AIM: To investigate the molecular mechanisms of miRNA in advanced gastric cancers (AGCs) before and after cytoreductive surgery (CRS) + hyperthermic intraperitoneal chemotherapy (HIPEC).
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〈0.0展开更多
目的:研究miR218是否通过下调SOX4影响滋养层细胞系HTR-8细胞的迁移和侵袭。方法:妊娠期高血压疾病(HDCP)患者46例,平均年龄(31±4.6)岁,收缩期血压≥140 mm Hg和/或舒张期血压>90 mm Hg;以血压正常孕妇50例为对照,实时荧光定量P...目的:研究miR218是否通过下调SOX4影响滋养层细胞系HTR-8细胞的迁移和侵袭。方法:妊娠期高血压疾病(HDCP)患者46例,平均年龄(31±4.6)岁,收缩期血压≥140 mm Hg和/或舒张期血压>90 mm Hg;以血压正常孕妇50例为对照,实时荧光定量PCR(RT-PCR)检测两组患者静脉血中miR218的表达情况。转染miR218mimic和miRNC至离体培养的HTR-8细胞中,将细胞分为对照组(加入DMEM)、空质粒组(加入miRNC)和过表达miR218组(加入miR218 mimic)3组,检测细胞的迁移侵袭情况以及细胞中MMP-2和MMP-9的表达,,生物信息学预测miR-218潜在靶基因为SOX4,利用荧光素酶素试验验证SOX4是miR218的靶基因;再通过转染过表达SOX4的质粒至HTR-8细胞,HTR-8细胞分为过表达miR218组、过表达miR218+空质粒组、过表达miR218+SOX4组,以上方法检测HTR-8细胞的迁移侵袭情况。结果:相比于正常孕妇组,HDCP组患者血清中miR218表达减少(P<0.01)。相比于空质粒组,转染miR218mimic后,HTR-8细胞中MMP-2、MMP-9、SOX4的表达减少(P<0.01),细胞迁移和侵袭能力下降(P<0.01);荧光素酶试验结果显示,miR218能够显著降低SOX4-3'-UTR质粒的荧光素活性(P<0.01);相比于miR218+空质粒组,转染过表达SOX4质粒后,HTR-8细胞迁移和侵袭能力增加(P<0.01)。结论:HDCP患者血清中miR218表达减少,miR218可以通过下调SOX4从而抑制HTR-8细胞的迁移和侵袭。展开更多
Background： The mechanisms of pathological retinal neovascularization （RNV） remain unknown. Several microRNAs were reported to be involved in the process of RNV. Oxygen-induced retinopathy （O1R） is a useful model...Background： The mechanisms of pathological retinal neovascularization （RNV） remain unknown. Several microRNAs were reported to be involved in the process of RNV. Oxygen-induced retinopathy （O1R） is a useful model to investigate RNV. Our present work explored the expression and the role of microRNA-128 （miR-218） in oxygen-induced RNV. Methods： OIR was used to establish RNV model. The expression level ofmiR-218 in the retina from OIR mice was assessed by quantitative real-time reverse transcriptase polymerase chain reaction. Fluorescein angiography was performed in retinae of OIR mice, and RNV was quantified by hematoxylin and eosin staining to evaluate the effect of pCDH-CMV-miR-218 intravitreal injection on RNV in OIR mice. Roundabout 1 （Robol） expression was detected by Western blotting in mouse retinal vascular endothelial cells expressing a high or low level of miR-218 and retinal tissues from OIR mice. Cell migration was evaluated by scratch wound assay. Results： In OIR mice, the expression level of miR-218 was significantly down-regulated （P = 0.006）. Retinal Robol expression was significantly increased at both mRNA and protein levels （P = 0.001, 0.008： respectively）, miR-218 intravitreal injection inhibited retinal angiogenesis in OIR mice, and the restoration of miR-218 in retina led to down-regulation of Robol. Conelusions： Our experiments showed that restoration ofmiR-218 inhibited retinal angiogenesis via targeting Robo 1. MiR-218 contributed to the inhibition of retinal angiogenesis and miR-218 might be a new therapeutic target for preventing RNV.展开更多
基金Supported by The PhD Start-up Funds of Guangzhou Medical College,Guangdong Province,China,No.2012C66 and No.2012C69Guangdong Province Natural Science Fund,No.S2013010016662the National Natural Science Foundation of China,No.81201932 and No.81372493
文摘AIM: To investigate the molecular mechanisms of miRNA in advanced gastric cancers (AGCs) before and after cytoreductive surgery (CRS) + hyperthermic intraperitoneal chemotherapy (HIPEC).
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〈0.0
文摘目的:研究miR218是否通过下调SOX4影响滋养层细胞系HTR-8细胞的迁移和侵袭。方法:妊娠期高血压疾病(HDCP)患者46例,平均年龄(31±4.6)岁,收缩期血压≥140 mm Hg和/或舒张期血压>90 mm Hg;以血压正常孕妇50例为对照,实时荧光定量PCR(RT-PCR)检测两组患者静脉血中miR218的表达情况。转染miR218mimic和miRNC至离体培养的HTR-8细胞中,将细胞分为对照组(加入DMEM)、空质粒组(加入miRNC)和过表达miR218组(加入miR218 mimic)3组,检测细胞的迁移侵袭情况以及细胞中MMP-2和MMP-9的表达,,生物信息学预测miR-218潜在靶基因为SOX4,利用荧光素酶素试验验证SOX4是miR218的靶基因;再通过转染过表达SOX4的质粒至HTR-8细胞,HTR-8细胞分为过表达miR218组、过表达miR218+空质粒组、过表达miR218+SOX4组,以上方法检测HTR-8细胞的迁移侵袭情况。结果:相比于正常孕妇组,HDCP组患者血清中miR218表达减少(P<0.01)。相比于空质粒组,转染miR218mimic后,HTR-8细胞中MMP-2、MMP-9、SOX4的表达减少(P<0.01),细胞迁移和侵袭能力下降(P<0.01);荧光素酶试验结果显示,miR218能够显著降低SOX4-3'-UTR质粒的荧光素活性(P<0.01);相比于miR218+空质粒组,转染过表达SOX4质粒后,HTR-8细胞迁移和侵袭能力增加(P<0.01)。结论:HDCP患者血清中miR218表达减少,miR218可以通过下调SOX4从而抑制HTR-8细胞的迁移和侵袭。
基金This study was supported by the Natural Science Foundation of Tianjin
文摘Background： The mechanisms of pathological retinal neovascularization （RNV） remain unknown. Several microRNAs were reported to be involved in the process of RNV. Oxygen-induced retinopathy （O1R） is a useful model to investigate RNV. Our present work explored the expression and the role of microRNA-128 （miR-218） in oxygen-induced RNV. Methods： OIR was used to establish RNV model. The expression level ofmiR-218 in the retina from OIR mice was assessed by quantitative real-time reverse transcriptase polymerase chain reaction. Fluorescein angiography was performed in retinae of OIR mice, and RNV was quantified by hematoxylin and eosin staining to evaluate the effect of pCDH-CMV-miR-218 intravitreal injection on RNV in OIR mice. Roundabout 1 （Robol） expression was detected by Western blotting in mouse retinal vascular endothelial cells expressing a high or low level of miR-218 and retinal tissues from OIR mice. Cell migration was evaluated by scratch wound assay. Results： In OIR mice, the expression level of miR-218 was significantly down-regulated （P = 0.006）. Retinal Robol expression was significantly increased at both mRNA and protein levels （P = 0.001, 0.008： respectively）, miR-218 intravitreal injection inhibited retinal angiogenesis in OIR mice, and the restoration of miR-218 in retina led to down-regulation of Robol. Conelusions： Our experiments showed that restoration ofmiR-218 inhibited retinal angiogenesis via targeting Robo 1. MiR-218 contributed to the inhibition of retinal angiogenesis and miR-218 might be a new therapeutic target for preventing RNV.
