AIM:To investigate the expression of microRNA-218(miR-218)in serum from gastric cancer patients and its relationship with clinicopathological characteristics.METHODS:A total of 68 patients with pathologically diagnose...AIM:To investigate the expression of microRNA-218(miR-218)in serum from gastric cancer patients and its relationship with clinicopathological characteristics.METHODS:A total of 68 patients with pathologically diagnosed gastric cancer and 56 healthy individuals were recruited to this study.The expression of miR-218was detected in the serum of gastric cancer patients and healthy individuals by quantitative real-time polymerase chain reaction.The clinical data were collectedand analyzed by statistical software.RESULTS:miR-218 was reduced significantly in the serum of gastric cancer patients compared to healthy individuals(1.15±0.08 vs 0.37±0.023;P=0.026).In the gastric cancer group,serum expression of miR-218was lower in patients with metastasis and poorly differentiated cancer compared with non-metastatic and well-differentiated cancer(0.19±0.011 vs 0.45±0.021,P=0.031 and 0.21±0.019 vs 0.49±0.021,P=0.025).Serum miR-218 was found to be significantly associated with gastric cancer metastasis(P=0.003),tumor T stage(P=0.018)and tumor grade(P=0.012).Low serum expression of miR-218 was related to an increase in the stage of gastric cancer.The expression level of miR-218 in the serum was correlated with the3-year survival.Ninety-seven percent of patients with a high level of miR-218 expression survived for 3 years,while only 54%of those with low miR-218 expression survived.CONCLUSION:miR-218 is deregulated in gastric cancer patients and is strongly correlated with tumor stage,grade and metastasis.Serum expression of miR-218 may be a prognostic marker.展开更多
Objective： To detect the expression profiles of micro RNA-218（mi R-218） in human pancreatic cancer tissue（PCT） and cells and their effects on the biological features of human pancreatic cancer cell line PANC-1 an...Objective： To detect the expression profiles of micro RNA-218（mi R-218） in human pancreatic cancer tissue（PCT） and cells and their effects on the biological features of human pancreatic cancer cell line PANC-1 and observe the effect of mi R-218 on the expression of the target gene high mobility group box 1（HMGB1）, with an attempt to provide new treatment methods and strategies for pancreatic cancer.Methods： The expressions of mi R-218 in PCT and normal pancreas tissue as well as in various pancreatic cancer cell lines including As PC-1, Bx PC-3, and PANC-1 were determined with quantitative real-time reverse transcription polymerase chain reaction（q RT-PCR）. The change of mi R-218 expression in PANC-1 cells was detected using qR T-PCT after the transfection of miR-218 mimic for 48 h. Cell Counting Kit-8（CCK-8） was applied for detecting the effect of mi R-218 on the activity of PANC-1 cells. The effects of mi R-218 on the proliferation and apoptosis of PANC-1 cells were analyzed using the flow cytometry. The effect of mi R-218 on the migration of PANC-1 cells was detected using the Trans-well migration assay. The HMGB1 was found to be a target gene of mi R-218 by luciferase reporter assay, and the effect of mi R-218 on the expression of HMGB1 protein in cells were determined using Western blotting.Results： As shown by q RT-PCR, the expressions of mi R-218 in PCT and in pancreatic cancer cell line significantly decreased when compared with the normal pancreatic tissue（NPT）（P〈0.01）. Compared with the control group, the miR-218 expression significantly increased in the PANC-1 group after the transfection of mi R-218 mimic for 48 h（P〈0.01）. Growth curve showed that the cell viability significantly dropped after the overexpression of mi R-218 in the PANC-1 cells for two days（P〈0.05）. Flow cytometry showed that the S-phase fraction significantly dropped after the overexpression of mi R-218（P〈0.01） and the percentage of apoptotic cells significantly increased（P〈0.01）. As shown by the Trans-well migration assay, the enhanced mi R-218 expression was associated with a significantly lower number of cells that passed through a Transwell chamber（P〈0.01）. Luciferase reporter assay showed that, compared with the control group, the relative luciferase activity significantly decreased in the mi R-218 mimic group（P〈0.01）. As shown by the Western blotting, compared with the control group, the HMGB1 protein expression significantly decreased in the PANC-1 group after the transfection of mi R-218 mimic for 48 h（P〈0.01）.Conclusions： The mi R-218 expression decreases in human PCT and cell lines. mi R-218 can negatively regulate the HMGB1 protein expression and inhibit the proliferation and invasion of pancreatic cancer cells. A treatment strategy by enhancing the mi R-218 expression may benefit the patients with pancreatic cancer.展开更多
Objective： The purpose of this study was to examine the effect of gemcitabine（GEM） on micro RNA-218（mi R-218） expression in human pancreatic cancer cells.Methods： Quantitative reverse transcription polymerase ch...Objective： The purpose of this study was to examine the effect of gemcitabine（GEM） on micro RNA-218（mi R-218） expression in human pancreatic cancer cells.Methods： Quantitative reverse transcription polymerase chain reaction（q RT-PCR） was performed to examine the differences in mi R-218 expression between the GEM-sensitive Bx PC-3 pancreatic cancer cells and GEM-resistant PANC-1 cells. The effect of GEM on the expression of mi R-218 in PANC-1 cells was also investigated. PANC-1 cells were transfected either with HMGB1 si RNA to knock down the expression of HMGB1 or with the recombinant HMGB1 expression vector（pc DNA3.1-HMGB1） to overexpress HMGB1. The effect of ectopic expression of HMGB1 on the apoptosis of mi R-218-transfected and GEMtreated PANC-1 cells was examined by flow cytometric analysis.Results： The mi R-218 expression level was lower in GEM-resistant PANC-1 cells compared to GEMsensitive Bx PC-3 cells（P〈0.05）. The percentage of apoptotic PANC-1 cells was significantly increased in the mi R-218 mimic ＋ GEM group compared to the mimic ctrl ＋ GEM group and the normal control group（P〈0.01）. The HMGB1 expression level was markedly decreased in PANC-1 cells transfected with HMGB1 si RNA but was significantly increased in PANC-1 cells transfected with the recombinant HMGB1 expression vector, pc DNA3.1-HMGB1（P〈0.01）. The proportion of apoptotic PANC-1 cells was significantly lower in the mi R-218 mimic ＋ GEM ＋ pc DNA3.1-HMGB1 group compared to the mi R-218 mimic ＋ GEM ＋ HMGB1 si RNA group（P〈0.01）.Conclusions： The expression level of mi R-218 was downregulated in the GEM-resistant cell line. mi R-218 promoted the sensitivity of PANC-1 cells to GEM, which was achieved mainly through regulating the expression of HMGB1 in PANC-1 cells.展开更多
基金Supported by A grant from Liaoning Provincial Science and Technology Plan Foundation,No.2011225013
文摘AIM:To investigate the expression of microRNA-218(miR-218)in serum from gastric cancer patients and its relationship with clinicopathological characteristics.METHODS:A total of 68 patients with pathologically diagnosed gastric cancer and 56 healthy individuals were recruited to this study.The expression of miR-218was detected in the serum of gastric cancer patients and healthy individuals by quantitative real-time polymerase chain reaction.The clinical data were collectedand analyzed by statistical software.RESULTS:miR-218 was reduced significantly in the serum of gastric cancer patients compared to healthy individuals(1.15±0.08 vs 0.37±0.023;P=0.026).In the gastric cancer group,serum expression of miR-218was lower in patients with metastasis and poorly differentiated cancer compared with non-metastatic and well-differentiated cancer(0.19±0.011 vs 0.45±0.021,P=0.031 and 0.21±0.019 vs 0.49±0.021,P=0.025).Serum miR-218 was found to be significantly associated with gastric cancer metastasis(P=0.003),tumor T stage(P=0.018)and tumor grade(P=0.012).Low serum expression of miR-218 was related to an increase in the stage of gastric cancer.The expression level of miR-218 in the serum was correlated with the3-year survival.Ninety-seven percent of patients with a high level of miR-218 expression survived for 3 years,while only 54%of those with low miR-218 expression survived.CONCLUSION:miR-218 is deregulated in gastric cancer patients and is strongly correlated with tumor stage,grade and metastasis.Serum expression of miR-218 may be a prognostic marker.
基金supported by grants:Liaoning Provincial Department of education science research project(L2014299)Liaoning province science and technology plan project(2011404013-4)The Shenyang Municipal Science and technology project(F12-277-1-73)
文摘Objective： To detect the expression profiles of micro RNA-218（mi R-218） in human pancreatic cancer tissue（PCT） and cells and their effects on the biological features of human pancreatic cancer cell line PANC-1 and observe the effect of mi R-218 on the expression of the target gene high mobility group box 1（HMGB1）, with an attempt to provide new treatment methods and strategies for pancreatic cancer.Methods： The expressions of mi R-218 in PCT and normal pancreas tissue as well as in various pancreatic cancer cell lines including As PC-1, Bx PC-3, and PANC-1 were determined with quantitative real-time reverse transcription polymerase chain reaction（q RT-PCR）. The change of mi R-218 expression in PANC-1 cells was detected using qR T-PCT after the transfection of miR-218 mimic for 48 h. Cell Counting Kit-8（CCK-8） was applied for detecting the effect of mi R-218 on the activity of PANC-1 cells. The effects of mi R-218 on the proliferation and apoptosis of PANC-1 cells were analyzed using the flow cytometry. The effect of mi R-218 on the migration of PANC-1 cells was detected using the Trans-well migration assay. The HMGB1 was found to be a target gene of mi R-218 by luciferase reporter assay, and the effect of mi R-218 on the expression of HMGB1 protein in cells were determined using Western blotting.Results： As shown by q RT-PCR, the expressions of mi R-218 in PCT and in pancreatic cancer cell line significantly decreased when compared with the normal pancreatic tissue（NPT）（P〈0.01）. Compared with the control group, the miR-218 expression significantly increased in the PANC-1 group after the transfection of mi R-218 mimic for 48 h（P〈0.01）. Growth curve showed that the cell viability significantly dropped after the overexpression of mi R-218 in the PANC-1 cells for two days（P〈0.05）. Flow cytometry showed that the S-phase fraction significantly dropped after the overexpression of mi R-218（P〈0.01） and the percentage of apoptotic cells significantly increased（P〈0.01）. As shown by the Trans-well migration assay, the enhanced mi R-218 expression was associated with a significantly lower number of cells that passed through a Transwell chamber（P〈0.01）. Luciferase reporter assay showed that, compared with the control group, the relative luciferase activity significantly decreased in the mi R-218 mimic group（P〈0.01）. As shown by the Western blotting, compared with the control group, the HMGB1 protein expression significantly decreased in the PANC-1 group after the transfection of mi R-218 mimic for 48 h（P〈0.01）.Conclusions： The mi R-218 expression decreases in human PCT and cell lines. mi R-218 can negatively regulate the HMGB1 protein expression and inhibit the proliferation and invasion of pancreatic cancer cells. A treatment strategy by enhancing the mi R-218 expression may benefit the patients with pancreatic cancer.
基金supported by grants:Liaoning Provincial Department of Education Science Research Project(L2014299)Liaoning Province Science and Technology Plan Project(2011404013-4)The Shenyang Municipal Science and Technology Project(F12-277-1-73)
文摘Objective： The purpose of this study was to examine the effect of gemcitabine（GEM） on micro RNA-218（mi R-218） expression in human pancreatic cancer cells.Methods： Quantitative reverse transcription polymerase chain reaction（q RT-PCR） was performed to examine the differences in mi R-218 expression between the GEM-sensitive Bx PC-3 pancreatic cancer cells and GEM-resistant PANC-1 cells. The effect of GEM on the expression of mi R-218 in PANC-1 cells was also investigated. PANC-1 cells were transfected either with HMGB1 si RNA to knock down the expression of HMGB1 or with the recombinant HMGB1 expression vector（pc DNA3.1-HMGB1） to overexpress HMGB1. The effect of ectopic expression of HMGB1 on the apoptosis of mi R-218-transfected and GEMtreated PANC-1 cells was examined by flow cytometric analysis.Results： The mi R-218 expression level was lower in GEM-resistant PANC-1 cells compared to GEMsensitive Bx PC-3 cells（P〈0.05）. The percentage of apoptotic PANC-1 cells was significantly increased in the mi R-218 mimic ＋ GEM group compared to the mimic ctrl ＋ GEM group and the normal control group（P〈0.01）. The HMGB1 expression level was markedly decreased in PANC-1 cells transfected with HMGB1 si RNA but was significantly increased in PANC-1 cells transfected with the recombinant HMGB1 expression vector, pc DNA3.1-HMGB1（P〈0.01）. The proportion of apoptotic PANC-1 cells was significantly lower in the mi R-218 mimic ＋ GEM ＋ pc DNA3.1-HMGB1 group compared to the mi R-218 mimic ＋ GEM ＋ HMGB1 si RNA group（P〈0.01）.Conclusions： The expression level of mi R-218 was downregulated in the GEM-resistant cell line. mi R-218 promoted the sensitivity of PANC-1 cells to GEM, which was achieved mainly through regulating the expression of HMGB1 in PANC-1 cells.
