目的研究胰岛素样生长因子2 mRNA结合蛋白1(IGF2BP1)及长链非编码RNA LINC00160(LINC00160)在胃癌中的作用,及其可能调控胃癌细胞增殖、侵袭的潜在作用机制。方法通过实时定量聚合酶链反应(qRT-PCR)检测胃癌组织和细胞中LINC00160表达...目的研究胰岛素样生长因子2 mRNA结合蛋白1(IGF2BP1)及长链非编码RNA LINC00160(LINC00160)在胃癌中的作用,及其可能调控胃癌细胞增殖、侵袭的潜在作用机制。方法通过实时定量聚合酶链反应(qRT-PCR)检测胃癌组织和细胞中LINC00160表达水平。通过生物信息学预测及RNA结合蛋白免疫沉淀(RIP)和甲基化RNA免疫沉淀(MeRIP)法验证分析LINC00160与IGF2BP1的结合作用;Pearson检验分析胃癌组织中LINC00160与IGF2BP1表达的相关性;CCK-8法和Transwell检测细胞增殖和侵袭能力;检测分析有氧糖酵解指标[葡萄糖摄取量、乳酸生成量和三磷酸腺苷(ATP)及细胞外酸化率(ECAR)和耗氧率(OCR)]水平变化。结果与癌旁正常组织相比,胃癌组织中LINC00160(5.13±0.62 vs 1.02±0.03)表达量显著上调,差异具有统计学意义(t=-36.266,P<0.001);胃癌细胞中LINC00160表达量亦高于人正常胃上皮细胞系GES-1,差异具有统计学意义(F=24.595,P<0.001)。与对照组相比,沉默LINC00160显著抑制AGS细胞的增殖(0.42±0.03 vs 1.03±0.04)和侵袭(22.13%±1.97%vs 42.15%±2.67%),减少葡萄糖摄取(2.11±0.26mmol/L vs 4.22±0.37mmol/L)和乳酸生成量(6.84±1.25mmol/L vs 11.68±1.55mmol/L),降低ECAR,升高ATP(3.34±0.29mmol/L vs 1.87±0.24mmol/L)水平和OCR,差异具有统计学意义(t=4.188~24.423,均P<0.01)。胃癌组织中IGF2BP1蛋白(4.07±0.36)表达显著高于癌旁组织(1.01±0.03),差异具有统计学意义(t=-46.396,P<0.01),且与LINC00160表达呈正相关(r 2=0.7745,P<0.01)。IGF2BP1通过与LINC00160的m6A修饰位点结合促进LINC00160稳定性。沉默IGF2BP1显著抑制了LINC00160表达及胃癌细胞的增殖、侵袭和有氧糖酵解,差异具有统计学意义(t=4.386~11.989,均P<0.01);过表达LINC00160可逆转沉默IGF2BP1对胃癌细胞生物学行为及有氧糖酵解的影响。结论胃癌中LINC00160显著上调,IGF2BP1可能通过m6A修饰稳定调控LINC00160表达,促进肿瘤细胞的有氧糖酵解,参与胃癌的发生发展。展开更多
Multiple myeloma(MM)patients with chromosome 1q gain(1q+)are clinically and biologically heterogeneous.The underlying molecular mechanisms are still under investigation,while the identification of targets for effectiv...Multiple myeloma(MM)patients with chromosome 1q gain(1q+)are clinically and biologically heterogeneous.The underlying molecular mechanisms are still under investigation,while the identification of targets for effective therapy of this subgroup of MM patients is urgently needed.We aimed to investigate the clinical significance and the regulatory mechanisms of insulin-like growth factor 2 messenger RNA(mRNA)binding protein 1(IGF2BP1),a N6-methyladenosine(m6A)reader,in MM patients with 1q+.We found that MM patients with 1q+exhibit a significantly higher level of IGF2BP1 mRNA than controls,while higher IGF2BP1 expression predicted a worse prognosis in MM patients with 1q+.IGF2BP1 overexpression promoted cell proliferation and G1-to-S phase transition of the cell cycle in NCI-H929 cells.Through comprehensive in silico analyses of existing public datasets and in-house generated high-throughput sequencing datasets,along with in vitro experiments,we identified CDC5L as a target of IGFBP1,which can bind to the m6A sites of CDC5L mRNA to up-regulate its protein abundance.Higher CDC5L expression also predicted a worse prognosis of MM patients with 1q+.Moreover,both knockdown and mutation of CDC5L attenuated the pro-proliferative effect of IGF2BP1.Furthermore,IGF2BP1 inhibitor BTYNB effectively inhibited CDC5L expression in MM cells with 1q+and suppressed the proliferation of these cells in vitro and in vivo.Therefore,IGF2BP1 acts as a post-transcriptional enhancer of CDC5L in an m6A-dependent manner to promote the proliferation of MM cells with 1q+.Our work identified a novel IGF2BP1-CDC5L axis and provided new insight into developing targeted therapeutics for MM patients with 1q+.展开更多
N6-methyladenosine(m6A)is the most abundant and well-investigated internal RNA modification in eukaryotic RNAs,affecting its target gene expression by controlling RNA localization,splicing,stability,and translation.m6...N6-methyladenosine(m6A)is the most abundant and well-investigated internal RNA modification in eukaryotic RNAs,affecting its target gene expression by controlling RNA localization,splicing,stability,and translation.m6A modifications are regulated by m6A methyltransferase complex,demethylase,and reading proteins.Insulin-like growth factor-2 mRNA-binding protein 1(IGF2BP1),a member of a conserved family of single-stranded RNA-binding proteins,has recently been identified as a vital m6A reading protein.IGF2BP1 is highly expressed in various tumors and is associated with poor prognosis and treatment resistance.Furthermore,previous studies have shown that IGF2BP1 plays critical roles in regulating various cancer hallmarks,including sustained cell proliferation,cell death resistance,activation of invasion and metastasis,deregulated cellular energetics,immune evasion,and unlocking phenotypic plasticity.IGF2BP1 could promote the expression of cancer-related genes by recognizing their m6A sites,thereby altering cell characteristics,and eventually,malignancy.Therefore,IGF2BP1 might be a potential target for tumor diagnosis and anti-tumor therapeutic strategies.This review summarizes the current knowledge on the functional roles and underlying molecular mechanisms of IGF2BP1 in regulating cancer hallmarks.Moreover,we discuss the prospects of IGF2BP1 as a potential tumor diagnosis marker,as well as a potential target for an anti-tumor therapeutic strategy.展开更多
文摘目的研究胰岛素样生长因子2 mRNA结合蛋白1(IGF2BP1)及长链非编码RNA LINC00160(LINC00160)在胃癌中的作用,及其可能调控胃癌细胞增殖、侵袭的潜在作用机制。方法通过实时定量聚合酶链反应(qRT-PCR)检测胃癌组织和细胞中LINC00160表达水平。通过生物信息学预测及RNA结合蛋白免疫沉淀(RIP)和甲基化RNA免疫沉淀(MeRIP)法验证分析LINC00160与IGF2BP1的结合作用;Pearson检验分析胃癌组织中LINC00160与IGF2BP1表达的相关性;CCK-8法和Transwell检测细胞增殖和侵袭能力;检测分析有氧糖酵解指标[葡萄糖摄取量、乳酸生成量和三磷酸腺苷(ATP)及细胞外酸化率(ECAR)和耗氧率(OCR)]水平变化。结果与癌旁正常组织相比,胃癌组织中LINC00160(5.13±0.62 vs 1.02±0.03)表达量显著上调,差异具有统计学意义(t=-36.266,P<0.001);胃癌细胞中LINC00160表达量亦高于人正常胃上皮细胞系GES-1,差异具有统计学意义(F=24.595,P<0.001)。与对照组相比,沉默LINC00160显著抑制AGS细胞的增殖(0.42±0.03 vs 1.03±0.04)和侵袭(22.13%±1.97%vs 42.15%±2.67%),减少葡萄糖摄取(2.11±0.26mmol/L vs 4.22±0.37mmol/L)和乳酸生成量(6.84±1.25mmol/L vs 11.68±1.55mmol/L),降低ECAR,升高ATP(3.34±0.29mmol/L vs 1.87±0.24mmol/L)水平和OCR,差异具有统计学意义(t=4.188~24.423,均P<0.01)。胃癌组织中IGF2BP1蛋白(4.07±0.36)表达显著高于癌旁组织(1.01±0.03),差异具有统计学意义(t=-46.396,P<0.01),且与LINC00160表达呈正相关(r 2=0.7745,P<0.01)。IGF2BP1通过与LINC00160的m6A修饰位点结合促进LINC00160稳定性。沉默IGF2BP1显著抑制了LINC00160表达及胃癌细胞的增殖、侵袭和有氧糖酵解,差异具有统计学意义(t=4.386~11.989,均P<0.01);过表达LINC00160可逆转沉默IGF2BP1对胃癌细胞生物学行为及有氧糖酵解的影响。结论胃癌中LINC00160显著上调,IGF2BP1可能通过m6A修饰稳定调控LINC00160表达,促进肿瘤细胞的有氧糖酵解,参与胃癌的发生发展。
基金supported by the National Natural Science Foundation of China(No.82100215)Natural Science Foundation of Shanghai,China(No.22ZR1411400)Natural Science Foundation of Fujian Province,China(No.2023J05291).
文摘Multiple myeloma(MM)patients with chromosome 1q gain(1q+)are clinically and biologically heterogeneous.The underlying molecular mechanisms are still under investigation,while the identification of targets for effective therapy of this subgroup of MM patients is urgently needed.We aimed to investigate the clinical significance and the regulatory mechanisms of insulin-like growth factor 2 messenger RNA(mRNA)binding protein 1(IGF2BP1),a N6-methyladenosine(m6A)reader,in MM patients with 1q+.We found that MM patients with 1q+exhibit a significantly higher level of IGF2BP1 mRNA than controls,while higher IGF2BP1 expression predicted a worse prognosis in MM patients with 1q+.IGF2BP1 overexpression promoted cell proliferation and G1-to-S phase transition of the cell cycle in NCI-H929 cells.Through comprehensive in silico analyses of existing public datasets and in-house generated high-throughput sequencing datasets,along with in vitro experiments,we identified CDC5L as a target of IGFBP1,which can bind to the m6A sites of CDC5L mRNA to up-regulate its protein abundance.Higher CDC5L expression also predicted a worse prognosis of MM patients with 1q+.Moreover,both knockdown and mutation of CDC5L attenuated the pro-proliferative effect of IGF2BP1.Furthermore,IGF2BP1 inhibitor BTYNB effectively inhibited CDC5L expression in MM cells with 1q+and suppressed the proliferation of these cells in vitro and in vivo.Therefore,IGF2BP1 acts as a post-transcriptional enhancer of CDC5L in an m6A-dependent manner to promote the proliferation of MM cells with 1q+.Our work identified a novel IGF2BP1-CDC5L axis and provided new insight into developing targeted therapeutics for MM patients with 1q+.
基金supported by grants from the National Natural Science Foundation of China(No.82173029,32270778,and 82372655)the Natural Science Foundation of Chongqing,China(No.CSTB2022NSCQ-MSX0611,CSTB2022NSCQ-MSX0612)the Talent Project of Chongqing University Jiangjin Hospital(Chongqing,China)(No.2024LJXM005).
文摘N6-methyladenosine(m6A)is the most abundant and well-investigated internal RNA modification in eukaryotic RNAs,affecting its target gene expression by controlling RNA localization,splicing,stability,and translation.m6A modifications are regulated by m6A methyltransferase complex,demethylase,and reading proteins.Insulin-like growth factor-2 mRNA-binding protein 1(IGF2BP1),a member of a conserved family of single-stranded RNA-binding proteins,has recently been identified as a vital m6A reading protein.IGF2BP1 is highly expressed in various tumors and is associated with poor prognosis and treatment resistance.Furthermore,previous studies have shown that IGF2BP1 plays critical roles in regulating various cancer hallmarks,including sustained cell proliferation,cell death resistance,activation of invasion and metastasis,deregulated cellular energetics,immune evasion,and unlocking phenotypic plasticity.IGF2BP1 could promote the expression of cancer-related genes by recognizing their m6A sites,thereby altering cell characteristics,and eventually,malignancy.Therefore,IGF2BP1 might be a potential target for tumor diagnosis and anti-tumor therapeutic strategies.This review summarizes the current knowledge on the functional roles and underlying molecular mechanisms of IGF2BP1 in regulating cancer hallmarks.Moreover,we discuss the prospects of IGF2BP1 as a potential tumor diagnosis marker,as well as a potential target for an anti-tumor therapeutic strategy.
