| Extremely high levels of pancreatic digestive malignant tumor incidence in the world and our country radiate a rising trend year by year. A high degree of malignancy of pancreatic cancer, symptoms hidden, when the typical clinical manifestations difficult to treat, the current lack of clinical lack of a means of early diagnosis. Further study the pathogenesis of pancreatic cancer, to find the disease early diagnosis, treatment of pancreatic carcinoma has become a key issue. In the pathogenesis of cancer research, and gradually and orderly genetic changes caused by oncogene activation and inactivation of tumor suppressor genes in human cancer is considered an important mechanism of the molecular biology. However, as the cancer itself is a complex study of the biological behavior of in-depth, simple genetic changes can not fully explain the whole process of tumor pathogenesis, the genetic modification of epigenetic gene expression may have an important impact. DNA methylation is an important research content of epigenetics, DNA methyltransferase 1 (DNA methyltransferase 1) was the first one being the transfer of cloned DNA methylation enzymes, currently considered the role of DNMT1 to maintain CpG island methylation technology, as well as human cancer tumor suppressor gene silencing-related abnormalities.In this study, using immunohistochemistry and western blot method, combined with a complete clinical data on pancreatic cancer and normal tissue in the pancreas, the expression of DNMT1 to conduct research and analysis of DNMT1 and clinical pathological factors in order to explore it in Pancreatic Cancer Expression and its significanceMaterials and Methods一. Materials 47 cases of pancreatic cancer and 22 cases of normal tissues were from the First Affiliated Hospital of China Medical University from 2004 to 2009 archived wax blocks of surgical resection. Organizations have been 10% formalin-fixed, conventional paraffin-embedded, cut into 4μm thick consecutive sections prepared by immunohistochemical and pathological analysis.9 cases of pancreatic cancer and normal tissue from the First Clinical Hospital of China Medical University Gastrointestinal Pancreatic Surgery in 2008～2010 surgical resection specimens. Surgery immediately after excision from the primary tumor and surrounding normal pancreatic tissue were obtained from a diameter of about 0.5cm tissue placed on the EP tube,-70℃refrigerator spare.二. Methods(一)Immunohistochemistry:DNMT1 expression detected using streptavidin-biotin-peroxidase immunohistochemical method (sabc law). Results found:DNMT1 protein was expressed in the nucleus, light microscope, each slice select five high power field, high power field for each count of 100 tumor cells, small nuclei stained yellow or yellow as positive, stained cells were observed under a microscope ratio, large than 50% positive, less than 50% of the negative.(二)Western blot Detection of DNMT1 in normal pancreatic tissue and pancreatic cancer. Get fresh samples (100 mg) plus 5-fold lysis buffer, Shredded, homogenized ultrasonic treatment, high-speed cryogenic centrifugal supernatant protein extraction, electrophoresis, transfer, after the closure plus the corresponding primary antibody, anti-, and two color reagents detection. The experimental results using analysis software Quantity One gel electrophoresis determination of gray values as quantitative indicators to determine the relative content of DNMT1 protein.三.Statistical analysisUsing SPSS for windows 13.0 software, DNMT1 expression and clinical pathological features of the relationship between the use of tests. Western blot results of the gray values divided into pancreatic cancer group and normal group paired t test. P<0.05 was significant. 一.Expression of DNMT1 in pancreatic carcinomas and normal tissuesWestern Blot results:DNMT1 in pancreatic cancer and normal tissues have different levels of expression in pancreatic cancer tissues was significantly higher than the corresponding normal tissues of the pancreas二.the relationship of DNMT1 and clinic pathology factorsPancreatic cancer, DNMT1-positive rate was 70.2%(33/47), its expression with tumor clinical staging, differentiation degree and lymph node metastasis (P<0.05) correlated with age, sex has nothing to do (P> 0.05)Conclusion1.DNMT1 expression in pancreatic cancer tissues was significantly higher than in normal pancreatic tissue.2. Pancreatic, DNMT1 expression and tumor clinical staging, differentiation degree and lymph node metastasis correlated with age, sex has nothing to do... |
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