AIM: To determine the expression of micro RNA-210(mi R-210) in hepatocellular carcinoma(HCC) and to examine its role using HCC cells.METHODS: The expression of mi R-210 was determined in 21 pairs of HCC samples and th...AIM: To determine the expression of micro RNA-210(mi R-210) in hepatocellular carcinoma(HCC) and to examine its role using HCC cells.METHODS: The expression of mi R-210 was determined in 21 pairs of HCC samples and the corresponding surrounding non-tumor tissues. The effects of mi R-210 on proliferation and cell cycle progression were examined using Hep G2 and Hu H7 cells. Overexpression and inhibition of mi R-210 was achieved by transfection of the cells with mi R-210 mimic or inhibitor. Luciferase reporter constructs were used to identify the mi R-210 interacting site on Yes1. Yes1 expression was examined after mi R-210 transfection,as well as in the HCC samples.RESULTS: mi R-210 was significantly up-regulated by 3.4 fold(P < 0.01) in the tumor samples. The over-expression of mi R-210 significantly reduced cell proliferation compared to the mock-treated cells(68.9% ± 7.4% and 53.6% ± 5.0%,P < 0.05 for the Hep G2 and Hu H7 cells respectively). Analysis of the Hu H7 cells transfected with mi R-210 mimic by flow cytometry showed that the cells took a longer time to reach the G2/M phase. The interaction between mi R-210 and the 3'UTR of the Yes1 transcript was confirmed using a luciferase reporter assay. Over-expression of mi R-210 reduced the expression of Yes1 protein in both Hu H7 and Hep G2 cells. Tumors with a greater than fourfold increase in the expression of mi R-210 showed consistently lower expressions of Yes1 in the tumors.In nocodazole-treated cells with a significant G2/M cell population,Yes1 protein was significantly reduced and pre-inhibition of mi R-210 in Hu H7 cells was able to prevent the reduction of Yes1 protein expression. Knock-down of Yes1 by si RNA also led to reduced cell proliferation(70.8% ± 7.5%,P < 0.05 in the Hu H7 cells).CONCLUSION: Up-regulation of mi R-210 inhibits cell proliferation. Yes1 is a target of mi R-210 and affects cell proliferation in HCC.展开更多
Objective： To investigate the microR NAs（miR NAs） expression profile of acute myocardial infarction（AMI） rats and the regulating effects of Huoxue Anxin Recipe（活血安心方, HAR） on angiogenesis-related miR NAs a...Objective： To investigate the microR NAs（miR NAs） expression profile of acute myocardial infarction（AMI） rats and the regulating effects of Huoxue Anxin Recipe（活血安心方, HAR） on angiogenesis-related miR NAs and genes. Methods： Forty-five Wistar rats were randomly assigned to 3 groups according to a random number table： sham, AMI, and AMI＋HAR groups（15 in each group）. AMI rats were established by ligation of the left descending coronary artery. HAR was intragastrically administered to rats of the AMI＋HAR group for successive 21 days since modeling, meanwhile the same volume of 0.9% normal saline was administered to rats of the sham and AMI groups. Doppler echocardiography was used for noninvasive cardiac function test. Hematoxylin and eosin staining was used to observe the histopathological change. miR NAs expression profile was detected by quantitative realtime polymerase chain reaction（qR T-PCR）. The mR NA and protein expressions of vascular endothelial growth factor（VEGF）, and a target gene of miR-210 was further detected by qR T-PCR and Western blot, respectively. The microvessels density of myocardium was evaluated by CD31 immunostaining. Results： Compared with the sham group, ejection fraction（EF） and fractional shortening（FS） values were decreased significantly in the AMI group（P〈0.01）, while the infarction area and the interstitial collagen deposition were increased obviously. As for the AMI＋HAR group, EF and FS values were increased significantly（P〈0.05 vs. AMI group）, and the infarction area was reduced and the interstitial collagen deposition were alleviated significantly. Total of 23 miR NAs in the AMI group expressed differently by at least 1.5 folds compared with those in the sham group; 5 miR NAs in the AMI＋HAR group expressed differently by at least 1.5 folds compared with those in the AMI group. Among them, miR-210 was low in the AMI group and high in the AMI＋HAR group. The relative mR NA and protein expressions of VEGF were decreased significantly in the AMI group（P〈0.05 vs. sham group）, and increased significantly in the AMI＋HAR group（P〈0.01 vs. AMI group）. CD31 expression area and optical intensity were decreased significantly in the AMI group（P〈0.05 vs. sham group）, and increased significantly in the AMI＋HAR group（P〈0.01 vs. AMI group）. Conclusions： HAR could reduce the infarction area, alleviate the interstitial fibrosis and improve the cardiac function of AMI rats. Those effects could be related to promoting myocardium angiogenesis of HAR by up-regulating miR-210 and VEGF.展开更多
基金Supported by Biomedical Research Council and Ministry of Education(Tier 1)awarded to Tan TM
文摘AIM: To determine the expression of micro RNA-210(mi R-210) in hepatocellular carcinoma(HCC) and to examine its role using HCC cells.METHODS: The expression of mi R-210 was determined in 21 pairs of HCC samples and the corresponding surrounding non-tumor tissues. The effects of mi R-210 on proliferation and cell cycle progression were examined using Hep G2 and Hu H7 cells. Overexpression and inhibition of mi R-210 was achieved by transfection of the cells with mi R-210 mimic or inhibitor. Luciferase reporter constructs were used to identify the mi R-210 interacting site on Yes1. Yes1 expression was examined after mi R-210 transfection,as well as in the HCC samples.RESULTS: mi R-210 was significantly up-regulated by 3.4 fold(P < 0.01) in the tumor samples. The over-expression of mi R-210 significantly reduced cell proliferation compared to the mock-treated cells(68.9% ± 7.4% and 53.6% ± 5.0%,P < 0.05 for the Hep G2 and Hu H7 cells respectively). Analysis of the Hu H7 cells transfected with mi R-210 mimic by flow cytometry showed that the cells took a longer time to reach the G2/M phase. The interaction between mi R-210 and the 3'UTR of the Yes1 transcript was confirmed using a luciferase reporter assay. Over-expression of mi R-210 reduced the expression of Yes1 protein in both Hu H7 and Hep G2 cells. Tumors with a greater than fourfold increase in the expression of mi R-210 showed consistently lower expressions of Yes1 in the tumors.In nocodazole-treated cells with a significant G2/M cell population,Yes1 protein was significantly reduced and pre-inhibition of mi R-210 in Hu H7 cells was able to prevent the reduction of Yes1 protein expression. Knock-down of Yes1 by si RNA also led to reduced cell proliferation(70.8% ± 7.5%,P < 0.05 in the Hu H7 cells).CONCLUSION: Up-regulation of mi R-210 inhibits cell proliferation. Yes1 is a target of mi R-210 and affects cell proliferation in HCC.
基金Supported by the Major New Drug Development Program from the Ministry of Science and Technology,China(No.2012ZX09102-201-006)
文摘Objective： To investigate the microR NAs（miR NAs） expression profile of acute myocardial infarction（AMI） rats and the regulating effects of Huoxue Anxin Recipe（活血安心方, HAR） on angiogenesis-related miR NAs and genes. Methods： Forty-five Wistar rats were randomly assigned to 3 groups according to a random number table： sham, AMI, and AMI＋HAR groups（15 in each group）. AMI rats were established by ligation of the left descending coronary artery. HAR was intragastrically administered to rats of the AMI＋HAR group for successive 21 days since modeling, meanwhile the same volume of 0.9% normal saline was administered to rats of the sham and AMI groups. Doppler echocardiography was used for noninvasive cardiac function test. Hematoxylin and eosin staining was used to observe the histopathological change. miR NAs expression profile was detected by quantitative realtime polymerase chain reaction（qR T-PCR）. The mR NA and protein expressions of vascular endothelial growth factor（VEGF）, and a target gene of miR-210 was further detected by qR T-PCR and Western blot, respectively. The microvessels density of myocardium was evaluated by CD31 immunostaining. Results： Compared with the sham group, ejection fraction（EF） and fractional shortening（FS） values were decreased significantly in the AMI group（P〈0.01）, while the infarction area and the interstitial collagen deposition were increased obviously. As for the AMI＋HAR group, EF and FS values were increased significantly（P〈0.05 vs. AMI group）, and the infarction area was reduced and the interstitial collagen deposition were alleviated significantly. Total of 23 miR NAs in the AMI group expressed differently by at least 1.5 folds compared with those in the sham group; 5 miR NAs in the AMI＋HAR group expressed differently by at least 1.5 folds compared with those in the AMI group. Among them, miR-210 was low in the AMI group and high in the AMI＋HAR group. The relative mR NA and protein expressions of VEGF were decreased significantly in the AMI group（P〈0.05 vs. sham group）, and increased significantly in the AMI＋HAR group（P〈0.01 vs. AMI group）. CD31 expression area and optical intensity were decreased significantly in the AMI group（P〈0.05 vs. sham group）, and increased significantly in the AMI＋HAR group（P〈0.01 vs. AMI group）. Conclusions： HAR could reduce the infarction area, alleviate the interstitial fibrosis and improve the cardiac function of AMI rats. Those effects could be related to promoting myocardium angiogenesis of HAR by up-regulating miR-210 and VEGF.
