The Association Study Of Abnormal Expression Of Skp2 And P27kip1 With Carcinogenesis And Clinicopathologic Characteristic Of Breast Cancer

Objective The relationship of abnormal expression of mRNA and protein of Skp2 and p27kipl in intraductal proliferation lesions was explored with carcinogenesis of breast cancer. The expression level of Skp2 protein in invasive carcinomas was detected in order to explore the association of abnormal expression of protein of Skp2 with clinicopathologic characteristics of invasive breast carcinoma. Method 1st Immunohistochemical method: The positive expression ratio of Skp2 protein and p27kipl protein in 120 cases of intraductal proliferation lesions including 30 cases of usual ductal hyperplasia, 30 cases flat epithelial atypia, 30 cases of atypical ductal hyperplasia and 30 cases of ductal carcinoma in situ were detected using immunohistochemical method .The expression of Skp2 protein in 56 cases of invasive carcinoma were detected and the relationship of the positive expression ratio of Skp2 protein were analyzed with metastasis of lymph nodes and histological grade . 2^nd Qualitative RT-PCR: Total RNA was extracted out of 60 cases of fresh tissue of intraductal proliferation lesions including 15 cases of UDH, 15 cases of ADH, 15 cases of FEA, 15 cases of DCIS and Skp2 mRNA and p27kipl mRNA were reversely transcribed into cDNA. cDNA were amplified by polymerase chain reaction . Product length: Skp2: 108bp , p27kipl:177bp . 3^rd Real time fluorescence quantitative RT-PCR: cDNA were made by reverse transcription reaction. The expression account of mRNA in intraductal proliferation lesions were detected by real time fluorescence quantitative RT-PCR. Product length: Skp2,108bp;p27kipl, 177bp. Results 1^st The positive expression ratio of Skp2 protein of DCIS, FEA andADH was significantly higher than that of UDH. (p=0.0O0, p=0.038, p=0.038) . There were remarkable differences between DCIS and ADH (p=0.030) ,DCIS andFEA(p=0.030) . No significan difference existed between ADH and \JDH(p=1.000). Significant difference of positive expression ratio of p27kipl protein existed between UDH. and DCIS (p=0.000), FEAand DCIS (p=0.010), ADH and DCIS(p=0.039).No significan difference existed between ADH and FEA(p=0.795), ADH and UDH(p=0.069), UDH and FEA(p= 0.192). Skp2 protein level was reversely correlated with p27kipl protein level in intraductal proliferation lesions(r=-0.411,p=0.000). The same results existed in UDH and DCIS(r=-0.406,p=0.026;r=-0.544,p=0.002) . 2nd The positive expression ratio of Skp2 protein in invasivecarrcinoma accompanied with positive lymph nodes were remarkably higher than negative lymph nodes (p<0.025). Difference of positive rate of Skp2 exists in histological grade Ito2 and grade 3 of invasive carcinoma (p<0.01). 3rd Skp2 mRNA and p27kipl mRNA were detected in intraductal proliferation lesions. Product length: 177bp. The 98 % of sequencing result of product of PCR accorded with the sequence of gene bank. At the same time, the amount of Skp2 mRNA in DCIS was more than that in UDH(p=0.000), FEA (p=0.000)and ADH (p=0.000). There was great difference between FEA and ADH (p=0.010)as well as between FEA and UDH(p=0.004) . No significance difference existed between ADH and UDH (p=0.750). However, reverse results of amount of p27kipl mRNA existed infour groups, such as between DCIS and FEA (p=0.000) , between DCIS and ADH 0?=0.000), DCIS and UDH (p=0.000), between FEAand ADH (p=0.000). There was no significance between ADH and UDH (/?=0.137) . There was reverse association between Skp2 and p27kipl in intraductal proliferation lesions(r=-0.741, p=0.000). Conclusions 1st The increase of expression level of Skp2 protein anddecrease of expression level of p27kipl protein have closed association with dysplasia of breast ductal epithelial and carcinogenesis of breast cancer. 2nd The invasive carcinoma owing to higher level of Skp2 protein with stronger ability of extension to lymph nodes and higher histological grade worse prognosis have worse prognosis. 3rd It was supposed that abbreviation amount of Skp2 mRNA and p27kipl mRNA has effect on dysplasia of ductal epithelial of breast and carcinogenesis of breast cancer.