| IntroductionInhibitor of apoptosis protein (IAP) family is important to apoptosis. Its central mechanism is direct or indirect inhibition of Caspase. Survivin is a novel number of IAP families. Survivin is the most powerful inhibitor of apoptosis as it inhibits Caspase - 3 and Caspase - 7 , the terminal effecter of apoptotic pathway.Survivin protein is composed of 142 amino acids; its molecular weight is about 16.4kD. Survivin preserves the most important function region, BIR2 ( baculovirus IAP repeat) , of IAP molecule. Survivin gene is located on 17q25, 75 -130 kb in length. It includes 4 extrons and 3 introns. Survivin expression is seen in the tissue of embryo and fetus, but not in terminal differentiated adult tissues except for endometrium, thymus and placenta. Survivin is extensively expressed in almost all the malignant tissues, such as lung cancer, colo-rectal cancer, hepatic cell carcinoma, breast cancer, etc. And the expression of Survivin is cell - cycle dependent. Survivin dont express in G1 phase, and over express in G2/M phase. There is a relationship between Survivin expression and cell multiplication. Many evidence suggested that Survivin was important for tumorigenesis. It is possible to use genetic therapy targeting to Survivin for anti - tumor therapy through promoting apoptosis of tumor cells due to specific distributionof Survivin.Pancreatic carcinoma is a common malignant tumor of the digestive system. It's difficult to make an early diagnosis and the excision rate is limited. The exact tumorigenesis of pancreatic carcinoma is still unknown at present. There are only a few reports are about Survivin and pancreatic carcinoma. The relationship between Survivin and Bel -2 and p53 is unclear. We detect Survivin expression and its relationship to Bel -2 and p53 genes. Therefore, Survivin may provide a new adjuvant therapeutic method for pancreatic cancer.Materials and MethodsPatients and specimen-.Pancreatic carcinoma specimens were selected from paraffin wax embedded specimens with full clinicopathological data. All the patients underwent operations in the 1st and 2nd affiliated hospital of China Medical University and the postoperative pathologic diagnosis was pancreatic carcinoma. There were 26 men and 24 women, with mean age of (57. 7 +7.4) years old. Histological differentiation; high 18 cases, moderate 20 cases, low 12 cases. There were 26 cases with local lymph nodes metastasis and 24 cases without. The clinical stage was divided according to the standard of UICC (1997) : stage I 14 cases, stage II 12 cases, stage HI 14 cases, stage IV 10 cases. And 5 cases of chronic pancreatitis tissues and 12 cases of normal pancreatic tissues were studied as control.Main reagents;Goat anti - human Survivin monoclonal antibody ( Santa Cruz Corp. , USA) , mouse anti - human Bel -2 and p53 monoclonal anti-bodies (Maixin Corp. ) .Immunohistochemical staining:Specimens from the paraffin wax blocks were cut into 5|xm sections. The first section was routinely stained with hematoxylin - eosin for histological diagnosis and differentiation. Immunohistochemical staining for Survivin, Bel - 2, and p53 was performed with a standard avidin - biotin - peroxidase complex detection kit. The sections were dewaxed and rehydrated; antigen repair 1.5 min through high temperature and pressure method with 0. 01M PH6. 0 citrate buffer solution. Endogenous peroxidase activity and nonspecific bindings were blocked by incubation with 3% hydrogen peroxide and non - immune serum, respectively. Slides were then incubated sequentially with primary antibodies ( dilution 1: 200 for Survivin, Bel - 2 and p53 are instant antibodies) overnight at 4c with a biotinylated secondary antibodies ( rabbit anti - goat for Survivin, rabbit and - mouse for Bcl - 2 and p53) for 30 min and with SP complex for 30 min. Then the chromogen DAB test was performed to be able to localize the positive staining by microscope sequentially. The sections were counterstained with hematoxylin, dehydrated in graded eth...
|
PDF Full Text Request