Objective Resveratrol(Res)is a promising anticancer drug against hepatocellular carcinoma(HCC),but whether its anti-HCC effects implicate mitophagy remains unclear.Therefore,we aimed to explore the specific role of Re...Objective Resveratrol(Res)is a promising anticancer drug against hepatocellular carcinoma(HCC),but whether its anti-HCC effects implicate mitophagy remains unclear.Therefore,we aimed to explore the specific role of Res in mitophagy and the related mechanisms during the treatment of HCC.Methods HepG2 cells and tumor-grafted nude mice were used to investigate the effects of low-,middle-and high-dose of Res on HCC progression and mitophagy in vitro and in vivo,respectively.A series of approaches including cell counting kit-8,flow cytometry,wound healing and transwell assays were used to evaluate tumor cell functions.Transmission electron microscopy,immunofluorescence and Western blotting analysis were used to assess mitophagy.Mitochondrial oxygen consumption rate,reactive oxygen species and membrane potential were used to reflect mitochondrial function.After disrupting the expression of metastasis-associated lung adenocarcinoma transcript 1(MALAT1),miR-143-3p,and ribonucleoside reductase M2(RRM2),the effects of the MALAT1/miR-143-3p/RRM2 axis on cell function and mitophagy under Res treatment were explored in vitro.Additionally,dual-luciferase reporter and chromatin immunoprecipitation were used to confirm interactions between target genes.Results Res significantly inhibited the proliferation and promoted apoptosis of HCC cells in vitro,while significantly suppressing tumor growth in a dose-dependent manner and inducing mitophagy and mitochondrial dysfunction in vivo.Interestingly,MALAT1 was highly expressed in HCC cells and its knockdown upregulated miR-143-3p expression in HCC cells,which subsequently inhibited RRM2 expression.Furthermore,in nude mice grafted with HCC tumors and treated with Res,the expression of MALAT1,miR-143-3p and RRM2 were altered significantly.In vitro data further supported the targeted binding relationships between MALAT1 and miR-143-3p and between miR-143-3p and RRM2.Therefore,a series of cell-based experiments were carried out to study the mechanism of the MALAT1/miR-143-3p/RRM2 axis involved in mitophagy and HCC;these experiments revealed that MALAT1 knockdown,miR-143-3p mimic and RRM silencing potentiated the antitumor effects of Res and its activation of mitophagy.Conclusion Res facilitated mitophagy in HCC and exerted anti-cancer effects by targeting the MALAT1/miR-143-3p/RRM2 axis.展开更多
Malate dehydrogenase(MDH)is a widely expressed enzyme that plays a key role in plant growth,development,and stress responses.However,information on MDH genes in the soybean genome is limited.Seventeen members of the s...Malate dehydrogenase(MDH)is a widely expressed enzyme that plays a key role in plant growth,development,and stress responses.However,information on MDH genes in the soybean genome is limited.Seventeen members of the soybean MDH family were identified by genome-wide analysis,and the genes were analyzed for the presence of conserved protein motifs.The genes were divided into five clusters according to their phylogenetic relationships.The intracellular localizations of six GmMDHs were determined by confocal microscopy of Arabidopsis mesophyll protoplasts.Transcripts of GmMDHs were significantly increased by abiotic stress(drought,salt,and alkalinity)and hormone treatments,as shown by an analysis of cis-regulatory elements and quantitative real-time polymerase chain reaction(qRT-PCR).The GmMDHs displayed unique expression patterns in various soybean tissues.Notably,the expression levels of a chloroplast isoform(GmMDH2)were unusually high under salt stress,presumably indicating a critical role in soybean responses to salinity.Expression of GmMDH2 in Escherichia coli showed that the recombinant enzyme has nicotinamide adenine dinucleotide phosphate(NADP)-dependent MDH activity.The redox states of the NADP(reduced form)(NADPH)pool and antioxidant activities were shown to be modulated by GmMDH2 gene overexpression,which in turn reduced reactive oxygen species(ROS)formation in transgenic soybean,significantly enhancing the salt stress resistance.Gene-based association analysis showed that variations in GmMDH2 were strongly linked to seedling salt tolerance.A polymorphism potentially associated with salt tolerance was discovered in the promoter region of GmMDH2.These findings not only improve our understanding of the stress response mechanism by identifying and characterizing the MDH gene family throughout the soybean genome but they also identified a potential candidate gene for the future enhancement of salt tolerance in soybean.展开更多
基金supported by the Zhejiang Provincial Science and Technology Department Key Research and Development Plan(No.2020C03046).
文摘Objective Resveratrol(Res)is a promising anticancer drug against hepatocellular carcinoma(HCC),but whether its anti-HCC effects implicate mitophagy remains unclear.Therefore,we aimed to explore the specific role of Res in mitophagy and the related mechanisms during the treatment of HCC.Methods HepG2 cells and tumor-grafted nude mice were used to investigate the effects of low-,middle-and high-dose of Res on HCC progression and mitophagy in vitro and in vivo,respectively.A series of approaches including cell counting kit-8,flow cytometry,wound healing and transwell assays were used to evaluate tumor cell functions.Transmission electron microscopy,immunofluorescence and Western blotting analysis were used to assess mitophagy.Mitochondrial oxygen consumption rate,reactive oxygen species and membrane potential were used to reflect mitochondrial function.After disrupting the expression of metastasis-associated lung adenocarcinoma transcript 1(MALAT1),miR-143-3p,and ribonucleoside reductase M2(RRM2),the effects of the MALAT1/miR-143-3p/RRM2 axis on cell function and mitophagy under Res treatment were explored in vitro.Additionally,dual-luciferase reporter and chromatin immunoprecipitation were used to confirm interactions between target genes.Results Res significantly inhibited the proliferation and promoted apoptosis of HCC cells in vitro,while significantly suppressing tumor growth in a dose-dependent manner and inducing mitophagy and mitochondrial dysfunction in vivo.Interestingly,MALAT1 was highly expressed in HCC cells and its knockdown upregulated miR-143-3p expression in HCC cells,which subsequently inhibited RRM2 expression.Furthermore,in nude mice grafted with HCC tumors and treated with Res,the expression of MALAT1,miR-143-3p and RRM2 were altered significantly.In vitro data further supported the targeted binding relationships between MALAT1 and miR-143-3p and between miR-143-3p and RRM2.Therefore,a series of cell-based experiments were carried out to study the mechanism of the MALAT1/miR-143-3p/RRM2 axis involved in mitophagy and HCC;these experiments revealed that MALAT1 knockdown,miR-143-3p mimic and RRM silencing potentiated the antitumor effects of Res and its activation of mitophagy.Conclusion Res facilitated mitophagy in HCC and exerted anti-cancer effects by targeting the MALAT1/miR-143-3p/RRM2 axis.
基金supported by the Natural Science Foundation of Heilongjiang Province,China(TD2022C003 and YQ2022C010)the National Key R&D Program of China(2021YFD1201104-02-02 and2021YFF1001202)the National Natural Science Foundation of China(U20A2027,31971899,32272093,and 32272072)。
文摘Malate dehydrogenase(MDH)is a widely expressed enzyme that plays a key role in plant growth,development,and stress responses.However,information on MDH genes in the soybean genome is limited.Seventeen members of the soybean MDH family were identified by genome-wide analysis,and the genes were analyzed for the presence of conserved protein motifs.The genes were divided into five clusters according to their phylogenetic relationships.The intracellular localizations of six GmMDHs were determined by confocal microscopy of Arabidopsis mesophyll protoplasts.Transcripts of GmMDHs were significantly increased by abiotic stress(drought,salt,and alkalinity)and hormone treatments,as shown by an analysis of cis-regulatory elements and quantitative real-time polymerase chain reaction(qRT-PCR).The GmMDHs displayed unique expression patterns in various soybean tissues.Notably,the expression levels of a chloroplast isoform(GmMDH2)were unusually high under salt stress,presumably indicating a critical role in soybean responses to salinity.Expression of GmMDH2 in Escherichia coli showed that the recombinant enzyme has nicotinamide adenine dinucleotide phosphate(NADP)-dependent MDH activity.The redox states of the NADP(reduced form)(NADPH)pool and antioxidant activities were shown to be modulated by GmMDH2 gene overexpression,which in turn reduced reactive oxygen species(ROS)formation in transgenic soybean,significantly enhancing the salt stress resistance.Gene-based association analysis showed that variations in GmMDH2 were strongly linked to seedling salt tolerance.A polymorphism potentially associated with salt tolerance was discovered in the promoter region of GmMDH2.These findings not only improve our understanding of the stress response mechanism by identifying and characterizing the MDH gene family throughout the soybean genome but they also identified a potential candidate gene for the future enhancement of salt tolerance in soybean.
