| Cyclin D1 plays an important role in cyclin family, it is seen as a growth factor receptors beca use it can promote cell cycle from G1 phase to S phase. It reaches the highest in G1 phase expressi on with periodic dependence, forming complexes through activates CDK4 and CDK6, positive reg ulation of cell cycle. Its abnormal can lead to cell cycle disorder and cell proliferation. P27 kip is one of the important members of KIP family which belongs to cell cycle insulin-dependent kinase inhibitors, it can inhibit the activity of a variety of cyclin/CDK complex, to negative cell cycle reg ulation. Ajind it also plays the role of mediating inhibitory cytokine reaction, promote differentiat ion and increase intercellular adhesion. P27 kip is independent prognostic factor in a wide variety o f tumor can be used as a sign of clinical diagnosis. In view of these two genes in tumor occurrence and inhibition of important for understanding the movement of breast tumor, to further understand cyclin D1 and p27 kip in gene and protein expression and mutual relations and the biological chara cteristics of these two kinds of protein and its application value in clinical diagnosis,we prepare do g cyclin D1 polyclonal antibody by prokaryotic expression technique and the lab has been p27 kip a ntibody, to make them in the CHMp, CHMm canine breast tumor cell line and to study the express ion of clinical specimens.The m RNA content of cyclin D1 in different cell lines was extremely significant differences(p<0.01), and extremely significant difference with normal mammary gland(p<0.01); the m RNA of p27 kip in the two cell lines were no significant difference, but the extremely significant difference with normal mammary gland(p<0.01); the m RNA levels of cyclin D1 and p27 kip were very significant differerce between each other in the same cell line(p<0.01). The antibody of cyclin D1 that we made by ourselves is very usefull, thought it express in inclusions, it has good reaction to the original, and high antibody titer(25, 600). The expression content of cyclin D1 and p27 was difference in differernt cell lines. The protein content of cyclin D1 in different cell lines was extremely significant differences(p<0.01), as well as extremely significant difference with normal mammary gland(p<0.01), and the protein expression of p27 kip in two kinds of cell lines have significant difference(p<0.01), and with the normal mammary gland extremely significant difference(p<0.01), the expression of cyclin D1 and p27 kip which in the same cell line is very significant difference(p<0.01). We have classified the clinical cases that we collected over the past five years, and got that 24 cases of benign tumors, 13 cases of intraductal carcinoma, infiltrating ductal carcinoma(19 cases), 10 cases of invasive lobular carcinoma, mucinous carcinoma in 5 cases, medullary carcinoma accompanied by lymphocyte infiltration(3 cases), and 10 cases of mixed carcinoma. In these cases, the positive expression rate of cyclin D1 were 58.33%, 40.00%, 78.95%, 90.00%, 60.00%, 100.0% and 90.00%; the positive expression rate of p27 kip were 62.50%, 23.08%, 42.11%, 50.00%, 60.00%, 33.33% and 70.00%. Analysis of the expression of cyclin D1 we know that the benign and malignant tumors have significant difference as compared to normal breast tissue(p<0.01), and the results of the expression of p27 show that it has significant difference in normal breast tissue and tumor tissue(p<0.01). According to the comparison between the situation of various types of tumors, infiltrating lobular carcinoma and ductal carcinoma have statistically significant difference about cyclin D1(p<0.05), p27 kip is statistically significant difference in mixed sample and intraductal carcinoma(p<0.05). There is a correlation between cyclin D1 and p27 kip. Through the above result we konw that although testing cyclin D1 and p27 kip through different methods to the test results are differences, but they guide significance to the diagnosis of canine breast tumor; cyclin D1 has reference value in early tumor diagnosis, the difference between intraductal carcinoma and invasive lobular carcinoma has reference meaning; Cyclin D1 and p27 kip jointly applied to the diagnosis of invasive ductal carcinoma has a value. |
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