Objective To investigate the invasion ability of Panc-1 cells in vivo and in vitro af ter being t ransfected with tissue factor pathway inhibitor 2 gene ( TFPI-2) . Methods The expression vector pEGFP-C1-TFPI-2 was transfected into human pancreatic cancer line Panc-1 cells by using liposome. TFPI-2 mRNA and protein of transfected and nontransfected cells were detected by reverse t ranscription-polymerase chain reaction (RT-PCR) and Western blot respectively. The tumor cells invasive behavior of t ransfected ( Panc-1-TFPI-2) and nontransfected ( Panc-1-V and Panc-1-P) cells were assessed in vitro through Boyden Chamber method. The transfected and nontransfected cells were implanted into nude mice to observe it s growth and metastasis in vivo. Results Expressions of mRNA and protein of TFPI-2 were confirmed in transfected cells. Af ter TFPI-2 t ransfection , the number of Panc-1-TFPI-2 , Panc-1-V and Panc-1-P cells passing through membrane of Boyden Chamber were 24. 4 ±3. 5 ,61. 3 ±4. 1 and 60. 2 ±3. 9 , respectively. The number of TFPI-2-expressing cells to t raverse a Matrigel-coated membrane was obviously decreased compared with that of non-expressing cells , the invasion ability was lower than that before transfection in vitro. The subcutaneous tumor volume of the Panc-1-TFPI-2 group was (438. 0 ±69. 8) mm3 , the Panc-1-V group was (852. 0 ±102. 9) mm3 and the Panc-1-P group was (831. 0 ±78. 1) mm3 , P lt; 0. 05. The metastasis to liver and lung and muscular invasion occurred in the Panc-1-V group and the Panc-1-P group. There were no muscular invasion and metastatic lesions in the Panc-1-TFPI-2 group. Conclusion TFPI-2 gene expression may obviously inhibit the invasion ability of pancreatic cancer cells in vitro and in vivo , which provides an experimental basis for the treatment of human pancreatic cancer by gene therapy.
ObjectiveTo explore the values of CA19-9, CA242, CEA, and CA125 single or combined detection on clinical diagnosis and prognosis for patients with pancreatic cancer. MethodsSerum tumor markers CA199, CA242, CEA, and CA125 of 63 patients with pancreatic cancer, 33 patients with cancer of bile duct, and 27 patients with benign pancreatic disease were detected, and those patients were followed up after operation. ResultsThe levels of CA19-9, CA242, CEA, and CA125 in patients with pancreatic cancer were significantly higher than those in patients with benign pancreatic disease and cancer of bile duct (Plt;0.05). The sensitivity of CA19-9 alone was the highest in the four tumor markers for the patients with pancreatic cancer 〔79.4% (50/63)〕, but the specificity (61.9%) was lower than that of CA242 (83.3%) and CEA (80.0%). The specificity of combined detection of CA199+CA242+CEA was the highest 〔93.3% (56/60)〕. The level of CA19-9 in carcinoma of body/tail of pancreas was significantly higher than that of carcinoma of pancreas head or whole pancreas (Plt;0.05). The serum levels of CA19-9 and CA242 in patients with stage Ⅳ were significantly higher than those in stage Ⅰ or Ⅱ/Ⅲ (Plt;0.05). Fifteen patients were lost to follow up, 48 patients were followed up 2－12 months with an average 6 months. The levels of CA242 and CA199 in patients with pancreatic cancer on 0.5 month and 3 months after operation were lower than those before operation (Plt;0.05). ConclusionsSingle detection of CA19-9 can improve the diagnostic sensitivity, and combined detection of tumor markers CA199+CA242+CEA can improve the diagnostic specificity. CA19-9 or CA242 is a valuable marker for evaluating treatment effects and estimating prognosis.
Objective To introduce the research progress in the effect of chemotherapeutic resistance of metabolic enzymes of gemcitabine to pancreatic cancer.Methods Recent literatures about metabolic enzymes that played key roles in mediating gemcitabine chemotherapeutic resistance of pancreatic cancer were collected and reviewed. Results The metabolic enzymes of gemcitabine, such as hENT1, dCK, RRM1 and CDA, were closely related to chemotherapeutic resistance of pancreatic cancer. The relationship between the single nucleotide polymorphism of metabolic enzymes and the resistance to gemcitabine remained to be clarified. Conclusion Multiple factors are involved in the mechanism of chemotherapeutic resistance of pancreatic cancer to gemcitabine, which needs further research.
ObjectiveTo summary the detection methods of circulating tumor cells (CTCs) in pancreatic cancer patients and its clinical application. MethodsRelated domestic and foreign literatures were reviewed. ResultsPancreatic cancer is one of the common malignant tumors in the world. The early diagnosis rate is low, the incidence of local invasion and metastasis is high, and the prognosis is very poor. The CTCs is one of the important causes of postoperative recurrence and metastasis, its detection methods based on immunocytochemistry (ICC) and reverse transcription polymerase chain reaction (RT-PCR). ConclusionsDetection of CTCs, regarding as a "real-time liquid biopsy", it has a high application value in the early diagnosis, treatment, prognosis and effect evaluation of pancreatic cancer, and it has become research frontier and focus.
Objective To investigate the expressions of CXCR4 and β-catenin in pancreatic cancer, explore the relationship between them, and explore the possible biomarkers about invasion and metastasis of pancreatic cancer. Methods Forty-eight samples of pancreatic cancer and 20 samples of normal pancreas tissues were selected. The expressions of CXCR4 and β-catenin were examined by the immunohistological technique. Spearman, Chi-square， and rank test were used to analyze the relation between the protein expressions and clinical characteristics. Survival analysis was evaluated by Kaplan-Meier product limit method and Log-rank test. Variables were evaluated by Cox proportional hazards analysis. The size of test was 0.05. Results The positive expression rates of CXCR4 and β-catenin in pancreatic cancer tissues were 85.4% (41/48) and 75.0% (36/48), respectively. Co-expression rate of CXCR4 and β-catenin was 70.8% (34/48). There were significant differences between various CXCR4 staining and lymph node metastasis and TNM stage (P=0.012, 0.005, respectively). β-catenin positive expression was associated with lymph node metastasis (P=0.047). However, abnormal β-catenin positive expression could not determine the clinical survival. Kaplan-Meier estimated curves suggested that clinical prognosis was poor for patients with CXCR4 expression. Multivariate analysis showed that CXCR4, late TNM stage, and lymph node metastasis were independent prognostic factors for pancreatic cancer. Conclusions Both CXCR4 and β-catenin abnormally express in pancreatic cancer. CXCR4 may be an important marker for pancreatic cancer progression.
Objective To observe the expression of Galectin-3 and Galectin-1 in pancreatic cancer and explore the relationship between the expression and pathological grading. Methods Forty specimens of pancreatic carcinoma tissue and thirty-one specimens of normal pancreas tissue were selected, which were confirmed by surgical resection and pathology from 2002 to 2009. The expression of Galectin-3 mRNA and Galectin-1 mRNA in pancreatic cancer cell lines SW1990, PANC-1 and ASPC -1 was detected by means of reverse transcriptase-polymerase chain reaction; the expression of Galectin-3 protein and Galectin-1 protein in SW1990, PANC-1 and ASPC-1 was detected by means of immunocytochemistry; the expression of Galectin-3 protein and Galectin-1 protein in pancreatic cancer and normal pancreatic tissue was detected by means of immunohistochemistry. Results In SW1990, PANC-1 and ASPC-1, Galectin-3 mRNA signal and protein were detected, but no Galectin-1 mRNA signal or protein was detected. There was no expression of Galectin-3 protein or Galectin-1 protein in the 31 specimens of normal pancrease tissue, while there were Galectin-3 protein and Galectin-1 protein expressed in the 40 specimens of pancreatic cancer tissue. In the 40 specimens of pancreatic cancer tissue, the expression of Galectin-3 protein was observed in pancreatic cancer cells, but not in fibroblasts or matrix cells around the cancer mass; while the expression of Galectin-1 protein was observed in fibroblasts and matrix cells around the cancer mass, but not observed in pancreatic cancer cells. There was no significant association between the expression of Galectin-3 protein in pancreatic cancer and pathological grading (P>0.05); while the expression of Galectin-1 protein in pancreatic cancer was related to the pathological grading, and the expression of Galectin-1 protein was significant higher in poorly differentiated tumors than that in well/moderately differentiated tumors (P<0.05). Conclusions Galectin-3 or Galectin-1 is not expressed in normal pancreases; Galectin-3 is expressed in pancreatic cancer cells; Galectin-1 is expressed in fibroblasts and matrix cells around the cancer mass. The expression of Galectin-1 is related with the differentiation of pancreatic cancer.
ObjectiveTo study the expression of lipid associated with neutrophil gelatinase associated lipocalin (NGAL) in nude mice orthotopic pancreatic cancer tissues and the relationship between the occurred and development of pancreatic cancer. MethodsThe expressions of NGAL mRNA and protein of pancreatic cancer tissues and their adjacent tissues, and normal pancreatic tissues in nude mice were detected by using RT-PCR and immunohistochemical methods. ResultsThe expressions of NGAL mRNA in pancreatic cancer tissues and adjacent tissues were significantly higher than that in normal pancreatic tissues (P < 0.05), and the expression of NGAL mRNA in pancreatic carcinoma tissues was significantly higher than that in para carcinoma tissues (P < 0.05). The strong positive expression rate of NGAL protein in pancreatic carcinoma tissues was significantly higher than thoes in para carcinoma tissues and normal pancreatic tissues (P < 0.05). ConclusionsNGAL is highly expressed in pancreatic cancer tissues, and NGAL may be an important regulatory factor in the development of pancreatic cancer.
Objective To observe the effect of epidermal growth factor (EGF) on the proliferation, adhesion, invasiveness and the activation of nuclear factor-κB (NF-κB), matrix metalloproteinases (MMPs) expression and explore related mechanisms in pancreatic cancer cells. Methods Cell invasion assay, proliferation assay and adhesion assay were used to examine the proliferation, adhesion and invasiveness of pancreatic cancer cells, respectively. NF-κB activity was detected by electrophoretic mobility shift assay (EMSA), and MMPs protein and mRNA expressions were investigated by gelatin zymography, Western blot and reverse transcriptase-polymerase chain reaction (RT-PCR). Results EGF increased the invasiveness of pancreatic cancer cell in a dose-dependent manner (P＜0.05), but did not affect cell proliferation or adhesion. The expressions of MMP-9 mRNA and protein significantly increased after induction by EGF and were highest when EGF concentration was 50 ng/ml, while there was no effect on the expressions of MMP-2 mRNA and protein. Furthermore, NF-κB activity increased with increased concentration of EGF in a concentration-dependent manner (P＜0.05). In addition, NF-κB activity and the expressions of MMP-9 mRNA and protein by pretreatment with both pyrrolidine dithiocarbamate (PDTC) and EGF decreased when compared that by pretreatment with EGF alone. The invasiveness of pancreatic cancer cell by pretreatment with both PDTC and EGF decreased when compared that by pretreatment with EGF alone and nothing (P＜0.05).Conclusion The findings indicate that the NF-κB-mediated MMP-9 induction is essential for EGF-induced invasiveness in pancreatic cancer cells, which can be inhibited by PDTC.
Objective To summary the principle of magnetic resonance spectroscopy imaging and its application progress in diagnosis and differential diagnosis of pancreatic cancer. Methods The newest related literatures of home and abroad were collected and reviewed. Results Magnetic resonance spectroscopy imaging was a technology using the magnetic resonance phenomena and chemical shift phenomena to measure molecular organization. The spectroscopy most commonly used in clinical and scientific research includes 1H， 31P， and 23Na. Conclusion Magnetic resonance spectroscopy as the only approach to noninvasive quantitative provding biochemical information in vivo， has an important significance to the diagnosis and differential diagnosis of pancreatic cancer.
【Abstract】Objective To analyze the function of BAG3 in antiapoptosis and chemotherapy resistance induction process of pancreatic cancer.Methods The expressions of BAG-3 in pancreatic cancerous tissues of patients with chemotherapy and those without chemotherapy before resection were determined by immunohistochemistry. The expression difference of BAG-3 protein 18 hours after cultured with chemotherapy drugs (concentration of drugs: 5-FU 50 μg/ml, MMC 0.5 μg/ml, EADM 1.5 μg/ml) of 3 pancreatic cancer cell lines （MIACaPa-2, PANC-1, SW1990） was measured through Western blotting method.Results The median positive rate of pancreatic cancer tissue from patients accepted chemotherapy before resection was higher than those not accepted chemotherapy, but there wasn’t significant difference. Eighteen hours after cultured with drugs, the level of BAG-3 of this three cell lines had significant increased compared with control group (P＜0.05). Conclusion Chemotherapy induces elevation of BAG-3 expression of pancreatic cancer. The upregulate of BAG-3 may associate with the chemotherapy resistance induced by drugs.