TWIST1对急性髓系白血病细胞线粒体功能及化疗耐受性的影响及机制分析

doi:10.19586/j.2095-2341.2025.0120

生物技术进展 ›› 2026, Vol. 16 ›› Issue (1): 190-195.DOI: 10.19586/j.2095-2341.2025.0120

TWIST1对急性髓系白血病细胞线粒体功能及化疗耐受性的影响及机制分析

卢奕闻¹^,²(), 丁一祎¹^,², 郭妮妮¹^,², 黄琬玲¹^,², 徐张静雯¹^,², 张沛雯¹^,², 王楠¹^,², 任倩¹^,², 马小彤¹^,²()

^1.中国医学科学院血液病医院（中国医学科学院血液学研究所）北京协和医学院，血液与健康全国重点实验室，国家血液系统疾病临床医学研究中心，细胞生态海河实验室，天津 300020
^2.天津医学健康研究院，天津 301600

收稿日期:2025-09-08 接受日期:2025-10-22 出版日期:2026-01-25 发布日期:2026-02-12
通讯作者: 马小彤
作者简介:卢奕闻 E-mail: luyiwen@ihcams.ac.cn；
基金资助:
国家自然科学基金项目(82570157);国家自然科学基金项目(82270122)

Effects and Mechanisms of TWIST1 on Mitochondrial Function and Chemoresistance in Acute Myeloid Leukemia Cells

Yiwen LU¹^,²(), Yiyi DING¹^,², Nini GUO¹^,², Wanling HUANG¹^,², Jingwen XU-ZHANG¹^,², Peiwen ZHANG¹^,², Nan WANG¹^,², Qian REN¹^,², Xiaotong MA¹^,²()

^1.State Key Laboratory of Experimental Hematology，National Clinical Research Center for Blood Diseases，Haihe Laboratory of Cell Ecosystem，Institute of Hematology & Blood Diseases Hospital，Chinese Academy of Medical Sciences & Peking Union Medical College，Tianjin 300020，China
^2.Tianjin Institutes of Health Science，Tianjin 301600，China

Received:2025-09-08 Accepted:2025-10-22 Online:2026-01-25 Published:2026-02-12
Contact: Xiaotong MA

摘要/Abstract

摘要：

为了探讨转录因子TWIST1对急性髓系白血病（acute myeloid leukemia, AML）细胞自噬的影响，以及是否通过自噬调控线粒体功能和对化疗药物阿糖胞苷的耐受性，构建包含TWIST1 shRNA及scramble对照慢病毒，感染AML细胞。通过Western blot检测敲降TWIST1的AML细胞中LC3-Ⅱ和p62的表达，分析自噬通量；利用TMRM和MitoTracker染色检测敲降TWIST1对细胞线粒体膜电位和质量的影响，Annexin V/7-AAD检测细胞凋亡水平的改变以及AML细胞对阿糖胞苷敏感性的变化；利用自噬激活剂西罗莫司处理感染shTWIST1的细胞，评价对上述作用的干预效果。结果证实，敲降TWIST1可抑制自噬，表现为LC3-Ⅱ表达降低和p62积累，同时出现线粒体功能受损及细胞凋亡水平显著升高（P<0.05）；西罗莫司可部分恢复自噬流量、改善线粒体功能并降低凋亡率。结果表明，敲降TWIST1增强了AML细胞对阿糖胞苷的敏感性，西罗莫司可部分逆转此效应。综上，TWIST1通过调控自噬影响AML细胞线粒体功能和化疗敏感性，研究结果为AML耐药机制及自噬靶向治疗提供理论依据。

关键词: TWIST1, 自噬, 线粒体功能, 化疗耐药, 急性髓系白血病, 西罗莫司, 阿糖胞苷

Abstract:

The aim of this study was to investigate the effects of transcription factor TWIST1 on autophagy in acute myeloid leukemia （AML） cells， as well as whether it regulates mitochondrial function and tolerance to the chemotherapeutic agent cytarabine via autophagy. Lentiviral vectors carrying TWIST1 shRNA and scramble control were constructed and transduced into AML cells. Western blotting was performed to detect the expression levels of LC3-Ⅱ and p62 in TWIST1-knockdown AML cells for analyzing autophagic flux. TMRM and MitoTracker staining were used to determine the effects of TWIST1 knockdown on mitochondrial membrane potential and mass of AML cells. Annexin V/7-AAD staining was applied to assess the changes in cell apoptosis level and the sensitivity of AML cells to cytarabine. Additionally， TWIST1 shRNA-infected cells were treated with sirolimus （an autophagy activator） to evaluate its intervention effect on the aforementioned processes. The results confirmed that TWIST1 knockdown inhibited autophagy， characterized by decreased LC3-Ⅱ expression and p62 accumulation. Concomitantly， impaired mitochondrial function and a significant increase in cell apoptosis were observed （P<0.05）. Sirolimus could partially restore autophagic flux， improve mitochondrial function， and reduce the apoptosis rate. Moreover， TWIST1 knockdown enhanced the sensitivity of AML cells to cytarabine， which could be partially reversed by sirolimus. In conclusion， TWIST1 affects mitochondrial function and chemosensitivity of AML cells by regulating autophagy. These findings provide a theoretical basis for the mechanism of drug resistance in AML and autophagy-targeted therapy.

Key words: TWIST1, autophagy, mitochondrial function, chemoresistance, acute myeloid leukemia, sirolimus, cytarabine

中图分类号:

Q819

卢奕闻, 丁一祎, 郭妮妮, 黄琬玲, 徐张静雯, 张沛雯, 王楠, 任倩, 马小彤. TWIST1对急性髓系白血病细胞线粒体功能及化疗耐受性的影响及机制分析[J]. 生物技术进展, 2026, 16(1): 190-195.

Yiwen LU, Yiyi DING, Nini GUO, Wanling HUANG, Jingwen XU-ZHANG, Peiwen ZHANG, Nan WANG, Qian REN, Xiaotong MA. Effects and Mechanisms of TWIST1 on Mitochondrial Function and Chemoresistance in Acute Myeloid Leukemia Cells[J]. Current Biotechnology, 2026, 16(1): 190-195.

图/表 3

图1 敲降TWIST1对KG-1a细胞自噬相关蛋白表达的影响A：KG-1a细胞中自噬相关蛋白LC3、p62的表达水平；B：LC3的相对表达量；C：p62的相对表达量。 *、**分别表示在P<0.05和P<0.01水平上差异具有统计学意义。

Fig. 1 The effect of TWIST1 knockdown on the expression of autophagy related proteins in KG-1a cells

图2 敲降TWIST1通过抑制自噬诱导线粒体功能障碍与细胞凋亡注：*、**、***分别表示在P<0.05、P<0.01、P<0.001水平上差异具有统计学意义。

Fig. 2 TWIST1 knockdown induces mitochondrial dysfunction and apoptosis by inhibiting autophagy

图3 敲降TWIST1通过调控自噬改变AML细胞对Ara-C的敏感性注：Molm-13细胞经Ara-C联合西罗莫司处理的凋亡率。*表示在P<0.05水平上差异具有统计学意义。

Fig. 3 TWIST1 knockdown alters the sensitivity of AML cells to Ara-C by regulating autophagy

参考文献 22

[1]	DOHNER H, WEISDORF D J, BLOOMFIELD C D. Acute myeloid leukemia[J]. N. Engl. J. Med., 2015, 373(12): 1136-1152.
[2]	欧阳敏,张亚运,刘建新,等.134例老年急性髓系白血病的临床特点及预后分析[J].中国肿瘤临床,2023,50(24):1265-1270.
	OUYANG M, ZHANG Y Y, LIU J X, et al.. Clinical features and prognosis of 134 elderly patients with acute myeloid leukemia[J]. Chin. J. Clin. Oncol., 2023, 50(24): 1265-1270.
[3]	DÖHNER H, WEI A H, LÖWENBERG B. Towards precision medicine for AML[J]. Nat. Rev. Clin. Oncol., 2021, 18(9): 577-590.
[4]	KLIONSKY D J, ABDEL-AZIZ A K, ABDELFATAH S, et al.. Guidelines for the use and interpretation of assays for monitoring autophagy (4th edition)(1)[J]. Autophagy, 2021, 17(1): 445-544.
[5]	DEBNATH J, GAMMOH N, RYAN K M. Autophagy and autophagy-related pathways in cancer[J]. Nat. Rev. Mol. Cell Biol., 2023, 24(8): 560-575.
[6]	RUSSELL R C, GUAN K L. The multifaceted role of autophagy in cancer[J/OL]. EMBO J., 2022, 41(13): e110031[2025-07-14]. .
[7]	AMARAVADI R K, KIMMELMAN A C, DEBNATH J. Targeting autophagy in cancer: recent advances and future directions[J]. Cancer Discov., 2019, 9(9): 1167-1181.
[8]	SEO W, SILWAL P, SONG I C, et al.. The dual role of autophagy in acute myeloid leukemia[J/OL]. J. Hematol. Oncol., 2022, 15(1): 51[2025-09-10]. .
[9]	DU W, XU A, HUANG Y, et al.. The role of autophagy in targeted therapy for acute myeloid leukemia[J]. Autophagy, 2021, 17(10): 2665-2679.
[10]	WHITE E. Deconvoluting the context-dependent role for autophagy in cancer[J]. Nat. Rev. Cancer, 2012, 12(6): 401-410.
[11]	FÜLLGRABE J, KLIONSKY D J, JOSEPH B. The return of the nucleus: transcriptional and epigenetic control of autophagy[J]. Nat. Rev. Mol. Cell Biol., 2014, 15(1): 65-74.
[12]	YANG J, MANI S A, DONAHER J L, et al.. Twist, a master regulator of morphogenesis, plays an essential role in tumor metastasis[J]. Cell, 2004, 117(7): 927-939.
[13]	PUISIEUX A, BRABLETZ T, CARAMEL J. Oncogenic roles of EMT-inducing transcription factors[J]. Nat. Cell Biol., 2014, 16(6): 488-494.
[14]	YU X, HE T, TONG Z, et al.. Molecular mechanisms of TWIST1-regulated transcription in EMT and cancer metastasis[J/OL]. EMBO Rep., 2023, 24(11): e56902[2025-7-14]. .
[15]	WANG N, GUO D, ZHAO Y Y, et al.. TWIST-1 promotes cell growth, drug resistance and progenitor clonogenic capacities in myeloid leukemia and is a novel poor prognostic factor in acute myeloid leukemia[J]. Oncotarget, 2015, 6(25): 20977-20992.
[16]	SHI Y X, SUN Z W, JIA D L, et al.. Autophagy deficiency promotes lung metastasis of prostate cancer via stabilization of TWIST1[J]. Clin. Transl. Oncol., 2022, 24(7): 1403-1412.
[17]	SCHLÄFLI A M, ADAMS O, GALVÁN J A, et al.. Prognostic value of the autophagy markers LC3 and p62/SQSTM1 in early-stage non-small cell lung cancer[J]. Oncotarget, 2016, 7(26): 39544-39555.
[18]	MIZUSHIMA N, YOSHIMORI T. How to interpret LC3 immunoblotting[J]. Autophagy, 2007, 3(6): 542-545.
[19]	HUANG X, ZHANG J, YAO J, et al.. Phase separation of p62: roles and regulations in autophagy[J]. Trends Cell Biol., 2025, 35(10): 854-865.
[20]	WANG N, YIN J, YOU N, et al.. Twist family BHLH transcription factor 1 is required for the maintenance of leukemia stem cell in MLL-AF9(+) acute myeloid leukemia[J]. Haematologica, 2024, 109(1): 84-97.
[21]	朱丽娜,宋志灵.自噬与细胞凋亡:相互作用及其在疾病中的作用[J].生物技术进展,2025,15(4):622-626.
	ZHU L N, SONG Z L. Autophagy and apoptosis: interactions and their role in disease[J]. Curr. Biotechnol., 2025, 15(4): 622-626.
[22]	QIANG L, HE Y Y. Autophagy deficiency stabilizes TWIST1 to promote epithelial-mesenchymal transition[J]. Autophagy, 2014, 10(10): 1864-1865.

TWIST1对急性髓系白血病细胞线粒体功能及化疗耐受性的影响及机制分析

Effects and Mechanisms of TWIST1 on Mitochondrial Function and Chemoresistance in Acute Myeloid Leukemia Cells

RichHTML

PDF

可视化

摘要/Abstract

引用本文

使用本文

图/表 3

参考文献 22

相关文章 15

编辑推荐

Metrics

本文评价

[1]	郭利佩, 张群昌, 曹馨, 张秀玲. CPEB1通过抑制上皮间质转化逆转卵巢癌细胞化疗耐药的研究[J]. 生物技术进展, 2026, 16(1): 205-212.
[2]	江策, 刘昳, 赵亚磊, 常香荣. miR-5589-5p靶向调控FOXK1对肝癌细胞凋亡和自噬的影响[J]. 生物技术进展, 2025, 15(6): 1086-1093.
[3]	袁佳佳, 邵欢. KLF1在急性髓系白血病中的表达及临床意义研究[J]. 生物技术进展, 2025, 15(5): 904-912.
[4]	朱丽娜, 宋志灵. 自噬与细胞凋亡：相互作用及其在疾病中的作用[J]. 生物技术进展, 2025, 15(4): 622-626.
[5]	王彦杰, 邱波. P53基因在骨肉瘤中的作用与研究进展[J]. 生物技术进展, 2025, 15(2): 241-246.
[6]	李鸿博, 陈朱玥, 吕新星. 亚精胺缓解细胞衰老及衰老相关疾病的研究进展[J]. 生物技术进展, 2024, 14(3): 388-398.
[7]	刘一鸣, 谢乐灵, 邢海燕, 唐克晶, 王敏, 饶青. 靶向CD74的嵌合抗原受体T细胞的抗白血病细胞作用研究[J]. 生物技术进展, 2024, 14(3): 501-505.
[8]	潘舟, 胡克. 线粒体功能障碍在缺氧性肺动脉高压中的作用[J]. 生物技术进展, 2023, 13(6): 882-888.
[9]	李玉珍, 朱杰夫, 吴雄飞. PIM1激酶在顺铂诱导的急性肾损伤中的作用[J]. 生物技术进展, 2023, 13(2): 298-304.
[10]	黄琬玲, 朱文琦, 郭妮妮, 王楠, 任倩, 马小彤. NRG4基因对急性髓系白血病细胞增殖、凋亡及周期的影响[J]. 生物技术进展, 2023, 13(2): 305-310.
[11]	张月明, 罗雅琴, 郑伟, 徐杰, 董雪燕, 李善琛, 王敬毅. 急性髓系白血病mRNA与非编码RNA差异表达谱及CeRNA调控网络分析[J]. 生物技术进展, 2023, 13(1): 146-153.
[12]	赵云鹏, 张浩林, 熊倩倩, 李雨婷, 王娟. COPII囊泡衣被蛋白SEC24A在巨自噬通路中的功能研究[J]. 生物技术进展, 2022, 12(6): 906-914.
[13]	侯凯耀, 张二飞, 郑李娜, 陈红光, 谢克亮. 富氢液对脓毒症小鼠心肌细胞线粒体自噬的影响[J]. 生物技术进展, 2022, 12(4): 497-502.
[14]	谢月樵, 郭丹, 王楠, 尤娜, 黄琬玲, 马小彤. TWIST1在人脐静脉内皮细胞增殖、迁移及血管形成中的作用[J]. 生物技术进展, 2022, 12(1): 142-148.
[15]	王珍,杨洛,廖敏,郝亚荣. mTOR信号通路在糖尿病肾病发病机制中的研究进展[J]. 生物技术进展, 2021, 11(3): 316-321.