目的:目前尚不清楚m6A甲基化基因在结直肠癌分化中的作用。本研究结合生物信息学分析与实验验证,探讨这些基因在结直肠癌中的表达特征及其潜在机制。方法:利用生物信息学,我们发现YTHDF1、YTHDF2、ALKBH5、WTAP与结直肠癌表达异常显著(p...目的:目前尚不清楚m6A甲基化基因在结直肠癌分化中的作用。本研究结合生物信息学分析与实验验证,探讨这些基因在结直肠癌中的表达特征及其潜在机制。方法:利用生物信息学,我们发现YTHDF1、YTHDF2、ALKBH5、WTAP与结直肠癌表达异常显著(p Background: The role of m6A methylation genes in the differentiation of colorectal cancer is currently unclear. This study combines bioinformatics analysis and experimental validation to explore the expression patterns of these genes in colorectal cancer and their potential mechanisms. Methods: Using bioinformatics, we found that YTHDF1, YTHDF2, ALKBH5, and WTAP are significantly aberrantly expressed in colorectal cancer (p < 0.05). In a real-world cohort from Lishui, quantitative PCR (qPCR) was used to measure mRNA expression of these four genes in CRC tissues and adjacent normal tissues. Multivariable logistic regression analysis with SPSS identified YTHDF1 and WTAP mRNA expression levels as being associated with CRC differentiation. Immunohistochemistry (IHC) was performed to examine corresponding protein levels, quantified using ImageJ. Results showed that YTHDF1 protein expression negatively correlated with CRC differentiation, whereas WTAP protein expression positively correlated. Additionally, multivariable Cox regression analysis was conducted to identify potential factors influencing CRC development. Results: In Lishui, YTHDF1 protein expression was negatively associated with CRC differentiation, while WTAP protein expression was positively associated. Age and alcohol consumption were key risk factors for CRC development, while no significant associations were found with sex or smoking. GO enrichment analysis suggests that YTHDF1 and WTAP may regulate the differentiation process of colorectal cancer by participating in mechanisms such as mRNA modification, translation regulation, and RNA stability. For the poorly differentiated CRC group, time to progression rate (TTP) was 100% and overall survival rate (OS) was 0% within three years. Conclusions: In Lishui City, the YTHDF1 and WTAP genes influence the differentiation of colorectal cancer and may serve as important therapeutic targets and biomarkers for the diagnosis of patients with colorectal cancer.展开更多
N6-methyladenosine(m^(6)A)modification of mRNA is a critical post-transcriptional regulatory mechanism that modulates mRNA metabolism and neuronal function.The m^(6)A reader YTHDF1 has been shown to enhance the transl...N6-methyladenosine(m^(6)A)modification of mRNA is a critical post-transcriptional regulatory mechanism that modulates mRNA metabolism and neuronal function.The m^(6)A reader YTHDF1 has been shown to enhance the translational efficiency of m^(6)A-modified mRNAs in the brain and is essential for learning and memory.However,its role in the mature retina remains unclear.Herein,we report a novel role of Ythdf1 in the maintenance of retinal function using a genetic knockout model.Loss of Ythdf1 resulted in impaired scotopic electroretinogram(ERG)responses and progressive retinal degeneration.Detailed analyses of rod photoreceptors confirmed substantial degenerative changes in the absence of ciliary defects.Single-cell RNA sequencing revealed comprehensive molecular alterations across all retinal cell types in Ythdf1-deficient retinas.Integrative analysis of methylated RNA immunoprecipitation(MeRIP)sequencing and RIP sequencing identified Tulp1 and Dhx38,two inheritable retinal degeneration disease-associated gene homologs,as direct targets of YTHDF1 in the retina.Specifically,YTHDF1 recognized and bound m^(6)A-modified Tulp1 and Dhx38 mRNA at the coding sequence(CDS),enhancing their translational efficiency without altering mRNA levels.Collectively,these findings highlight the essential role of YTHDF1 in preserving visual function and reveal a novel regulatory mechanism of m^(6)A reader proteins in retinal degeneration,identifying potential therapeutic targets for severe retinopathies.展开更多
Background Intramuscular fat(IMF)content is a critical indicator of pork quality,and abnormal IMF is also relevant to human disease as well as aging.Although N6-methyladenosine(m^(6)A)RNA modification was recently fou...Background Intramuscular fat(IMF)content is a critical indicator of pork quality,and abnormal IMF is also relevant to human disease as well as aging.Although N6-methyladenosine(m^(6)A)RNA modification was recently found to regulate adipogenesis in porcine intramuscular fat,however,the underlying molecular mechanisms was still unclear.Results In this work,we collected 20 longissimus dorsi muscle samples with high(average 3.95%)or low IMF content(average 1.22%)from a unique heterogenous swine population for m^(6)A sequencing(m^(6)A-seq).We discovered 70genes show both differential RNA expression and m^(6)A modification from high and low IMF group,including ADIPOQ and SFRP1,two hub genes inferred through gene co-expression analysis.Particularly,we observed ADIPOQ,which contains three m^(6)A modification sites within 3’untranslated and protein coding region,could promote porcine intramuscular preadipocyte differentiation in an m^(6)A-dependent manner.Furthermore,we found the YT521-B homology domain family protein 1(YTHDF1)could target and promote ADIPOQ mRNA translation.Conclusions Our study provided a comprehensive profiling of m^(6)A methylation in porcine longissimus dorsi muscle and characterized the involvement of m^(6)A epigenetic modification in the regulation of ADIPOQ mRNA on IMF deposition through an m^(6)A-YTHDF1-dependent manner.展开更多
目的探讨m6A甲基化修饰识别蛋白YTH家族蛋白1(YTH domain family protein 1,YTHDF1)促进肝癌细胞肿瘤干性及激活Wnt/β-catenin信号通路促进肝癌进展的机制。方法采用Western印迹法检测6组肝癌组织和癌旁肝组织中YTHDF1蛋白的表达情况...目的探讨m6A甲基化修饰识别蛋白YTH家族蛋白1(YTH domain family protein 1,YTHDF1)促进肝癌细胞肿瘤干性及激活Wnt/β-catenin信号通路促进肝癌进展的机制。方法采用Western印迹法检测6组肝癌组织和癌旁肝组织中YTHDF1蛋白的表达情况。采用Western印迹法、RIP-PCR、qRT-PCR等检测β-catenin及其下游分子的表达。qRT-PCR、成球实验、克隆形成实验、CCK8等检测YTHDF1对肝癌细胞干性的影响。采用Kaplan-Meier生存曲线分析YTHDF1表达与肝癌患者临床预后的关系。结果肝癌组织中YTHDF1的表达水平明显高于癌旁组织(P<0.001);过表达YTHDF1后,β-catenin及其下游分子的表达增加;过表达YTHDF1后,肝癌细胞的肿瘤干性明显上调、增殖能力显著增强;Kaplan-Meier生存曲线表明YTHDF1高表达的肝癌患者预后更差。结论YTHDF1在肝癌组织中高表达,促进了β-catenin及其下游分子的表达,增强了肝癌细胞的干性,对肝癌患者的预后产生了不良影响。展开更多
Severe immune responses regulate the various clinical hepatic injuries, includingautoimmune hepatitis and acute viral hepatitis. N6-methyladenosine (m6A) modification is acrucial regulator of immunity and inflammation...Severe immune responses regulate the various clinical hepatic injuries, includingautoimmune hepatitis and acute viral hepatitis. N6-methyladenosine (m6A) modification is acrucial regulator of immunity and inflammation. However, the precise role of YTHDF1 in Tcell-mediated hepatitis remains incompletely characterized. To address this, we utilizedConcanavalin A (ConA)-induced mouse liver damage as an experimental model for T cell-mediated hepatitis. Our findings found that hepatic YTHDF1 protein rapidly decreased during ConAinduced hepatitis, and YTHDF1-deficient (Ythdf1^(-/-)) mice showed more susceptibility toConA-induced liver injury, along with an intensified inflammatory storm accompanied by aggravated hepatic inflammatory response via ERK and NF-κB pathways. Interestingly, hepatic-specific over-expression or deletion of YTHDF1 exhibited redundancy in ConA-induced liver injury.Validation in bone marrow chimeric mice confirmed the necessity of YTHDF1 in hematopoieticcells for controlling the response to ConA-induced hepatitis. Additionally, our data revealedthat YTHDF1 deletion in macrophages exacerbated the inflammatory response induced by lipopolysaccharide. In summary, our study uncovered that YTHDF1 deficiency exacerbates the immune response in ConA-induced hepatitis by modulating the expression of inflammatorymediators, highlighting the potential of YTHDF1 as a therapeutic target for clinical hepatitis.展开更多
文摘目的:目前尚不清楚m6A甲基化基因在结直肠癌分化中的作用。本研究结合生物信息学分析与实验验证,探讨这些基因在结直肠癌中的表达特征及其潜在机制。方法:利用生物信息学,我们发现YTHDF1、YTHDF2、ALKBH5、WTAP与结直肠癌表达异常显著(p Background: The role of m6A methylation genes in the differentiation of colorectal cancer is currently unclear. This study combines bioinformatics analysis and experimental validation to explore the expression patterns of these genes in colorectal cancer and their potential mechanisms. Methods: Using bioinformatics, we found that YTHDF1, YTHDF2, ALKBH5, and WTAP are significantly aberrantly expressed in colorectal cancer (p < 0.05). In a real-world cohort from Lishui, quantitative PCR (qPCR) was used to measure mRNA expression of these four genes in CRC tissues and adjacent normal tissues. Multivariable logistic regression analysis with SPSS identified YTHDF1 and WTAP mRNA expression levels as being associated with CRC differentiation. Immunohistochemistry (IHC) was performed to examine corresponding protein levels, quantified using ImageJ. Results showed that YTHDF1 protein expression negatively correlated with CRC differentiation, whereas WTAP protein expression positively correlated. Additionally, multivariable Cox regression analysis was conducted to identify potential factors influencing CRC development. Results: In Lishui, YTHDF1 protein expression was negatively associated with CRC differentiation, while WTAP protein expression was positively associated. Age and alcohol consumption were key risk factors for CRC development, while no significant associations were found with sex or smoking. GO enrichment analysis suggests that YTHDF1 and WTAP may regulate the differentiation process of colorectal cancer by participating in mechanisms such as mRNA modification, translation regulation, and RNA stability. For the poorly differentiated CRC group, time to progression rate (TTP) was 100% and overall survival rate (OS) was 0% within three years. Conclusions: In Lishui City, the YTHDF1 and WTAP genes influence the differentiation of colorectal cancer and may serve as important therapeutic targets and biomarkers for the diagnosis of patients with colorectal cancer.
基金supported by the National Natural Science Foundation of China(82371083,82471100,82121003,82271084)Program of Science and Technology International Cooperation Project of Qinghai province(China)(2022-HZ-814)。
文摘N6-methyladenosine(m^(6)A)modification of mRNA is a critical post-transcriptional regulatory mechanism that modulates mRNA metabolism and neuronal function.The m^(6)A reader YTHDF1 has been shown to enhance the translational efficiency of m^(6)A-modified mRNAs in the brain and is essential for learning and memory.However,its role in the mature retina remains unclear.Herein,we report a novel role of Ythdf1 in the maintenance of retinal function using a genetic knockout model.Loss of Ythdf1 resulted in impaired scotopic electroretinogram(ERG)responses and progressive retinal degeneration.Detailed analyses of rod photoreceptors confirmed substantial degenerative changes in the absence of ciliary defects.Single-cell RNA sequencing revealed comprehensive molecular alterations across all retinal cell types in Ythdf1-deficient retinas.Integrative analysis of methylated RNA immunoprecipitation(MeRIP)sequencing and RIP sequencing identified Tulp1 and Dhx38,two inheritable retinal degeneration disease-associated gene homologs,as direct targets of YTHDF1 in the retina.Specifically,YTHDF1 recognized and bound m^(6)A-modified Tulp1 and Dhx38 mRNA at the coding sequence(CDS),enhancing their translational efficiency without altering mRNA levels.Collectively,these findings highlight the essential role of YTHDF1 in preserving visual function and reveal a novel regulatory mechanism of m^(6)A reader proteins in retinal degeneration,identifying potential therapeutic targets for severe retinopathies.
基金supported by funds from the National Natural Science Foundation of China (Grant No.U21A20249)China Postdoctoral Science Foundation (2022 M712794)。
文摘Background Intramuscular fat(IMF)content is a critical indicator of pork quality,and abnormal IMF is also relevant to human disease as well as aging.Although N6-methyladenosine(m^(6)A)RNA modification was recently found to regulate adipogenesis in porcine intramuscular fat,however,the underlying molecular mechanisms was still unclear.Results In this work,we collected 20 longissimus dorsi muscle samples with high(average 3.95%)or low IMF content(average 1.22%)from a unique heterogenous swine population for m^(6)A sequencing(m^(6)A-seq).We discovered 70genes show both differential RNA expression and m^(6)A modification from high and low IMF group,including ADIPOQ and SFRP1,two hub genes inferred through gene co-expression analysis.Particularly,we observed ADIPOQ,which contains three m^(6)A modification sites within 3’untranslated and protein coding region,could promote porcine intramuscular preadipocyte differentiation in an m^(6)A-dependent manner.Furthermore,we found the YT521-B homology domain family protein 1(YTHDF1)could target and promote ADIPOQ mRNA translation.Conclusions Our study provided a comprehensive profiling of m^(6)A methylation in porcine longissimus dorsi muscle and characterized the involvement of m^(6)A epigenetic modification in the regulation of ADIPOQ mRNA on IMF deposition through an m^(6)A-YTHDF1-dependent manner.
文摘目的探讨m6A甲基化修饰识别蛋白YTH家族蛋白1(YTH domain family protein 1,YTHDF1)促进肝癌细胞肿瘤干性及激活Wnt/β-catenin信号通路促进肝癌进展的机制。方法采用Western印迹法检测6组肝癌组织和癌旁肝组织中YTHDF1蛋白的表达情况。采用Western印迹法、RIP-PCR、qRT-PCR等检测β-catenin及其下游分子的表达。qRT-PCR、成球实验、克隆形成实验、CCK8等检测YTHDF1对肝癌细胞干性的影响。采用Kaplan-Meier生存曲线分析YTHDF1表达与肝癌患者临床预后的关系。结果肝癌组织中YTHDF1的表达水平明显高于癌旁组织(P<0.001);过表达YTHDF1后,β-catenin及其下游分子的表达增加;过表达YTHDF1后,肝癌细胞的肿瘤干性明显上调、增殖能力显著增强;Kaplan-Meier生存曲线表明YTHDF1高表达的肝癌患者预后更差。结论YTHDF1在肝癌组织中高表达,促进了β-catenin及其下游分子的表达,增强了肝癌细胞的干性,对肝癌患者的预后产生了不良影响。
基金supported by grants from the National Natural Science Foundation of China(No.81772539,81972238)the Fundamental Research Funds for the Central Universities of China-Xiamen University(No.20720180048).
文摘Severe immune responses regulate the various clinical hepatic injuries, includingautoimmune hepatitis and acute viral hepatitis. N6-methyladenosine (m6A) modification is acrucial regulator of immunity and inflammation. However, the precise role of YTHDF1 in Tcell-mediated hepatitis remains incompletely characterized. To address this, we utilizedConcanavalin A (ConA)-induced mouse liver damage as an experimental model for T cell-mediated hepatitis. Our findings found that hepatic YTHDF1 protein rapidly decreased during ConAinduced hepatitis, and YTHDF1-deficient (Ythdf1^(-/-)) mice showed more susceptibility toConA-induced liver injury, along with an intensified inflammatory storm accompanied by aggravated hepatic inflammatory response via ERK and NF-κB pathways. Interestingly, hepatic-specific over-expression or deletion of YTHDF1 exhibited redundancy in ConA-induced liver injury.Validation in bone marrow chimeric mice confirmed the necessity of YTHDF1 in hematopoieticcells for controlling the response to ConA-induced hepatitis. Additionally, our data revealedthat YTHDF1 deletion in macrophages exacerbated the inflammatory response induced by lipopolysaccharide. In summary, our study uncovered that YTHDF1 deficiency exacerbates the immune response in ConA-induced hepatitis by modulating the expression of inflammatorymediators, highlighting the potential of YTHDF1 as a therapeutic target for clinical hepatitis.
