[目的]探究FBXO22调控ZEB1对肝细胞癌增殖、侵袭的影响。[方法]收集39例HCC患者的癌组织和癌旁组织。将Hep G2细胞随机分为4组:阴性对照组(加入0.5μmol/L si NC处理),si FBXO22组(加入0.5μmol/L si FBXO22处理),si ZEB1组(加入0.5μmo...[目的]探究FBXO22调控ZEB1对肝细胞癌增殖、侵袭的影响。[方法]收集39例HCC患者的癌组织和癌旁组织。将Hep G2细胞随机分为4组:阴性对照组(加入0.5μmol/L si NC处理),si FBXO22组(加入0.5μmol/L si FBXO22处理),si ZEB1组(加入0.5μmol/L si ZEB1处理),si FBXO22+si ZEB1组(加入0.5μmol/L si FBXO22及si ZEB1处理)。通过免疫组化分析FBXO22的表达水平,MTT、Transwell、流式细胞术、Western blot分别检测细胞增殖、侵袭、凋亡、蛋白表达水平。[结果]与癌旁组织相比,HCC组织中FBXO22表达增加(P<0.05);与阴性对照组比较,si FBXO22组、si ZEB1组、si FBXO22+si ZEB1组的Hep G2细胞增殖、侵袭能力降低(P<0.05)、凋亡率增加(P<0.05)、ZEB1蛋白表达降低(P<0.05)。[结论]FBXO22在肝细胞癌组织中高表达,下调FBXO22的表达后能够抑制ZEB1进而减弱肝细胞癌Hep G2细胞的增殖、侵袭活性,并促进Hep G2细胞凋亡。展开更多
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
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.
