AIM:To reveal the functions of micro RNAs(mi RNAs) with respect to hepatic stellate cells(HSCs) in response to portal hypertension.METHODS:Primary rat HSCs were exposed to static water pressure(10 mm Hg,1 h) and the p...AIM:To reveal the functions of micro RNAs(mi RNAs) with respect to hepatic stellate cells(HSCs) in response to portal hypertension.METHODS:Primary rat HSCs were exposed to static water pressure(10 mm Hg,1 h) and the pressureinduced mi RNA expression profile was detected by next-generation sequencing. Quantitative real-time polymerase chain reaction was used to verify the expression of mi RNAs. A potential target of Mi R-9a-5p was measured by a luciferase reporter assay and Western blot. CCK-8 assay and Transwell assay were used to detect the proliferation and migration of HSCs under pressure.RESULTS:According to the profile,the expression of mi R-9a-5p was further confirmed to be significantly increased after pressure overload in HSCs(3.70 ± 0.61 vs 0.97 ± 0.15,P = 0.0226),which resulted in the proliferation,migration and activation of HSCs. In vivo,the up-regulation of mi R-9a-5p(2.09 ± 0.91 vs 4.27 ± 1.74,P = 0.0025) and the down-regulation of Sirt1(2.41 ± 0.51 vs 1.13 ± 0.11,P = 0.0006) were observed in rat fibrotic liver with portal hypertension. Sirt1 was a potential target gene of mi R-9a-5p. Through restoringthe expression of Sirt1 in mi R-9a-5p transfected HSCs on pressure overload,we found that overexpression of Sirt1 could partially abrogate the mi R-9a-5p mediated suppression of the proliferation,migration and activation of HSCs. CONCLUSION:Our results suggest that during liver fibrosis,portal hypertension may induce the proliferation,migration and activation of HSCs through the up-regulation of mi R-9a-5p,which targets Sirt1.展开更多
AIM: To elucidate the potential biological role of mi R-30 b in gastric cancer and investigate the underlying molecular mechanisms of mi R-30 b to inhibit metastasis of gastric cancer cells.METHODS: The expression of ...AIM: To elucidate the potential biological role of mi R-30 b in gastric cancer and investigate the underlying molecular mechanisms of mi R-30 b to inhibit metastasis of gastric cancer cells.METHODS: The expression of mi R-30 b was detected in gastric cancer cell lines and samples by reverse transcription-polymerase chain reaction. CCK-8 assays were conducted to explore the impact of mi R-30 b overexpression on the proliferation of gastric cancer cells. Flow cytometry was used to examine the effect of mi R-30 b on the apoptosis. Transwell test was used for the migration and invasion assays. Luciferase reporter assays and Western blot were employed to validate regulation of putative target of mi R-30 b.RESULTS: The results showed that mi R-30 b was downregulated in gastric cancer tissues and cancer cell lines and functioned as a tumor suppressor. Overexpression of mi R-30 b promoted cell apoptosis,and suppressed proliferation,migration and invasion of the gastric cancer cell lines AGS and MGC803. Bioinformatic analysis identified the 3'-untranslated region of eukaryotic translation initiation factor 5A2(EIF5A2) as a putative binding site of mi R-30 b. Luciferase reporter assays and Western blot analysis confirmed the EIF5A2 gene as a target of mi R-30 b. Moreover,expression levels of theEIF5A2 targets E-cadherin and Vimentin were altered following transfection of mi R-30 b mimics.CONCLUSION: Our findings describe a link between mi R-30 b and EIF5A2,which plays an important role in mediating epithelial-mesenchymal transition.展开更多
AIM To evaluate the levels of mi R-192-5 p in non-alcoholic fatty liver disease(NAFLD) models and demonstrate the role of mi R-192-5 p in lipid accumulation. METHODS Thirty Sprague Dawley rats were randomly divided in...AIM To evaluate the levels of mi R-192-5 p in non-alcoholic fatty liver disease(NAFLD) models and demonstrate the role of mi R-192-5 p in lipid accumulation. METHODS Thirty Sprague Dawley rats were randomly divided into three groups, which were given a standard diet, a high-fat diet(HFD), and an HFD with injection of liraglutide. At the end of 16 weeks, hepatic mi R-192-5 p and stearoyl-Co A desaturase 1(SCD-1) levels were measured. Mi R-192-5 p mimic and inhibitor and SCD-1 si RNA were transfected into Huh7 cells exposed to palmitic acid(PA). Lipid accumulation was evaluated by oil red O staining and triglyceride assays. Direct interaction was validated by dual-luciferase reporter gene assays.RESULTS The HFD rats showed a 0.46-fold decrease and a 3.5-fold increase in hepatic mi R-192-5 p and SCD-1 protein levels compared with controls, respectively, which could be reversed after disease remission by liraglutide injection(P < 0.01). The Huh7 cells exposed to PA also showed down-regulation and up-regulation of mi R-192-5 p and SCD-1 protein levels, respectively(P < 0.01). Transfection with mi R-192-5 p mimic and inhibitor in Huh7 cells induced dramatic repression and promotion of SCD-1 protein levels, respectively(P < 0.01). Luciferase activity was suppressed and enhanced by mi R-192-5 p mimic and inhibitor, respectively, in wild-type SCD-1(P < 0.01) but not in mutant SCD-1. Mi R-192-5 p overexpression reduced lipid accumulation significantly in PA-treated Huh7 cells, and SCD-1 si RNA transfection abrogated the lipid deposition aggravated by mi R-192-5 p inhibitor(P < 0.01).CONCLUSION This study demonstrates that mi R-192-5 p has a negative regulatory role in lipid synthesis, which is mediated through its direct regulation of SCD-1.展开更多
Micro RNAs are small non-coding RNAs that participate in different biological processes, providing subtle combinational regulation of cellular pathways, often by regulating components of signalling pathways. Aberrant ...Micro RNAs are small non-coding RNAs that participate in different biological processes, providing subtle combinational regulation of cellular pathways, often by regulating components of signalling pathways. Aberrant expression of mi RNAs is an important factor in the development and progression of disease. The canonical myomi Rs(mi R-1,-133 and-206) are central to the development and health of mammalian skeletal and cardiac muscles, but new findings show they have regulatory roles in the development of other mammalian non-muscle tissues, including nerve, brain structures, adipose and some specialised immunological cells. Moreover, the deregulation of myomi R expression is associated with a variety of different cancers, where typically they have tumor suppressor functions, although examples of an oncogenic role illustrate their diverse function in different cell environments. This review examines the involvement of the related myomi Rs at the crossroads between cell development/tissue regeneration/tissue inflammation responses, and cancer development.展开更多
A mango ETHYLENE RESPONSE1(ETR1) gene, designated Mi ETR1 b, was isolated from the cotyledon of mango(Mangifera indica L. ‘Zihua')using RT-PCR, and the 5′ and 3′ rapid amplificatio of c DNA ends. The full-lengt...A mango ETHYLENE RESPONSE1(ETR1) gene, designated Mi ETR1 b, was isolated from the cotyledon of mango(Mangifera indica L. ‘Zihua')using RT-PCR, and the 5′ and 3′ rapid amplificatio of c DNA ends. The full-length c DNA was 2 530 bp, with an open reading frame of 2 220 bp,and it encoded a putative protein of 739 amino acids. The genomic DNA sequence of Mi ETR1 b was 4 116 bp in length, having a 3 305 bp sequence from the start to terminator codon, containing six exons and fi e introns. The deduced amino acids possessed conserved domains of the GAF and HATPase_c superfamilies. A phylogenetic tree analysis indicated that Mi ETR1 b had the highest similarity to Mi ETR1 from M. indica and a high similarity to Cs ERS1, Dl ETR1, Tc ERS1 and Pt ETR1. Quantitative real-time PCR showed that Mi ETR1 b was expressed in the proximal and distal cut surfaces throughout the adventitious root formation period. Meanwhile, the expression of Mi ETR1 b in the distal cut surface was significant y up-regulated within 6–48 h. Pre-treatments with indole-3-butyric acid and 2,3,5-triiodobenzoic acid significant y downregulated Mi ETR1 b expression at 1 d and 6 h, respectively. However, more ethylene was produced from 12 to 24 h, while ethylene production decreased after 4 days of culturing. In conclusion, Mi ETR1 b might play an important role during the adventitious root formation of mango cotyledon segments, which is related to ethylene production.展开更多
目的探讨过表达mi R-340对人肝癌Hep G2、MHCC-97L细胞增殖、细胞周期和凋亡的影响。方法 mi R-340 mimic转染后RT-PCR法测定肝癌细胞株mi R-340 m RNA水平,Brdu-ELISA试剂盒检测细胞增殖,流式细胞术分析细胞周期,膜联蛋白V-FITC凋亡检...目的探讨过表达mi R-340对人肝癌Hep G2、MHCC-97L细胞增殖、细胞周期和凋亡的影响。方法 mi R-340 mimic转染后RT-PCR法测定肝癌细胞株mi R-340 m RNA水平,Brdu-ELISA试剂盒检测细胞增殖,流式细胞术分析细胞周期,膜联蛋白V-FITC凋亡检测试剂盒分析细胞凋亡。结果mi R-340在肝癌细胞系中表达降低(P<0.05);高表达mi R-340后肝癌细胞的存活率受到显著抑制(P<0.05);G0/G1期细胞增多(P<0.05);S期细胞减少(P<0.05);高表达mi R-340后Hep G2和MHCC-97L细胞凋亡受到促进(P<0.05)。结论 mi R-340在抑制细胞增殖、NF-κB1下调诱导的细胞凋亡中起着重要的作用,提示mi R-340表达降低是肝细胞肝癌发生的机制之一。展开更多
基金Supported by National Natural Science Foundation of China,No.11272342/A0205
文摘AIM:To reveal the functions of micro RNAs(mi RNAs) with respect to hepatic stellate cells(HSCs) in response to portal hypertension.METHODS:Primary rat HSCs were exposed to static water pressure(10 mm Hg,1 h) and the pressureinduced mi RNA expression profile was detected by next-generation sequencing. Quantitative real-time polymerase chain reaction was used to verify the expression of mi RNAs. A potential target of Mi R-9a-5p was measured by a luciferase reporter assay and Western blot. CCK-8 assay and Transwell assay were used to detect the proliferation and migration of HSCs under pressure.RESULTS:According to the profile,the expression of mi R-9a-5p was further confirmed to be significantly increased after pressure overload in HSCs(3.70 ± 0.61 vs 0.97 ± 0.15,P = 0.0226),which resulted in the proliferation,migration and activation of HSCs. In vivo,the up-regulation of mi R-9a-5p(2.09 ± 0.91 vs 4.27 ± 1.74,P = 0.0025) and the down-regulation of Sirt1(2.41 ± 0.51 vs 1.13 ± 0.11,P = 0.0006) were observed in rat fibrotic liver with portal hypertension. Sirt1 was a potential target gene of mi R-9a-5p. Through restoringthe expression of Sirt1 in mi R-9a-5p transfected HSCs on pressure overload,we found that overexpression of Sirt1 could partially abrogate the mi R-9a-5p mediated suppression of the proliferation,migration and activation of HSCs. CONCLUSION:Our results suggest that during liver fibrosis,portal hypertension may induce the proliferation,migration and activation of HSCs through the up-regulation of mi R-9a-5p,which targets Sirt1.
基金Supported by Beijing Municipal Natural Science Foundation of China,No.7132209The Capital Health Research and Development of Special,No.2014-3-4014
文摘AIM: To elucidate the potential biological role of mi R-30 b in gastric cancer and investigate the underlying molecular mechanisms of mi R-30 b to inhibit metastasis of gastric cancer cells.METHODS: The expression of mi R-30 b was detected in gastric cancer cell lines and samples by reverse transcription-polymerase chain reaction. CCK-8 assays were conducted to explore the impact of mi R-30 b overexpression on the proliferation of gastric cancer cells. Flow cytometry was used to examine the effect of mi R-30 b on the apoptosis. Transwell test was used for the migration and invasion assays. Luciferase reporter assays and Western blot were employed to validate regulation of putative target of mi R-30 b.RESULTS: The results showed that mi R-30 b was downregulated in gastric cancer tissues and cancer cell lines and functioned as a tumor suppressor. Overexpression of mi R-30 b promoted cell apoptosis,and suppressed proliferation,migration and invasion of the gastric cancer cell lines AGS and MGC803. Bioinformatic analysis identified the 3'-untranslated region of eukaryotic translation initiation factor 5A2(EIF5A2) as a putative binding site of mi R-30 b. Luciferase reporter assays and Western blot analysis confirmed the EIF5A2 gene as a target of mi R-30 b. Moreover,expression levels of theEIF5A2 targets E-cadherin and Vimentin were altered following transfection of mi R-30 b mimics.CONCLUSION: Our findings describe a link between mi R-30 b and EIF5A2,which plays an important role in mediating epithelial-mesenchymal transition.
基金Supported by National Key R&D Program of China No.2017YFC0908900National Key Basic Research Project,No.2012CB517501National Natural Science Foundation of China,No.81470840 and No.81600464
文摘AIM To evaluate the levels of mi R-192-5 p in non-alcoholic fatty liver disease(NAFLD) models and demonstrate the role of mi R-192-5 p in lipid accumulation. METHODS Thirty Sprague Dawley rats were randomly divided into three groups, which were given a standard diet, a high-fat diet(HFD), and an HFD with injection of liraglutide. At the end of 16 weeks, hepatic mi R-192-5 p and stearoyl-Co A desaturase 1(SCD-1) levels were measured. Mi R-192-5 p mimic and inhibitor and SCD-1 si RNA were transfected into Huh7 cells exposed to palmitic acid(PA). Lipid accumulation was evaluated by oil red O staining and triglyceride assays. Direct interaction was validated by dual-luciferase reporter gene assays.RESULTS The HFD rats showed a 0.46-fold decrease and a 3.5-fold increase in hepatic mi R-192-5 p and SCD-1 protein levels compared with controls, respectively, which could be reversed after disease remission by liraglutide injection(P < 0.01). The Huh7 cells exposed to PA also showed down-regulation and up-regulation of mi R-192-5 p and SCD-1 protein levels, respectively(P < 0.01). Transfection with mi R-192-5 p mimic and inhibitor in Huh7 cells induced dramatic repression and promotion of SCD-1 protein levels, respectively(P < 0.01). Luciferase activity was suppressed and enhanced by mi R-192-5 p mimic and inhibitor, respectively, in wild-type SCD-1(P < 0.01) but not in mutant SCD-1. Mi R-192-5 p overexpression reduced lipid accumulation significantly in PA-treated Huh7 cells, and SCD-1 si RNA transfection abrogated the lipid deposition aggravated by mi R-192-5 p inhibitor(P < 0.01).CONCLUSION This study demonstrates that mi R-192-5 p has a negative regulatory role in lipid synthesis, which is mediated through its direct regulation of SCD-1.
基金Supported by National High-tech Program of China,Nos.2006AA020701 and 2009AA022701
文摘Micro RNAs are small non-coding RNAs that participate in different biological processes, providing subtle combinational regulation of cellular pathways, often by regulating components of signalling pathways. Aberrant expression of mi RNAs is an important factor in the development and progression of disease. The canonical myomi Rs(mi R-1,-133 and-206) are central to the development and health of mammalian skeletal and cardiac muscles, but new findings show they have regulatory roles in the development of other mammalian non-muscle tissues, including nerve, brain structures, adipose and some specialised immunological cells. Moreover, the deregulation of myomi R expression is associated with a variety of different cancers, where typically they have tumor suppressor functions, although examples of an oncogenic role illustrate their diverse function in different cell environments. This review examines the involvement of the related myomi Rs at the crossroads between cell development/tissue regeneration/tissue inflammation responses, and cancer development.
基金financial y supported by the National Natural Science Foundation of China(31372053)
文摘A mango ETHYLENE RESPONSE1(ETR1) gene, designated Mi ETR1 b, was isolated from the cotyledon of mango(Mangifera indica L. ‘Zihua')using RT-PCR, and the 5′ and 3′ rapid amplificatio of c DNA ends. The full-length c DNA was 2 530 bp, with an open reading frame of 2 220 bp,and it encoded a putative protein of 739 amino acids. The genomic DNA sequence of Mi ETR1 b was 4 116 bp in length, having a 3 305 bp sequence from the start to terminator codon, containing six exons and fi e introns. The deduced amino acids possessed conserved domains of the GAF and HATPase_c superfamilies. A phylogenetic tree analysis indicated that Mi ETR1 b had the highest similarity to Mi ETR1 from M. indica and a high similarity to Cs ERS1, Dl ETR1, Tc ERS1 and Pt ETR1. Quantitative real-time PCR showed that Mi ETR1 b was expressed in the proximal and distal cut surfaces throughout the adventitious root formation period. Meanwhile, the expression of Mi ETR1 b in the distal cut surface was significant y up-regulated within 6–48 h. Pre-treatments with indole-3-butyric acid and 2,3,5-triiodobenzoic acid significant y downregulated Mi ETR1 b expression at 1 d and 6 h, respectively. However, more ethylene was produced from 12 to 24 h, while ethylene production decreased after 4 days of culturing. In conclusion, Mi ETR1 b might play an important role during the adventitious root formation of mango cotyledon segments, which is related to ethylene production.
