Methods:Immunohistochemistry was performed in the 10 normal cervical tissue and 43 CIN and 34 SCC to evaluate the alteration of the P27Kip1 , Skp2 proteins. RT-PCR was performed in fresh normal cervical tissue and SCC to detect the expression of Skp2 gene and to analysis the relationship between Skp2 and P27Kip1 expression and clinicopathological characteristics. Result:1 The positive expression rate of P27Kip1 protein in normal cervical epithelium, CIN and SCC was 90%(9/10), 48. 84%(21/43), 29. 41%(10/34)respectively. We found that the expression of P27Kip1 in these three lesions existed statistics difference(P<0.05).2. The positive expression rate of Skp2 protein in normal cervical epithelium , CIN and SCC was 0(0/10), 39. 53%(17/43) and 70. 58%(24/34) respectively.We found statistics difference existed in these three lesions of expression of Skp2(P<0. 05). RT-PCR analysis showed that Skp2 mRNA was overexpressed in SCC(1. 2771 ± 0. 2741) compared with normal cervical epithliume(0. 6380±0. 1840), Statistical difference was found between them (P<0. 05).3. The positive rates of P27Kip1 in CIN Ⅰ — Ⅲ were 53.84%(7/13), 66. 67%(8/12), 33. 33%(6/18) respectively,The positive rates of Skp2 in CIN Ⅰ ~ Ⅲ were 23.07%(3/13), 33.33%(4/12), 55.56%(10/18) respectively ;No statistical difference of Skp2 and P27Kip1 proetins expression was found between CIN Ⅰ —Ⅲ.4. The positive rates of P27Kip1 in high, medium, low differentiation SCC were 36. 36%(4/11), 26. 67%(4/15), 25. 00%(2/8) respectively. The positive rates of Skp2 in high, medium, low differentiation SCC were 54. 54%(6/11), 80.00%(12/15), 75.00% (6/8)respectively; No statistical difference of Skp2 and P27Kip1 proteins expression was found among SCC differentiation degree.5. The expression of P27Kip1 29. 41%(10/34) in SCC was correlate with the expression of Skp2 70. 58%(24/34) in SCC and it was negative correlation(P <0. 05).6. Correlation was existed between the expression of P27Kip1, Skp2 and lymph node metastasis. The positive rates of P27Kip1 and Skp2 were 68. 18%(15/22) , 36.36%(8/22) in SCC without lymph node metastasis and were 33.33%(4/12), 83.33%(10/12) in cases with lymph node metastasis. The positive rates of P27Kip1, Skp2 in SCC with lymph node metastasis wrer significantly lower/higher than those without lymph node metastasis(P<0. 05). No significant relationships were found between the expression of P27Kip1 , Skp2 and other clinicopathological features, such as age, tumor type, clinic grade, clinic stage, etc.7. Kaplan-Meier survive curve analysis showed survival rate of P27Kip1 positive expression was higher than that of P27Kip1 negtive expression. By Log-Rank test Statistical difference was found between them (P<0. 05).8. Kaplan-Meier survive curve analysis showed survival rate of Skp2 positive expression was lower than that of Skp2 negtive expression. By Log-Rank test Statistical difference was found between them (P<0. 01).9. In COX proportional hazard regression model analysis, Skp2 overexpression and lymph node metastasis were identified as significant prognostic parameters and were two independent prognostic marker of SCC patients. Relative risk of the positive expression of Skp2 was 3.335,x2 was 4. 702 and P<0. 05;Relative risk of lymph node metastasis was 4. 936,x2 was 7.481 and p<0.01.Conclusions:1. The expression of P27Kip1 in normal cervical epithelium, CIN and SCC were all showed significant difference, The expression of P27Kip1 tends to decrease with the development of tumor in different cervical diseases. From the research it suggested that the decrease of P27Kip1 expression may play a important role in cervical epithelium malignant transformation .2. The expression of Skp2 protein in normal cervical epithelium, CIN and SCC were all showed significant difference, The expression of Skp2 tends to increase with the development of tumor in different cervical diseases. Skp2 expression might play an important role in the development and progression of SCC. Skp2 mRNA was up-regulated in SCC, the positive expression rate of Skp2 protein was raised in carcinoma tissue. In the present study it suggested overexpression of Skp2 had a stonger invasionand poor prognosis in SCC patient. Moreover Skp2 overexpression is an independent prognostic marker of SCC.3. The level of P27Kip1 was inversely related to that of Skp2 in SCC.4. The result in this study suggested that Skp2, P27Kip1 be all involved in the carcinogenesis of SCC. overexpression of Skp2 modulate the malignant phenotype of cervical carcinoma possibly via P27Kip1 proteolysis. By this pathway P27Kip1 loss its tumor-suppression function, lead to cell cycle deregulation and cell malignant transformation then tumor formation.