BACKGROUND N6-methyladenosine(m6A)exerts a pro-carcinogenic effect in diverse cancers.The relationship between m6A-reading protein IGF2BP3 and gastric cancer(GC)has not yet been fully elucidated.AIM To investigate the...BACKGROUND N6-methyladenosine(m6A)exerts a pro-carcinogenic effect in diverse cancers.The relationship between m6A-reading protein IGF2BP3 and gastric cancer(GC)has not yet been fully elucidated.AIM To investigate the molecular mechanisms of IGF2BP3 in GC carcinogenesis and progression and thus provide a rationale for novel therapeutic strategies.METHODS Expression levels of IGF2BP3 in GC were determined using quantitative reverse transcription polymerase chain reaction(qRT-PCR),western blot(WB),and immunohistochemistry,and their associations with patients’clinicopathological characteristics were analyzed.The role of IGF2BP3 in GC was investigated using cellular functional assays and subcutaneous xenograft models,and its downstream targets and signaling pathways were identified using highthroughput sequencing,bioinformatics analysis,RNA immunoprecipitation qPCR,dual luciferase reporter assay,qRT-PCR,and WB.The mechanism of IGF2BP3 in GC was validated via WB and rescue and inhibition experiments.RESULTS IGF2BP3 was highly expressed in GC and associated with diffuse-type GC,incidence of lymph node metastasis,advanced tumor node metastasis stage,and deeper tumor invasion depth.In vitro experiments demonstrated that IGF2BP3 promoted proliferation,migration,and invasiveness of GC cells,while inhibiting apoptosis and augmenting intracellular levels of glucose metabolism.In vivo experiments revealed that IGF2BP3 contributes to the growth of GC.Mechanistically,IGF2BP3 recognized and bound to the m6A site at position 1427 on FBXO32 messenger RNA,thereby increasing protein expression of FBXO32,and further activated the downstream cyclic guanosine monophosphate-protein kinase G(cGMP-PKG)signaling pathway to modulate various biological functions of GC cells and promote progression of GC.Furthermore,treatment with a selective PKG inhibitor KT5823 significantly suppressed the proliferative capacity of GC cells.CONCLUSION IGF2BP3 increases FBXO32 protein expression in an m6A-dependent manner,activates the cGMP-PKG signaling pathway,and promotes GC progression.Targeting of the IGF2BP3/FBXO32/cGMP-PKG axis could thus represent a promising therapeutic modality for GC.展开更多
为了分析甘南牦牛(Bos grunniens)肌肉萎缩盒蛋白32(F-box protein 32,FBXO32)基因的单核苷酸多态性(single nucleotide polymorphism,SNP)位点,以及基因型与胴体和肉质性状间的相关性,本研究以593头甘南牦牛为研究对象,采用混池测序和...为了分析甘南牦牛(Bos grunniens)肌肉萎缩盒蛋白32(F-box protein 32,FBXO32)基因的单核苷酸多态性(single nucleotide polymorphism,SNP)位点,以及基因型与胴体和肉质性状间的相关性,本研究以593头甘南牦牛为研究对象,采用混池测序和竞争性等位基因特异性PCR(kompetitive allele specific PCR,KASP)技术,检测了甘南牦牛FBXO32基因突变位点及基因型,分析了基因型与甘南牦牛胴体及肉质性状的相关性。结果表明,从甘南牦牛FBXO32基因检测到7个SNP位点,分别是位于5′UTR区的SNP1(g.267A>C)、外显子1区的SNP2(g.326G>T)、外显子8区的SNP3(g.31231G>C),以及3′UTR区的SNP4(g.31352G>A)、SNP5(g.31424C>T)、SNP6(g.31503A>C)和SNP7(g.31504A>G)。其中,SNP1、SNP4、SNP5、SNP6与肌肉嫩度显著相关(P<0.05),SNP3、SNP6、SNP7与失水率显著相关(P<0.05),SNP1、SNP2、SNP3、SNP4、SNP5、SNP7与眼肌面积显著相关(P<0.05),SNP2、SNP5、SNP6与胴体重显著相关(P<0.05)。7个SNP位点构建的9种有效单倍型组合中,单倍型组合H2H2的个体肌肉嫩度小、熟肉率高,单倍型组合H1H2的个体熟肉率高、眼肌面积大、失水率低。FBXO32基因突变位点可作为甘南牦牛胴体及肉质性状潜在分子遗传标记,研究结果丰富了甘南牦牛经济性状分子遗传理论基础。展开更多
基金Supported by the Hebei Natural Science Foundation,No.H2025206524,No.H2022206292 and No.H2024206140Hebei Provincial Government-funded Provincial Medical Excellent Talent Project,No.ZF2023025,No.ZF2024134 and No.LS202008+3 种基金Key RD Program of Hebei Province,No.223777103D and No.223777113DPrevention and Treatment of Geriatric Diseases by Hebei Provincial Department of Finance,No.LNB202202 and No.LNB201909Spark Scientific Research Project of the First Hospital of Hebei Medical University,No.XH202504,No.XH202312 and No.XH201805Hebei Province Medical Applicable Technology Tracking Project,No.G2019035.
文摘BACKGROUND N6-methyladenosine(m6A)exerts a pro-carcinogenic effect in diverse cancers.The relationship between m6A-reading protein IGF2BP3 and gastric cancer(GC)has not yet been fully elucidated.AIM To investigate the molecular mechanisms of IGF2BP3 in GC carcinogenesis and progression and thus provide a rationale for novel therapeutic strategies.METHODS Expression levels of IGF2BP3 in GC were determined using quantitative reverse transcription polymerase chain reaction(qRT-PCR),western blot(WB),and immunohistochemistry,and their associations with patients’clinicopathological characteristics were analyzed.The role of IGF2BP3 in GC was investigated using cellular functional assays and subcutaneous xenograft models,and its downstream targets and signaling pathways were identified using highthroughput sequencing,bioinformatics analysis,RNA immunoprecipitation qPCR,dual luciferase reporter assay,qRT-PCR,and WB.The mechanism of IGF2BP3 in GC was validated via WB and rescue and inhibition experiments.RESULTS IGF2BP3 was highly expressed in GC and associated with diffuse-type GC,incidence of lymph node metastasis,advanced tumor node metastasis stage,and deeper tumor invasion depth.In vitro experiments demonstrated that IGF2BP3 promoted proliferation,migration,and invasiveness of GC cells,while inhibiting apoptosis and augmenting intracellular levels of glucose metabolism.In vivo experiments revealed that IGF2BP3 contributes to the growth of GC.Mechanistically,IGF2BP3 recognized and bound to the m6A site at position 1427 on FBXO32 messenger RNA,thereby increasing protein expression of FBXO32,and further activated the downstream cyclic guanosine monophosphate-protein kinase G(cGMP-PKG)signaling pathway to modulate various biological functions of GC cells and promote progression of GC.Furthermore,treatment with a selective PKG inhibitor KT5823 significantly suppressed the proliferative capacity of GC cells.CONCLUSION IGF2BP3 increases FBXO32 protein expression in an m6A-dependent manner,activates the cGMP-PKG signaling pathway,and promotes GC progression.Targeting of the IGF2BP3/FBXO32/cGMP-PKG axis could thus represent a promising therapeutic modality for GC.
文摘为了分析甘南牦牛(Bos grunniens)肌肉萎缩盒蛋白32(F-box protein 32,FBXO32)基因的单核苷酸多态性(single nucleotide polymorphism,SNP)位点,以及基因型与胴体和肉质性状间的相关性,本研究以593头甘南牦牛为研究对象,采用混池测序和竞争性等位基因特异性PCR(kompetitive allele specific PCR,KASP)技术,检测了甘南牦牛FBXO32基因突变位点及基因型,分析了基因型与甘南牦牛胴体及肉质性状的相关性。结果表明,从甘南牦牛FBXO32基因检测到7个SNP位点,分别是位于5′UTR区的SNP1(g.267A>C)、外显子1区的SNP2(g.326G>T)、外显子8区的SNP3(g.31231G>C),以及3′UTR区的SNP4(g.31352G>A)、SNP5(g.31424C>T)、SNP6(g.31503A>C)和SNP7(g.31504A>G)。其中,SNP1、SNP4、SNP5、SNP6与肌肉嫩度显著相关(P<0.05),SNP3、SNP6、SNP7与失水率显著相关(P<0.05),SNP1、SNP2、SNP3、SNP4、SNP5、SNP7与眼肌面积显著相关(P<0.05),SNP2、SNP5、SNP6与胴体重显著相关(P<0.05)。7个SNP位点构建的9种有效单倍型组合中,单倍型组合H2H2的个体肌肉嫩度小、熟肉率高,单倍型组合H1H2的个体熟肉率高、眼肌面积大、失水率低。FBXO32基因突变位点可作为甘南牦牛胴体及肉质性状潜在分子遗传标记,研究结果丰富了甘南牦牛经济性状分子遗传理论基础。
