AIM:To investigate the inhibitory effects of RNA interference (RNAi) on expression of matrix metalloproteinase-2 (MMP-2) gene and invasiveness and adhesion of human pancreatic cancer cell line,BxPC-3.METHODS:RNAi was ...AIM:To investigate the inhibitory effects of RNA interference (RNAi) on expression of matrix metalloproteinase-2 (MMP-2) gene and invasiveness and adhesion of human pancreatic cancer cell line,BxPC-3.METHODS:RNAi was performed using the vector (pGPU6)-based small interference RNA (siRNA) plasmid gene silence system to specifically knock down MMP-2 expression in pancreatic cancer cell line,BxPC-3. Four groups of different specific target sequence in coding region of MMP-2 and one non-specific sequence were chosen to construct four experimental siRNA plasmids of pGPU6-1,pGPU6-2,pGPU6-3 and pGPU6-4,and one negative control siRNA plasmid of pGPU6 (-). MMP-2 expression was measured by reverse transcription polymerase chain reaction (RT-PCR) and Western blot. Cell proliferation and apoptosis were examined by methyl thiazolyl tetrazolium (MTT) and flow cytometry,respectively. The abilities of adhesion and invasion were detected by cell adhesion assay and cell invasion assay using Transwell chambers.RESULTS:The expression of MMP-2 was inhibited and the inhibitory effects of different sequence varied. pGPU6-1 group had the most efficient inhibitory effect,followed by pGPU6-2 and pGPU6-3 groups.Invasiveness and adhesion were more significantly reduced in pGPU6-1,pGPU6-2 and pGPU6-3 groups as compared with pGPU6 (-) and blank control groups. However,no difference concerning cell proliferation and apoptosis was observed after transfection between experiment groups and control groups.CONCLUSION:RNAi against MMP-2 successfully inhibited the mRNA and protein expression of MMP-2 in the pancreatic cancer cell line,BxPC-3,leading to a potent suppression of tumor cell adhesion and invasion without affecting cell proliferation and apoptosis. These findings suggest that the RNAi approach towards MMP-2 may be an effective therapeutic strategy for the clinical management of pancreatic tumor.展开更多
A case of intraductal papillary neoplasm of the bile duct (IPNB) arising in a patient with hepatitis B-related liver cirrhosis with hepatocellular carcinoma (HCC) is reported. A 76-year-old man was admitted to our hos...A case of intraductal papillary neoplasm of the bile duct (IPNB) arising in a patient with hepatitis B-related liver cirrhosis with hepatocellular carcinoma (HCC) is reported. A 76-year-old man was admitted to our hospital with recurrent HCC. Laboratory data showed that levels of carcinoembryonic antigen and carbohydrate antigen 19-9 were elevated. He died of progressive hepatic failure. At autopsy,in addition to HCCs,an intraductal papillary proliferation of malignant cholangiocytes with fibrovascular cores was found in the dilated large bile ducts in the left lobe,and this papillary carcinoma was associated with an invasive mucinous carcinoma (invasive IPNB). Interestingly,extensive intraductal spread of the cholangiocarcinoma was found from the reactive bile ductular level to the interlobular bile ducts and septal bile ducts and to the large bile ducts in the left lobe. Neural cell adhesion molecule,a hepatic progenitor cell marker,was detected in IPNB cells. It seems possible in this case that hepatic progenitor cells located in reactive bile ductules in liver cirrhosis may have been responsible for the development of the cholangiocarcinoma and HCC,and that the former could have spread in the intrahepatic bile ducts and eventually formed grossly visible IPNB.展开更多
AIM： To investigate the effects of integrin-linked kinase （ILK） on gastric cancer cells both in vitro and in vivo. METHODS： ILK small interfering RNA （siRNA） was transfected into human gastric cancer BGC-823 cel...AIM： To investigate the effects of integrin-linked kinase （ILK） on gastric cancer cells both in vitro and in vivo. METHODS： ILK small interfering RNA （siRNA） was transfected into human gastric cancer BGC-823 cells and ILK expression was monitored by real-time quan- titative polymerase chain reaction, Western blotting analysis and immunocytochemistry. Cell attachment, proliferation, invasion, microfilament dynamics and the secretion of vascular endothelial growth factor （VEGF） were also measured. Gastric cancer cells treated with ILK siRNA were subcutaneously transplanted into nude mice and tumor growth was assessed. RESULTS： Both ILK mRNA and protein levels were significantly down-regulated by ILK siRNA in human gastric cancer cells. This significantly inhibited cell attachment, proliferation and invasion. The knockdown of ILK also disturbed F-actin assembly and reduced VEGF secretion in conditioned medium by 40% （P 〈 0.05）. Four weeks after injection of ILK siRNA-transfected gastric cancer cells into nude mice, tumor volume and weight were significantly reduced compared with that of tumors induced by cells treated with non-silencing siRNA or by untreated cells （P 〈 0.05）. CONCLUSION： Targeting ILK with siRNA suppresses the growth of gastric cancer cells both in v/tro and /n vivo. ILK plays an important role in gastric cancer progression.展开更多
Objective: To observe the changes of plasma concentrations of endotoxin, soluble intercellular adhesion molecule 1, tumor necrosis factor α, and urinary microalbumin in children undergoing cardiac procedure and to st...Objective: To observe the changes of plasma concentrations of endotoxin, soluble intercellular adhesion molecule 1, tumor necrosis factor α, and urinary microalbumin in children undergoing cardiac procedure and to study the effects of cardiopulmonary bypass (CPB) on the injury or activation of endothelial cells and vascular permeability. Methods: Twenty children undergoing cardiac operation with CPB were selected in the study. Plasma concentrations of endotoxin, soluble intercellular adhesion molecule 1, tumor necrosis factor α, and urinary microalbumin were measured after anesthetic induction (baseline), bypass for 20 minutes, at the end of CPB, and at 2, 4, and 18 h after the end of CPB. Results: The plasma concentrations of endotoxin, soluble intercellular adhesion molecule 1, and urinary microalbumin began to increase at 2 h after the end of CPB, and remained higher than that of the baseline, while the concentration of tumor necrosis factor α increased only at the end CPB and at 2 h after the end of CPB. Conclusion: Cardiopulmonary bypass can induce inflammatory response, resulting in the activation or injury of vascular endothelial cells, and can increase the vascular permeability.展开更多
OBJECTIVE: To investigate the reasons for the rarity of metastases in skeletal muscle. METHODS: By injecting tumor cells (Walker256 rat carcinosarcoma) through the iliac artery (experimental group) and the tail vein (...OBJECTIVE: To investigate the reasons for the rarity of metastases in skeletal muscle. METHODS: By injecting tumor cells (Walker256 rat carcinosarcoma) through the iliac artery (experimental group) and the tail vein (control group), animal models of blood-borne metastases were established. The quadriceps femoris muscle and lungs were observed grossly and microscopically. Immunohistochemistry was applied to investigate the expression of vascular cell adhesion molecule-1 (VCAM-1) in the microvascular endothelium of these organs. Primary culture of rat skeletal muscle cells was established and conditioned medium (MCM) was collected. Effects of MCM on several tumor cell lines and the biochemical characteristics of skeletal muscle delivered tumor factor(s) were tested by MTT assay. Apoptosis and morphological examination were carried out to investigate the antitumor mechanisms of MCM. RESULTS: In the experimental group, there were no definite metastases observed in muscle cells. In the control group, lung metastases were present in the lungs of all rats that were sacrificed at the 14th day or died spontaneously (17 rats in all). There was no significant difference between the increase in VCAM-1 in quadriceps femoris muscle 7 days after iliac artery injection and that in lungs 7 days after tail vein injection (P > 0.05). In vitro studies showed that the proliferation of tumor cell lines of mouse SP2/0 myeloma, rat Walker256 carcinosarcoma or human chronic granulocytic leukemia K562, human acute lymphatic leukemia HL-60, LS-174-T colon adenocarcinoma, PC3-M prostatic carcinoma and lung giant cell carcinoma with different metastatic potency (PLA801-C with low metastatic potency, PLA801-D with high metastatic potency) was significantly inhibited when cultured with MCM (P展开更多
基金Supported by Tiantan Hospital Scientific Project Grant Fund
文摘AIM:To investigate the inhibitory effects of RNA interference (RNAi) on expression of matrix metalloproteinase-2 (MMP-2) gene and invasiveness and adhesion of human pancreatic cancer cell line,BxPC-3.METHODS:RNAi was performed using the vector (pGPU6)-based small interference RNA (siRNA) plasmid gene silence system to specifically knock down MMP-2 expression in pancreatic cancer cell line,BxPC-3. Four groups of different specific target sequence in coding region of MMP-2 and one non-specific sequence were chosen to construct four experimental siRNA plasmids of pGPU6-1,pGPU6-2,pGPU6-3 and pGPU6-4,and one negative control siRNA plasmid of pGPU6 (-). MMP-2 expression was measured by reverse transcription polymerase chain reaction (RT-PCR) and Western blot. Cell proliferation and apoptosis were examined by methyl thiazolyl tetrazolium (MTT) and flow cytometry,respectively. The abilities of adhesion and invasion were detected by cell adhesion assay and cell invasion assay using Transwell chambers.RESULTS:The expression of MMP-2 was inhibited and the inhibitory effects of different sequence varied. pGPU6-1 group had the most efficient inhibitory effect,followed by pGPU6-2 and pGPU6-3 groups.Invasiveness and adhesion were more significantly reduced in pGPU6-1,pGPU6-2 and pGPU6-3 groups as compared with pGPU6 (-) and blank control groups. However,no difference concerning cell proliferation and apoptosis was observed after transfection between experiment groups and control groups.CONCLUSION:RNAi against MMP-2 successfully inhibited the mRNA and protein expression of MMP-2 in the pancreatic cancer cell line,BxPC-3,leading to a potent suppression of tumor cell adhesion and invasion without affecting cell proliferation and apoptosis. These findings suggest that the RNAi approach towards MMP-2 may be an effective therapeutic strategy for the clinical management of pancreatic tumor.
文摘A case of intraductal papillary neoplasm of the bile duct (IPNB) arising in a patient with hepatitis B-related liver cirrhosis with hepatocellular carcinoma (HCC) is reported. A 76-year-old man was admitted to our hospital with recurrent HCC. Laboratory data showed that levels of carcinoembryonic antigen and carbohydrate antigen 19-9 were elevated. He died of progressive hepatic failure. At autopsy,in addition to HCCs,an intraductal papillary proliferation of malignant cholangiocytes with fibrovascular cores was found in the dilated large bile ducts in the left lobe,and this papillary carcinoma was associated with an invasive mucinous carcinoma (invasive IPNB). Interestingly,extensive intraductal spread of the cholangiocarcinoma was found from the reactive bile ductular level to the interlobular bile ducts and septal bile ducts and to the large bile ducts in the left lobe. Neural cell adhesion molecule,a hepatic progenitor cell marker,was detected in IPNB cells. It seems possible in this case that hepatic progenitor cells located in reactive bile ductules in liver cirrhosis may have been responsible for the development of the cholangiocarcinoma and HCC,and that the former could have spread in the intrahepatic bile ducts and eventually formed grossly visible IPNB.
基金Supported by The grants from the Department of Anesthesiology and Intensive Care of Changhai Hospital,Shanghai,China
文摘AIM： To investigate the effects of integrin-linked kinase （ILK） on gastric cancer cells both in vitro and in vivo. METHODS： ILK small interfering RNA （siRNA） was transfected into human gastric cancer BGC-823 cells and ILK expression was monitored by real-time quan- titative polymerase chain reaction, Western blotting analysis and immunocytochemistry. Cell attachment, proliferation, invasion, microfilament dynamics and the secretion of vascular endothelial growth factor （VEGF） were also measured. Gastric cancer cells treated with ILK siRNA were subcutaneously transplanted into nude mice and tumor growth was assessed. RESULTS： Both ILK mRNA and protein levels were significantly down-regulated by ILK siRNA in human gastric cancer cells. This significantly inhibited cell attachment, proliferation and invasion. The knockdown of ILK also disturbed F-actin assembly and reduced VEGF secretion in conditioned medium by 40% （P 〈 0.05）. Four weeks after injection of ILK siRNA-transfected gastric cancer cells into nude mice, tumor volume and weight were significantly reduced compared with that of tumors induced by cells treated with non-silencing siRNA or by untreated cells （P 〈 0.05）. CONCLUSION： Targeting ILK with siRNA suppresses the growth of gastric cancer cells both in v/tro and /n vivo. ILK plays an important role in gastric cancer progression.
文摘Objective: To observe the changes of plasma concentrations of endotoxin, soluble intercellular adhesion molecule 1, tumor necrosis factor α, and urinary microalbumin in children undergoing cardiac procedure and to study the effects of cardiopulmonary bypass (CPB) on the injury or activation of endothelial cells and vascular permeability. Methods: Twenty children undergoing cardiac operation with CPB were selected in the study. Plasma concentrations of endotoxin, soluble intercellular adhesion molecule 1, tumor necrosis factor α, and urinary microalbumin were measured after anesthetic induction (baseline), bypass for 20 minutes, at the end of CPB, and at 2, 4, and 18 h after the end of CPB. Results: The plasma concentrations of endotoxin, soluble intercellular adhesion molecule 1, and urinary microalbumin began to increase at 2 h after the end of CPB, and remained higher than that of the baseline, while the concentration of tumor necrosis factor α increased only at the end CPB and at 2 h after the end of CPB. Conclusion: Cardiopulmonary bypass can induce inflammatory response, resulting in the activation or injury of vascular endothelial cells, and can increase the vascular permeability.
文摘OBJECTIVE: To investigate the reasons for the rarity of metastases in skeletal muscle. METHODS: By injecting tumor cells (Walker256 rat carcinosarcoma) through the iliac artery (experimental group) and the tail vein (control group), animal models of blood-borne metastases were established. The quadriceps femoris muscle and lungs were observed grossly and microscopically. Immunohistochemistry was applied to investigate the expression of vascular cell adhesion molecule-1 (VCAM-1) in the microvascular endothelium of these organs. Primary culture of rat skeletal muscle cells was established and conditioned medium (MCM) was collected. Effects of MCM on several tumor cell lines and the biochemical characteristics of skeletal muscle delivered tumor factor(s) were tested by MTT assay. Apoptosis and morphological examination were carried out to investigate the antitumor mechanisms of MCM. RESULTS: In the experimental group, there were no definite metastases observed in muscle cells. In the control group, lung metastases were present in the lungs of all rats that were sacrificed at the 14th day or died spontaneously (17 rats in all). There was no significant difference between the increase in VCAM-1 in quadriceps femoris muscle 7 days after iliac artery injection and that in lungs 7 days after tail vein injection (P > 0.05). In vitro studies showed that the proliferation of tumor cell lines of mouse SP2/0 myeloma, rat Walker256 carcinosarcoma or human chronic granulocytic leukemia K562, human acute lymphatic leukemia HL-60, LS-174-T colon adenocarcinoma, PC3-M prostatic carcinoma and lung giant cell carcinoma with different metastatic potency (PLA801-C with low metastatic potency, PLA801-D with high metastatic potency) was significantly inhibited when cultured with MCM (P
