The Study Of Relationship Between Expression Of P27～(kipl), CyclinE And Clinicopathological Parameters And Prognosis Of Colon Cancer

PurposeLike other tumors, the occurrence of colon cancer is induced by the deregulation of cell cycle resulting in the abnormal proliferation and differentiation of cells. The running of cell cycle is regulated by cycling CDK and CDKI in many level. The conjugation of the former two can promote the progress of cell cycle. CDKI is a negative regulative factor of cell cycle, and it can block progression of cell cycle, inhibit over - proliferation of cells and participate proliferation, differen-ciation and apoptosis of tumor cells by inhibiting formation and biological activity of the cyclin - CDK complex. P27 is amultifunctional CDKI. It is showed that p27 kip1 and cyclinE had abnormal expression in many kinds of malignancies. Such as stomach, breast, lung, prostate, head and neck cancer and leukemia, but there is no report on the expression of them and the correlation between them in colon cancer in our country. In our experiment, we used immunohistochemistry to study the expression of p27kip1 and cyclinE in colon cancer and explore the relationship between their biological between their biological behaviour and prognosis.Materials一. Sample:Tumor experimental group: there were 71 paratfin embeded species of colon cancer which came from Large Intestinal Surgery of Lia-oning Tumor Hospital between 1994 to 1995. Among tern, 33 cases were male and 38 were female. Their age range was from 42 to 79 ( mean 65 years old). Tumor clinical pathological staging was according to China Modified Dukes Staging Standard. 35 DukesA, B, 20 DukesC and 16 Dukes D stage. Tumor cell differentiation was divided into high, moderate, low and undifferentiated degree and mucoid gland in cancer and King's cell cancer were classified into undifferentiated group. In our experiment, 33 high, 19 moderat and low and 19 undifferentiated degree.Normal contrast group: there were 21 cases colon mucosa species taken at 10cm away from the resected section.二. Main Reagent:Mouse anti - human monoclonal antibody against p27kip1 and cy-elinE respectively, SP reagent box and DAB visualizing agent.Experimental MethodSP immunohistochemical staining; paraffin wax slides were deparaffinized by bi - methyl - bezidine and hydrated by alcohol; antigen repaired by high femperature and high pressure; adding 3% H2O2 - for 10 minutes at room femperature; PBS washing for 3 times, 5 minutes each time j adding 10% normal goat serum and incubatingat 37 ℃ for 20 minutes; PBS washing 3 times, five minutes each time; adding biotin - labeled secondary antibody worlcing solution and incubating for 30 minutes at 371 ; PBS washing three times, five minutes each time; incubating for 30 minutes at 31 ℃. Strep lividin washing 3 times with PBS, 5 minutes each time; DAB visualizating; washing with water; counter staining slides with Harris'hematoxytlin completely and coverslipping.Result1. Among 71 cases of colon cancer species, 35 cases were positive expression of p27kip1 positive expression rat was descendent with ascending of Dukes stage. There was a significant difference between Dukes stage and Dukes C or D stage. ( p =0. 028,0. 022). But there was no difference between Dukes C and D stage ( p = 0. 722). In plasma infiltrated group. The expression rate of p27kip1 was decreased and there was a significant difference comparing with that of plasma un-infiltrated group (p =0.002). Moreover, the positive expression rate of p27kip1 poor was decreased in lymph node metastases group and there was a significant difference comparing with that of non -metastares group ( p = 0. 042). The p27 expression rate was decreased gradually following the sequence of high, moderate, low and undifferentiated group, and there was a significant difference between them respectively ( p = 0. 015,0. 001,0. 031) There was no correlation between p27 expression and grossly type or proportion of tumor and colon diameter.2. Among 71 cases of colon cancer species, there were 33 cases of positive cyclin stained (46.5% ). Among cyclinE positive expres...