Study On The Expression Of Aldolase A And Cytokeration19in Sheep Pulmonary Adenomatosis

Sheep pulmonary adenomatosis(SPA) is caused by jaagsiekte sheep retrovirus(JSRV).The virus induces neoplastic transformation of alveolar type Ⅱ pneumocytes and clara cells in sheep lung.Sheep pulmonary adenomatosis has a long incubation period.Frothy mucoid fluid accumulates within the respiratory tract and may be seen leaking out from the nostrils after raising the back and lowering the head of the sheep.Diagnosis of sheep pulmonary adenomatosis is usually possible only in the late stage of the disease.The neoplastic growth in the lungs may be not recognized until postmortem examination.Sheep pulmonary adenomatosis has been found to be clinically and histologically similar to bronchiolo-alveolar carcino-ma.The possibility of experimental induction of the tumor in animals makes it a good model for the study of oncogenesis as well as possibilities for diagnostics and therapy of lung tumors. Studying the differential expression of proteins in the neoplastic tissues of sheep pulmonary adenomatosis may lead to the discovery of protein markers.A.Kycko found that aldolase A and cytokeratin19were overexpressed in sheep pulmonary adenomatosis.In the foreign study the protein profiles of neoplastic and non-neoplastic sheep lung tissues were examined for the identification of proteins overexpressed in sheep pulmonary adenomatosis.Two dimensional electrophoresis was performed using gel strips with immobilized pH gradient3-10.In two spots demonstrating over3-fold higher expression in sheep pulmonary adenomatosis proteomes,single proteins:cytokeratin19and aldolase A.The expression of aldolase A and ckl9in sheep pulmonary adenomatosis and non-neoplastic lung tissue were detected by immunohistochemistry and real-time PCR in this study.The positive unit and mean optical density in aldolase A and ck19were used to analyze quantitatively by Image-Pro Plus image analysis system. Results revealed that positive immunostaining of aldolase A and ck19visible in cytoplasm of the bronchial and individual alveolar epithelial cells in non-neoplastic lung tissue,as well as the cytoplasm of neoplastic cells in sheep pulmonary adenomatosis.The mean optical density of aldolase A in sheep pulmonary adenomatosis was significantly higher than in non-neoplastic lung tissue(P<0.05).The mean optical density of aldolase A in sheep pulmonary adenomatosis was about3.7times as much as the mean optical density of aldolase A in non-neoplastic lung tissue. The mean optical density of aldolase A in sheep pulmonary adenomatosis was 0.352500±0.0335339, The mean optical density of aldolase A in non-neoplastic lung tissue was0.094480±0.0067188.The positive unite(PU)of aldolase A in sheep pulmonary adenomatosis was very significantly higher than in non-neoplastic lung tissue(P<0.01).The positive unite of aldolase A in sheep pulmonary adenomatosis was about2.6times as much as the positive unite of aldolase A in non-neoplastic lung tissue. The positive unite of aldolase A in sheep pulmonary adenomatosis was20.7225±0.08261,The positive unite of aldolase A in non-neoplastic lung tissue was7.8125±0.03594. The mean optical density of ck19in sheep pulmonary adenomatosis was significantly higher than in non-neoplastic lung tissue(P<0.05).The mean optical density of ck9in sheep pulmonary adenomatosis was about3.8times as much as the mean optical density of ck19in non-neoplastic lung tissue. The mean optical density of ck19in sheep pulmonary adenomatosis was0.365150±0.0322141, The mean optical density of ck19in non-neoplastic lung tissue was0.096075±0.0069916.The positive unite(PU)of ck19in sheep pulmonary adenomatosis was very significantly higher than in non-neoplastic lung tissue(P<0.01).The positive unite of ck19in sheep pulmonary adenomatosis was about2.7times as much as the positive unite of ck19in non-neoplastic lung tissue.The positive unite of ck19in sheep pulmonary adenomatosis was20.4625±0.14523,The positive unite of ck19in non-neoplastic lung tissue was7.5275±0.33689. The expression of aldolase A in sheep pulmonary adenomatosis and control group were detected by real-time PCR, and the result was analyzed by2-△△CTquantitative method.The aldolase A of sheep pulmonary adenomatosis was about2.22times as much as the aldolase A of control group.The results shows that the overexpression of aldolase A and ck19in sheep pulmonary adenomatosis.