Research On Expression Of PBK/TOPK In Lung Carcinoma And Normal Tissue

Background and ObjectivesLung cancer is increasingly detrimental to people's health. From the middle of the 20th century,the morbidity and mortality of lung cancer are growing rapidly worldwide. In most developed countries,lung cancer ranks first in males and second or third in females in commonly seen malignant tumors. Therefore, research on occurrence and development of mechanisms of lung cancer to prevent and control has important practical significance.PDZ-binding-kinase/T-LAK cell-originated protein kinase (PBK/TOPK), is a newly discovered silk-threonine kinase. Gaudet, etc. cloned a molecule in the Hela cell cDNA library with the PDZ domain of hDlg molecules combined, In the same year, another research team Abe, etc. found in a protein kinase similaried MAPKK molecules in the T-LAK cells, Later revealed that they were the same type molecule by sequence analysis, so which were unified named PDZ-binding-kinase/T-LAK-cell-originated protein kinase, referred to as PBK/TOPK. Initially found that it could bind with PDZ domain, it was reported in the literature that it could interact the tumor suppressor genes hDlg, P53 protein, raf protein and many other molecules and that could activate p38MAPK, Erk, JNK and other downstream molecules, as molecules involved in protein kinase signal transduction pathway. Past studies indicate that PBK /TOPK show low expression in the normal tissues besides bile duct epithelial cells, but high expression in several tumors, which led to researcher's attention. However, the research was rarely about lung cancer, studied the expression of PBK/TOPK by qualitative observation. Studies on the quantitative expression of PBK/TOPK in different kinds of lung carcinomas,its expression in lymph node metastases of lung carcinomas (including the qualitative and quantitative analyses) haven't been investigated.So, this study aims to detect the expression of PBK/TOPK protein and mRNA in lung cancer and normal human tissues.Quantitative analyses of the intensities of PBK/TOPK protein and mRNA were performed to revel the characteristics of PBK/ TOPK protein and mRNA level and research expression of PBK/TOPK normal human tissues.Methods1,20 cases of paraffin embeded normal lung tissues,20 cases of embryonic lung tissues,104 cases of paraffin embeded lung carcinomas and 46 cases of corresponding lymph node metastases were collected from the pathology department of Nanfang Hospital and the autopsy floor of Nanfang Hospital in 2008.10～2009.8. Lung carcinomas were analysed with respect to the relationship between PBK / TOPK expression and patient's sex,tumor general types,differentiation,lymph node metastasis and TNM stage.2,27 kinds of normal tissue,10 specimens of each type of organizations that include:lung, larynx, trachea, bronchus, esophagus, stomach, small intestine, large intestine, liver, kidney, adrenal gland, thyroid gland, brain, spinal cord, spleen, thymus, tonsils, heart, blood vessels, cartilage, muscle, testis, prostate, breast, ovarian, uterus,placenta were collected from the pathology department of Nanfang Hospital and the autopsy floor of Nanfang Hospital in 2008.10～2009.8. Investigation on expression of PBK/TOPK in different normal tissues.3,Tissue microarray technique was used to construct a tissue chip with 760 disks containing 20 cases of normal lung tissues,20 cases of embryonic lung tissues,104 cases of paraffin embeded lung carcinomas and 46 cases of corresponding lymph node metastases and to construct a tissue chip with 540 disks containing 27 kinds of normal tissue,10 specimens of each type of organizations.4,Immunohistochemical staining was used to detect PBK/TOPK protein expression in different kinds of lung carcinomas on the tissue chip with 760 disks and in different normal tissues on the tissue chip with 540 disks.Leica Q500MC image analysis system was used to dectect the intensity of PBK/TOPK protein expression. The relationship between PBK/TOPK protein expression in fifferent kinds of lung carcinomas was analysed.5,mRNA in situ hybridisation was used to detect PBK/TOPKmRNA expression in different kinds of lung carcinomas on the tissue chip with 760 disks and in different normal tissues on the tissue chip with 540 disks. Leica Q500MC image analysis system was used to dectect the intensity of PBK/TOPKmRNA expression. The relationship between PBK/TOPKmRNA expression in fifferent kinds of lung carcinomas was analysed.Results1,Immunohistochemical staining was used to dectect PBK/TOPK protein expression in lung carcinoma on the chip with 760 disks. (1) Embryonic alveolar epithelial cytoplasm of PBK/TOPK positive unit (PU) is less than the normal adult typeⅡalveolar epithelial cytoplasm of PBK/TOPK PU (P=0.000); (2) Lung squamous cell carcinoma, adenocarcinoma, large cell carcinoma cytoplasm of PBK/ TOPK PU values are greater than Embryonic alveolar epithelial and normal adult alveolar typeⅡepithelial cytoplasm of PBK/TOPK PU (P=0.000); Small cell lung cancer cytoplasm of PBK/TOPK PU is less than the normal adult typeⅡalveolar epithelial cytoplasm of PBK/TOPK PU (P=0.000); (3) Lung squamous cell carcinoma, lung adenocarcinoma,large cell lung cancer cytoplasm of PBK/ TOPK PU are greater than the small cell lung cancer cytoplasm of PBK/TOPK PU (P=0.000); (4) Lung squamous cell carcinoma, lung adenocarcinoma and large cell carcinoma lymph node metastasis of cancer cytoplasm of PBK/TOPK PU are greater than primary tumor cytoplasm of PBK/TOPK PU (t=23.938,P=0.000; t=17.688,P=0.000;t=6.582, P=0.000); Small cell lung cancer lymph node metastasis of cancer cytoplasm of PBK/TOPK PU and the primary tumor cytoplasm of PBK/ TOPK PU is basically the same (t=0.122,P=0.908);(5)PBK/TOPK PU in cancerous cells of lymph node metastases than in those withiout (t=3.064,P=0.003); (6) PBK/TOPK PU was bigger in TNM stageⅡ-Ⅳthan stageⅠ(t=3.002,P=0.003); (7) PBK/TOPK PU was not associated with patients'sex (t=1.226,P=0.223), general types (t=0.663,P=0.509) and differentiation (Squamous cell lung carcinoma: t=1.512,P=0.137; Lung adenocarcinoma:t=0.594,P=0.594).2,mRNA in situ hybridisation was used to dectect PBK/TOPKmRNA expression in lung carcinoma on the chip with 760 disks. (1) Embryonic alveolar epithelial cytoplasm of PBK/TOPKmRNA positive unit (PU) is less than the normal adult typeⅡalveolar epithelial cytoplasm of PBK/TOPKmRNA PU (P=0.000); (2) Lung squamous cell carcinoma, adenocarcinoma, large cell carcinoma cytoplasm of PBK/TOPKmRNA PU are greater than Embryonic alveolar epithelial and normal adult alveolar typeⅡepithelial cytoplasm of PBK/TOPKmRNA PU (P=0.000); Small cell lung cancer cytoplasm of PBK/TOPKmRNA PU is less than the normal adult typeⅡalveolar epithelial cytoplasm of PBK/TOPKmRNA PU (P=0.000); (3) Lung squamous cell carcinoma, lung adenocarcinoma,large cell lung cancer cytoplasm of PBK/TOPKmRNA PU are greater than the small cell lung cancer cytoplasm of PBK/TOPKmRNA PU (P=0.000); (4) Lung squamous cell carcinoma, lung adenocarcinoma and large cell carcinoma lymph node metastasis of cancer cytoplasm of PBK/TOPKmRNA PU are greater than primary tumor cytoplasm of PBK/TOPKmRNA PU (t=12.936,P=0.000;t=12.161, P=0.000;t=7.581, P=0.000); Small cell lung cancer lymph node metastasis of cancer cytoplasm of PBK/TOPKmRNA PU and the primary tumor cytoplasm of PBK/TOPKmRNA PU is basically the same (t=0.013,P=0.990);(5)PBK/TOPKmRNA PU in cancerous cells of lymph node metastases than in those withiout (t=3.665,P=0.000); (6) PBK/TOPKmRNA PU was bigger in TNM stageⅡ-Ⅳthan stageⅠ(t=3.050,P=0.003); (7) PBK/TOPKmRNA PU was not associated with patients'sex (t=0.594,P=0.554), general types (t=0.456,P=0.560) and differentiation (Squamous cell lung carcinoma:t=0.164,P=0.870; Lung adenocarcinoma:t=1.223,P=0.225).3,Immunohistochemical staining was used to dectect PBK/TOPK protein expression in normal tissue on the chip with 540 disks. There was not PBK/ TOPK protein expression in lung,larynx, trachea, bronchus, esophagus, stomach, small intestine, kidney, adrenal gland, thyroid, brain, spinal cord, spleen, thymus, tonsils, heart, blood vessels, cartilage, muscle, breast, testis, prostate, ovary, uterus and placenta. the glandular epithelial cells of Small intestinal expressed positive.4,mRNA in situ hybridisation was used to dectect PBK/TOPKmRNA expression in normal tissue on the chip with 540 disks. There was not PBK/ TOPK PBK/TOPKmRNA expression in lung,larynx, trachea, bronchus, esophagus, stomach, small intestine, kidney, adrenal gland, thyroid, brain, spinal cord, spleen, thymus, tonsils, heart, blood vessels, cartilage, muscle, breast, testis, prostate, ovary, uterus and placenta. the glandular epithelial cells of Small intestinal expressed positive. Conclusions1,The amount of PBK/TOPK expression differs in the cytoplasm of embryonic alveolar epithelial cells, normal adult alveolar typeⅡepithelial cells and lung cancer cells and increases orderly. The expression of PBK/TOPK differs in different types of lung carcinomas,which is comparatively higher in lung squamous cell carcinoma, adenocarcinoma and large cell carcinoma and lower in small cell carcinoma. The stronger the expression of PBK/TOPK,the easer lung carcinoma metastasizes.Cancer cells of lung squamous cell carcinoma, adenocarcinoma and large cell carcinoma with strong expression of PBK/TOPK in the cytoplasm are one of the important hallmarks of cells with distinct potential of metastasis.2,The amount of PBK/TOPKmRNA expression differs in the cytoplasm of embryonic alveolar epithelial cells, normal adult alveolar typeⅡepithelial cells and lung cancer cells and increases orderly. The expression of PBK/TOPKmRNA differs in different types of lung carcinomas,which is comparatively higher in lung squamous cell carcinoma, adenocarcinoma and large cell carcinoma and lower in small cell carcinoma. The stronger the expression of PBK/TOPKmRNA,the easer lung carcinoma metastasizes.Cancer cells of lung squamous cell carcinoma, adenocarcinoma and large cell carcinoma with strong expression of PBK/ TOPKmRNA in the cytoplasm are one of the important hallmarks of cells with distinct potential of metastasis.3,PBK/TOPK protein expressed in few normal tissues and didn't express in most normal tissues. PBK/TOPK showed no expression in adult lung tissues and in embryonic tissues, but high expression in lung cancer tissues, which indicates PBK/ TOPK is closely related to lung cancer formation.4,PBK/TOPKmRNA expressed in few normal tissues and didn't express in most normal tissues. PBK/TOPKmRNA showed no expression in adult lung tissues and in embryonic tissues, but high expression in lung cancer tissues, which indicates PBK/TOPK is closely related to lung cancer formation.