A complex network of factors contributes to neuroinflammation,such as infections,brain injuries and accumulation of toxic metabolites（Gendelman,2002）.Eicosanoids and several cytokines are the main mediators of infla...A complex network of factors contributes to neuroinflammation,such as infections,brain injuries and accumulation of toxic metabolites（Gendelman,2002）.Eicosanoids and several cytokines are the main mediators of inflammatory process;in fact,when an inflammatory condition persists,it can be responsible for the progression of degenerative diseases,展开更多
BACKGROUND Increasing data indicated that long noncoding RNAs(lncRNAs)were directly or indirectly involved in the occurrence and development of tumors,including hepatocellular carcinoma(HCC).Recent studies had found t...BACKGROUND Increasing data indicated that long noncoding RNAs(lncRNAs)were directly or indirectly involved in the occurrence and development of tumors,including hepatocellular carcinoma(HCC).Recent studies had found that the expression of lncRNA HAND2-AS1 was downregulated in HCC tissues,but its role in HCC progression is unclear.Ultrasound targeted microbubble destruction mediated gene transfection is a new method to overexpress genes.AIM To study the role of ultrasound microbubbles(UTMBs)mediated HAND2-AS1 in the progression of HCC,in order to provide a new reference for the treatment of HCC.METHODS In vitro,we transfected HAND2-AS1 siRNA into HepG2 cells by UTMBs,and detected cell proliferation,apoptosis,invasion and epithelial-mesenchymal transition(EMT)by cell counting kit-8 assay,flow cytometry,Transwell invasion assay and Western blotting,respectively.In addition,we transfected miR-837-5p mimic into UTMBs treated cells and observed the changes of cell behavior.Next,the UTMBs treated HepG2 cells were transfected together with miR-837-5p mimic and tissue inhibitor of matrix metalloproteinase-2(TIMP2)overexpression vector,and we detected cell proliferation,apoptosis,invasion and EMT.In vivo,we established a mouse model of subcutaneous transplantation of HepG2 cells and observed the effect of HAND2-AS1 silencing on tumor formation ability.RESULTS We found that UTMBs carrying HAND2-AS1 restricted cell proliferation,invasion,and EMT,encouraged apoptosis,and HAND2-AS1 silencing eliminated the effect of UTMBs.Additionally,miR-873-5p targets the gene HAND2-AS1,which also targets the 3’UTR of TIMP2.And miR-873-5p mimic counteracted the impact of HAND2-AS1.Further,miR-873-5p mimic solely or in combination with pcDNA-TIMP2 had been transformed into HepG2 cells exposed to UTMBs.We discovered that TIMP2 reversed the effect of miR-873-5p mimic caused by the blocked signalling cascade for matrix metalloproteinase(MMP)2/MMP9.In vivo results showed that HAND2-AS1 silencing significantly inhibited tumor formation in mice.CONCLUSION LncRNA HAND2-AS1 promotes TIMP2 expression by targeting miR-873-5p to inhibit HepG2 cell growth and delay HCC progression.展开更多
Divisions at the periphery and midzone of mitochondria are two fission signatures that determine the fate of mitochondria and cells.Pharmacological induction of excessively asymmetric mitofissionassociated cell death(...Divisions at the periphery and midzone of mitochondria are two fission signatures that determine the fate of mitochondria and cells.Pharmacological induction of excessively asymmetric mitofissionassociated cell death(MFAD)by switching the scission position from the mitochondrial midzone to the periphery represents a promising strategy for anticancer therapy.By screening a series of paninhibitors,we identified pracinostat,a pan-histone deacetylase(HDAC)inhibitor,as a novel MFAD inducer,that exhibited a significant anticancer effect on colorectal cancer(CRC)in vivo and in vitro.Pracinostat increased the expression of cyclin-dependent kinase 5(CDK5)and induced its acetylation at residue lysine 33,accelerating the formation of complex CDK5/CDK5 regulatory subunit 1 and dynaminrelated protein 1(Drp1)-mediated mitochondrial peripheral fission.CRC cells with high level of CDK5(CDK5-high)displayed midzone mitochondrial division that was associated with oncogenic phenotype,but treatment with pracinostat led to a lethal increase in the already-elevated level of CDK5 in the CRC cells.Mechanistically,pracinostat switched the scission position from the mitochondrial midzone to the periphery by improving the binding of Drp1 from mitochondrial fission factor(MFF)to mitochondrial fission 1 protein(FIS1).Thus,our results revealed the anticancer mechanism of HDACi pracinostat in CRC via activating CDK5-Drp1 signaling to cause selective MFAD of those CDK5-high tumor cells,which implicates a new paradigm to develop potential therapeutic strategies for CRC treatment.展开更多
生长抑制蛋白5(inhibitor of growth protein 5,ING5)是生长抑制蛋白家族的成员之一,参与调节细胞周期、细胞增殖和凋亡等多种生命活动。家蚕核型多角体病毒(Bombyx mori nucleopolyhedrovirus,BmNPV)侵染家蚕卵巢细胞BmN前后的蛋白质...生长抑制蛋白5(inhibitor of growth protein 5,ING5)是生长抑制蛋白家族的成员之一,参与调节细胞周期、细胞增殖和凋亡等多种生命活动。家蚕核型多角体病毒(Bombyx mori nucleopolyhedrovirus,BmNPV)侵染家蚕卵巢细胞BmN前后的蛋白质乙酰化修饰差异组学分析结果显示,BmN细胞中的ING5蛋白有3个赖氨酸残基位点(K136、K137和K154)的乙酰化修饰水平在BmNPV感染后显著下调。为了探究ING5乙酰化修饰对其功能的影响以及在BmNPV侵染过程中的调控作用机制,首先克隆了家蚕ING5基因,并将BmNPV感染后乙酰化修饰水平显著下调的赖氨酸(K)定点突变为谷氨酰胺(Q)以模拟乙酰化修饰,突变为精氨酸(R)以模拟去乙酰化修饰。然后构建瞬时表达载体并转染BmN细胞,结果显示ING5蛋白过表达具有显著抑制细胞活力的作用,而ING5乙酰化修饰则可以显著提高细胞活力。进一步研究发现,过表达ING5蛋白具有显著促细胞凋亡作用,而ING5乙酰化修饰则显著抑制细胞凋亡。酵母双杂交试验结果显示,野生型ING5与凋亡相关蛋白P53可以互作,但ING5的乙酰化修饰影响了该互作关系;同时还发现ING5的乙酰化可显著降低P53蛋白稳定性。上述结果表明,家蚕ING5可能通过P53依赖的方式参与细胞凋亡调控,但K136、K137和K154位点的乙酰化修饰改变了ING5与P53的相互作用,进而影响细胞凋亡。研究结果将为深入解析家蚕ING5家族蛋白调控BmNPV侵染的作用机制提供试验依据,同时也可为家蚕的抗病毒育种提供新的理论依据。展开更多
文摘A complex network of factors contributes to neuroinflammation,such as infections,brain injuries and accumulation of toxic metabolites（Gendelman,2002）.Eicosanoids and several cytokines are the main mediators of inflammatory process;in fact,when an inflammatory condition persists,it can be responsible for the progression of degenerative diseases,
文摘BACKGROUND Increasing data indicated that long noncoding RNAs(lncRNAs)were directly or indirectly involved in the occurrence and development of tumors,including hepatocellular carcinoma(HCC).Recent studies had found that the expression of lncRNA HAND2-AS1 was downregulated in HCC tissues,but its role in HCC progression is unclear.Ultrasound targeted microbubble destruction mediated gene transfection is a new method to overexpress genes.AIM To study the role of ultrasound microbubbles(UTMBs)mediated HAND2-AS1 in the progression of HCC,in order to provide a new reference for the treatment of HCC.METHODS In vitro,we transfected HAND2-AS1 siRNA into HepG2 cells by UTMBs,and detected cell proliferation,apoptosis,invasion and epithelial-mesenchymal transition(EMT)by cell counting kit-8 assay,flow cytometry,Transwell invasion assay and Western blotting,respectively.In addition,we transfected miR-837-5p mimic into UTMBs treated cells and observed the changes of cell behavior.Next,the UTMBs treated HepG2 cells were transfected together with miR-837-5p mimic and tissue inhibitor of matrix metalloproteinase-2(TIMP2)overexpression vector,and we detected cell proliferation,apoptosis,invasion and EMT.In vivo,we established a mouse model of subcutaneous transplantation of HepG2 cells and observed the effect of HAND2-AS1 silencing on tumor formation ability.RESULTS We found that UTMBs carrying HAND2-AS1 restricted cell proliferation,invasion,and EMT,encouraged apoptosis,and HAND2-AS1 silencing eliminated the effect of UTMBs.Additionally,miR-873-5p targets the gene HAND2-AS1,which also targets the 3’UTR of TIMP2.And miR-873-5p mimic counteracted the impact of HAND2-AS1.Further,miR-873-5p mimic solely or in combination with pcDNA-TIMP2 had been transformed into HepG2 cells exposed to UTMBs.We discovered that TIMP2 reversed the effect of miR-873-5p mimic caused by the blocked signalling cascade for matrix metalloproteinase(MMP)2/MMP9.In vivo results showed that HAND2-AS1 silencing significantly inhibited tumor formation in mice.CONCLUSION LncRNA HAND2-AS1 promotes TIMP2 expression by targeting miR-873-5p to inhibit HepG2 cell growth and delay HCC progression.
基金supported by the National Natural Science Foundation of China(Grant Nos.:82103208,and 82002948)the Guangdong Basic and Applied Basic Research Foundation(Grant Nos.:2022A1515220212,and 2023A1515030115)+1 种基金National Key R&D Program of China(Grant No.:2020YFE0202200)Jinan University National College Students'Innovation and Entrepreneurship Training Program(Program No.:202110559085).
文摘Divisions at the periphery and midzone of mitochondria are two fission signatures that determine the fate of mitochondria and cells.Pharmacological induction of excessively asymmetric mitofissionassociated cell death(MFAD)by switching the scission position from the mitochondrial midzone to the periphery represents a promising strategy for anticancer therapy.By screening a series of paninhibitors,we identified pracinostat,a pan-histone deacetylase(HDAC)inhibitor,as a novel MFAD inducer,that exhibited a significant anticancer effect on colorectal cancer(CRC)in vivo and in vitro.Pracinostat increased the expression of cyclin-dependent kinase 5(CDK5)and induced its acetylation at residue lysine 33,accelerating the formation of complex CDK5/CDK5 regulatory subunit 1 and dynaminrelated protein 1(Drp1)-mediated mitochondrial peripheral fission.CRC cells with high level of CDK5(CDK5-high)displayed midzone mitochondrial division that was associated with oncogenic phenotype,but treatment with pracinostat led to a lethal increase in the already-elevated level of CDK5 in the CRC cells.Mechanistically,pracinostat switched the scission position from the mitochondrial midzone to the periphery by improving the binding of Drp1 from mitochondrial fission factor(MFF)to mitochondrial fission 1 protein(FIS1).Thus,our results revealed the anticancer mechanism of HDACi pracinostat in CRC via activating CDK5-Drp1 signaling to cause selective MFAD of those CDK5-high tumor cells,which implicates a new paradigm to develop potential therapeutic strategies for CRC treatment.
文摘生长抑制蛋白5(inhibitor of growth protein 5,ING5)是生长抑制蛋白家族的成员之一,参与调节细胞周期、细胞增殖和凋亡等多种生命活动。家蚕核型多角体病毒(Bombyx mori nucleopolyhedrovirus,BmNPV)侵染家蚕卵巢细胞BmN前后的蛋白质乙酰化修饰差异组学分析结果显示,BmN细胞中的ING5蛋白有3个赖氨酸残基位点(K136、K137和K154)的乙酰化修饰水平在BmNPV感染后显著下调。为了探究ING5乙酰化修饰对其功能的影响以及在BmNPV侵染过程中的调控作用机制,首先克隆了家蚕ING5基因,并将BmNPV感染后乙酰化修饰水平显著下调的赖氨酸(K)定点突变为谷氨酰胺(Q)以模拟乙酰化修饰,突变为精氨酸(R)以模拟去乙酰化修饰。然后构建瞬时表达载体并转染BmN细胞,结果显示ING5蛋白过表达具有显著抑制细胞活力的作用,而ING5乙酰化修饰则可以显著提高细胞活力。进一步研究发现,过表达ING5蛋白具有显著促细胞凋亡作用,而ING5乙酰化修饰则显著抑制细胞凋亡。酵母双杂交试验结果显示,野生型ING5与凋亡相关蛋白P53可以互作,但ING5的乙酰化修饰影响了该互作关系;同时还发现ING5的乙酰化可显著降低P53蛋白稳定性。上述结果表明,家蚕ING5可能通过P53依赖的方式参与细胞凋亡调控,但K136、K137和K154位点的乙酰化修饰改变了ING5与P53的相互作用,进而影响细胞凋亡。研究结果将为深入解析家蚕ING5家族蛋白调控BmNPV侵染的作用机制提供试验依据,同时也可为家蚕的抗病毒育种提供新的理论依据。
