Chemotherapy is currently the mainstay of systemic management for triple-negative breast cancer(TNBC),but chemoresistance significantly impacts patient outcomes.Our research indicates that Doxorubicin(Dox)-resistant T...Chemotherapy is currently the mainstay of systemic management for triple-negative breast cancer(TNBC),but chemoresistance significantly impacts patient outcomes.Our research indicates that Doxorubicin(Dox)-resistant TNBC cells exhibit increased glycolysis and ATP generation compared to their parental cells,with this metabolic shift contributing to chemoresistance.We discovered that ALKBH3,an m^(1)A demethylase enzyme,is crucial in regulating the enhanced glycolysis in Doxresistant TNBC cells.Knocking down ALKBH3 reduced ATP generation,glucose consumption,and lactate production,implicating its involvement in mediating glycolysis.Further investigation revealed that aldolase A(ALDOA),a key enzyme in glycolysis,is a downstream target of ALKBH3.ALKBH3 regulates ALDOA mRNA stability through m^(1)A demethylation at the 30-untranslated region(30UTR).This methylation negatively affects ALDOA mRNA stability by recruiting the YTHDF2/PAN2ePAN3 complex,leading to mRNA degradation.The ALKBH3/ALDOA axis promotes Dox resistance both in vitro and in vivo.Clinical analysis demonstrated that ALKBH3 and ALDOA are upregulated in breast cancer tissues,and higher expression of these proteins is associated with reduced overall survival in TNBC patients.Our study highlights the role of the ALKBH3/ALDOA axis in contributing to Dox resistance in TNBC cells through regulation of ALDOA mRNA stability and glycolysis.展开更多
Objective:To use bioinformatics analysis and in vitro and in vivo experiments to study the biological role of ALKBH3 in lung adenocarcinoma.Methods:Bioinformatics analysis of ALKBH3 was performed using databases.ALKBH...Objective:To use bioinformatics analysis and in vitro and in vivo experiments to study the biological role of ALKBH3 in lung adenocarcinoma.Methods:Bioinformatics analysis of ALKBH3 was performed using databases.ALKBH3 expression in lung adenocarcinoma and adjacent tissues was detected by qPCR(quantitative polymerase chain reaction),western blotting,and immunohistochemistry.Stable transformed A549 cells with low expression of ALKBH3 were constructed.The effects of knockdown of ALKBH3 on the proliferation,migration,and invasion of lung adenocarcinoma A549 cells were detected by CCK-8,cell scratch,and transwell invasion assays,respectively.The effects of ALKBH3 on the proliferation of A549 cells in vivo were detected using subcutaneous tumorigenesis in nude mice.Results:Bioinformatics analysis showed that ALKBH3 has diagnostic value in tumors such as lung adenocarcinoma,the expression of ALKBH3 is related to immune cell infiltration,ALKBH3 interacts with ASCC family molecules,and ALKBH3 is involved in the demethylation of DNA and RNA.The expression of ALKBH3 in lung adenocarcinoma was higher than that in adjacent tissues(P<0.05).CCK-8,wound healing and transwell assays showed that ALKBH3 knockdown significantly inhibited the proliferation,migration,and invasion of A549 cells in vitro(P<0.01).ALKBH3 knockdown also significantly inhibited the growth of subcutaneous tumors in nude mice(P<0.01).Conclusions:ALKBH3 is a potential diagnostic marker for lung adenocarcinoma.Results in vivo and in vitro showed that knocking down ALKBH3 could inhibit the proliferation,migration,invasion,and subcutaneous tumorigenesis of lung adenocarcinoma A549 cells.展开更多
ALKBH3,a demethylase responsible for demethylating N1-methyladenosine(m^(1)A)in mRNA and N1-methyldeoxyadenosine in single-stranded DNA,plays an important role in DNA repair and cancer cell proliferation.However,its f...ALKBH3,a demethylase responsible for demethylating N1-methyladenosine(m^(1)A)in mRNA and N1-methyldeoxyadenosine in single-stranded DNA,plays an important role in DNA repair and cancer cell proliferation.However,its function in hematopoietic stem cells(HSCs)is unknown.In this study,we generated Alkbh3 knockout mice and observed that the deletion of Alkbh3 does not impair the reconstitution capacity of HSCs in both primary and secondary transplantation.Aged hematopoietic stem and progenitor cells exhibit increased expression of ALKBH3.Forced ALKBH3 rescued the differentiation skewing without affecting the reconstitution capacity of aged HSCs.In brief,our study for the first time investigated the functional role of ALKBH3 in hematopoietic system,and observed that ALKBH3 is dispensable for HSCs maintenance and differentiation,but overexpression of ALKBH3 rectified the differentiation skewing of aged HSCs.展开更多
胃癌是消化系统最常见的恶性肿瘤之一,多数患者发现时已处于晚期,预后不佳。外科手术及化学治疗(化疗仍是目前胃癌的主要治疗方式。N6-甲基腺嘌呤(N6-methyladenosine,m^(6)A)是近年来肿瘤研究的热点。m^(6)A作为真核生物中最常见的RNA...胃癌是消化系统最常见的恶性肿瘤之一,多数患者发现时已处于晚期,预后不佳。外科手术及化学治疗(化疗仍是目前胃癌的主要治疗方式。N6-甲基腺嘌呤(N6-methyladenosine,m^(6)A)是近年来肿瘤研究的热点。m^(6)A作为真核生物中最常见的RNA修饰形式,可以调控RNA循环的各个阶段,包括RNA剪接、加工、降解和翻译等,从而调控RNA的表达和功能,在细胞分化、发育和代谢等各个环节中发挥关键作用。m^(6)A去甲基化酶可去除RNA上的甲基基团,确保m^(6)A甲基化是一个动态的可逆的过程。作为m^(6)A甲基化过程的关键酶,m^(6)A去甲基化酶——脂肪和肥胖相关蛋白(fat mass and obesity-associated protein,FTO)、AlkB同系物5(AlkB homolog 5,ALKBH5)、ALKBH3的失调能通过多种机制调控胃癌的演进过程,与胃癌的发生发展密切相关。m^(6)A去甲基化酶通过调节信号通路,改变胃癌细胞的增殖和侵袭能力,影响胃癌对化疗药物的耐药性,参与调控胃癌的免疫应答及线粒体代谢,从而影响胃癌细胞的生长,有望成为一个全新的治疗靶点。该文综述了m^(6)A去甲基化酶参与胃癌发生发展的分子机制,以及其表达和功能与胃癌生物学特性的关系,旨在为胃癌的早期诊断和靶向治疗提供新的研究思路。展开更多
基金funded by the Research Project of TCM Bureau of Guangdong Province(20231324,China)the Special Fund of Foshan Climbing Peak Plan(2020B018,China)+1 种基金the Basic and Applied Basic Research Foundation of Guangdong Province(Grant No.2022A1515140091,China)the Natural Science Foundation of Guangdong Province(2023A1515030291,China).
文摘Chemotherapy is currently the mainstay of systemic management for triple-negative breast cancer(TNBC),but chemoresistance significantly impacts patient outcomes.Our research indicates that Doxorubicin(Dox)-resistant TNBC cells exhibit increased glycolysis and ATP generation compared to their parental cells,with this metabolic shift contributing to chemoresistance.We discovered that ALKBH3,an m^(1)A demethylase enzyme,is crucial in regulating the enhanced glycolysis in Doxresistant TNBC cells.Knocking down ALKBH3 reduced ATP generation,glucose consumption,and lactate production,implicating its involvement in mediating glycolysis.Further investigation revealed that aldolase A(ALDOA),a key enzyme in glycolysis,is a downstream target of ALKBH3.ALKBH3 regulates ALDOA mRNA stability through m^(1)A demethylation at the 30-untranslated region(30UTR).This methylation negatively affects ALDOA mRNA stability by recruiting the YTHDF2/PAN2ePAN3 complex,leading to mRNA degradation.The ALKBH3/ALDOA axis promotes Dox resistance both in vitro and in vivo.Clinical analysis demonstrated that ALKBH3 and ALDOA are upregulated in breast cancer tissues,and higher expression of these proteins is associated with reduced overall survival in TNBC patients.Our study highlights the role of the ALKBH3/ALDOA axis in contributing to Dox resistance in TNBC cells through regulation of ALDOA mRNA stability and glycolysis.
基金funded by the Sichuan Provincial Health Commission Project Fund(No.21PJ188).
文摘Objective:To use bioinformatics analysis and in vitro and in vivo experiments to study the biological role of ALKBH3 in lung adenocarcinoma.Methods:Bioinformatics analysis of ALKBH3 was performed using databases.ALKBH3 expression in lung adenocarcinoma and adjacent tissues was detected by qPCR(quantitative polymerase chain reaction),western blotting,and immunohistochemistry.Stable transformed A549 cells with low expression of ALKBH3 were constructed.The effects of knockdown of ALKBH3 on the proliferation,migration,and invasion of lung adenocarcinoma A549 cells were detected by CCK-8,cell scratch,and transwell invasion assays,respectively.The effects of ALKBH3 on the proliferation of A549 cells in vivo were detected using subcutaneous tumorigenesis in nude mice.Results:Bioinformatics analysis showed that ALKBH3 has diagnostic value in tumors such as lung adenocarcinoma,the expression of ALKBH3 is related to immune cell infiltration,ALKBH3 interacts with ASCC family molecules,and ALKBH3 is involved in the demethylation of DNA and RNA.The expression of ALKBH3 in lung adenocarcinoma was higher than that in adjacent tissues(P<0.05).CCK-8,wound healing and transwell assays showed that ALKBH3 knockdown significantly inhibited the proliferation,migration,and invasion of A549 cells in vitro(P<0.01).ALKBH3 knockdown also significantly inhibited the growth of subcutaneous tumors in nude mice(P<0.01).Conclusions:ALKBH3 is a potential diagnostic marker for lung adenocarcinoma.Results in vivo and in vitro showed that knocking down ALKBH3 could inhibit the proliferation,migration,invasion,and subcutaneous tumorigenesis of lung adenocarcinoma A549 cells.
基金This work was supported by grant numbers 2018YFA0800200,2017YFA0104000,Z181100001818005 and 91849106 to J.WW.,from the National KeyR&D Program of China or the Beijing Municipal Science&Technology Commission and the National Natural Science Foundation of China.
文摘ALKBH3,a demethylase responsible for demethylating N1-methyladenosine(m^(1)A)in mRNA and N1-methyldeoxyadenosine in single-stranded DNA,plays an important role in DNA repair and cancer cell proliferation.However,its function in hematopoietic stem cells(HSCs)is unknown.In this study,we generated Alkbh3 knockout mice and observed that the deletion of Alkbh3 does not impair the reconstitution capacity of HSCs in both primary and secondary transplantation.Aged hematopoietic stem and progenitor cells exhibit increased expression of ALKBH3.Forced ALKBH3 rescued the differentiation skewing without affecting the reconstitution capacity of aged HSCs.In brief,our study for the first time investigated the functional role of ALKBH3 in hematopoietic system,and observed that ALKBH3 is dispensable for HSCs maintenance and differentiation,but overexpression of ALKBH3 rectified the differentiation skewing of aged HSCs.
文摘胃癌是消化系统最常见的恶性肿瘤之一,多数患者发现时已处于晚期,预后不佳。外科手术及化学治疗(化疗仍是目前胃癌的主要治疗方式。N6-甲基腺嘌呤(N6-methyladenosine,m^(6)A)是近年来肿瘤研究的热点。m^(6)A作为真核生物中最常见的RNA修饰形式,可以调控RNA循环的各个阶段,包括RNA剪接、加工、降解和翻译等,从而调控RNA的表达和功能,在细胞分化、发育和代谢等各个环节中发挥关键作用。m^(6)A去甲基化酶可去除RNA上的甲基基团,确保m^(6)A甲基化是一个动态的可逆的过程。作为m^(6)A甲基化过程的关键酶,m^(6)A去甲基化酶——脂肪和肥胖相关蛋白(fat mass and obesity-associated protein,FTO)、AlkB同系物5(AlkB homolog 5,ALKBH5)、ALKBH3的失调能通过多种机制调控胃癌的演进过程,与胃癌的发生发展密切相关。m^(6)A去甲基化酶通过调节信号通路,改变胃癌细胞的增殖和侵袭能力,影响胃癌对化疗药物的耐药性,参与调控胃癌的免疫应答及线粒体代谢,从而影响胃癌细胞的生长,有望成为一个全新的治疗靶点。该文综述了m^(6)A去甲基化酶参与胃癌发生发展的分子机制,以及其表达和功能与胃癌生物学特性的关系,旨在为胃癌的早期诊断和靶向治疗提供新的研究思路。
