Overcoming multi-drug resistance in SCLC: a synergistic approach with venetoclax and hydroxychloroquine targeting the lncRNA LYPLAL1-DT/BCL2/BECN1 pathway.

IF 27.7 1区 医学 Q1 BIOCHEMISTRY & MOLECULAR BIOLOGY
Shuxin Li, Jianyi Lv, Zhihui Li, Qiuyu Zhang, Jing Lu, Xueyun Huo, Meng Guo, Xin Liu, Changlong Li, Jinghui Wang, Hanping Shi, Li Deng, Zhenwen Chen, Xiaoyan Du
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引用次数: 0

Abstract

Background: Small cell lung cancer (SCLC) stands as one of the most lethal malignancies, characterized by a grim diagnosis and prognosis. The emergence of multi-drug resistance poses a significant hurdle to effective therapy. Although previous studies have implicated the long noncoding RNA LYPLAL1-DT in the tumorigenesis of SCLC, the precise role of the highly expressed LYPLAL1-DT in SCLC chemoresistance and the underlying mechanism remain inadequately understood.

Methods: cDDP-, VP-16- and PTX-resistant SCLC cells lines were established. The viabilities of SCLC cells were assessed by CCK-8 assay in vitro and xenograft tumor formation assay in vivo. Apoptosis was evaluated by FACS, Western blot and JC-1 fluorescence staining, while autophagy was explored via autophagic flux detection under confocal microscopy and autophagic vacuole investigation under transmission electron microscopy (TEM). The functional role and mechanism of LYPLAL1-DT were further investigated by gain- and loss-of-function assays in vitro. Furthermore, the therapeutic efficacy of the combination of venetoclax and HCQ with cDDP, VP-16 or PTX was evaluated by cell line, cell-derived xenograft (CDX) and patient-derived xenograft (PDX) mice model.

Results: Our findings revealed that LYPLAL1-DT is upregulated in chemoresistant SCLC cell lines. Gain- and loss-of-function assays demonstrated that LYPLAL1-DT impairs sensitivity to cDDP, VP-16, or PTX both in vitro and in vivo. Overexpression of LYPLAL1-DT significantly enhanced autophagy and inhibited apoptosis in SCLC cells. Further analyses, including RIP and RNA pull-down assays, revealed that LYPLAL1-DT promotes the expression of BCL2 by sponging miR-204-5p and is implicated in the assembly of the autophagy-specific complex (BECN1/PtdIns3K complex). Combining venetoclax and HCQ with cDDP, VP-16, or PTX effectively mitigated chemoresistance in SCLC cells and suppressed tumor growth in CDX and PDX models without inducing obvious toxic effects.

Conclusions: Our findings demonstrate that upregulation of LYPLAL1-DT sequesters apoptosis through the LYPLAL1-DT/miR-204-5p/BCL2 axis and promotes autophagy by facilitating the assembly of the BECN1/PtdIns3K complex, thereby mediating multi-drug resistance of SCLC. The triple combination of venetoclax, HCQ, in conjunction with cDDP, VP-16 or PTX overcomes refractory SCLC, shedding light on a potential therapeutic target for combating SCLC chemoresistance.

克服SCLC的多重耐药性:以lncRNA LYPLAL1-DT/BCL2/BECN1通路为靶点的venetoclax和羟氯喹协同方法
背景:小细胞肺癌(SCLC小细胞肺癌(SCLC)是致死率最高的恶性肿瘤之一,其诊断和预后均十分严峻。多种药物耐药性的出现是有效治疗的一大障碍。尽管之前的研究表明长非编码 RNA LYPLAL1-DT 与 SCLC 的肿瘤发生有关,但高表达的 LYPLAL1-DT 在 SCLC 化疗耐药中的确切作用及其内在机制仍未得到充分了解。方法:建立了 cDDP-、VP-16- 和 PTX 抗性 SCLC 细胞系,通过 CCK-8 检测体外 SCLC 细胞的存活率和异种移植肿瘤形成检测体内 SCLC 细胞的存活率。细胞凋亡通过 FACS、Western 印迹和 JC-1 荧光染色进行评估,自噬则通过共聚焦显微镜下的自噬通量检测和透射电子显微镜(TEM)下的自噬空泡研究进行探讨。通过体外功能增益和功能缺失实验进一步研究了LYPLAL1-DT的功能作用和机制。此外,我们还通过细胞系、细胞衍生异种移植(CDX)和患者衍生异种移植(PDX)小鼠模型评估了venetoclax和HCQ与cDDP、VP-16或PTX联合治疗的疗效:我们的研究结果表明,LYPLAL1-DT在化疗耐药的SCLC细胞系中上调。功能增益和功能缺失试验表明,LYPLAL1-DT在体外和体内都会降低对cDDP、VP-16或PTX的敏感性。过表达 LYPLAL1-DT 能显著增强 SCLC 细胞的自噬作用并抑制细胞凋亡。进一步的分析(包括 RIP 和 RNA 下拉实验)显示,LYPLAL1-DT 通过疏导 miR-204-5p 促进 BCL2 的表达,并与自噬特异性复合物(BECN1/PtdIns3K 复合物)的组装有关。将venetoclax和HCQ与cDDP、VP-16或PTX联合使用,可有效减轻SCLC细胞的化疗耐药性,并抑制CDX和PDX模型中的肿瘤生长,且不会引起明显的毒性反应:我们的研究结果表明,LYPLAL1-DT的上调可通过LYPLAL1-DT/miR-204-5p/BCL2轴抑制细胞凋亡,并通过促进BECN1/PtdIns3K复合物的组装来促进自噬,从而介导SCLC的多重耐药。venetoclax、HCQ与cDDP、VP-16或PTX的三联疗法克服了难治性SCLC,揭示了对抗SCLC化疗耐药性的潜在治疗靶点。
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来源期刊
Molecular Cancer
Molecular Cancer 医学-生化与分子生物学
CiteScore
54.90
自引率
2.70%
发文量
224
审稿时长
2 months
期刊介绍: Molecular Cancer is a platform that encourages the exchange of ideas and discoveries in the field of cancer research, particularly focusing on the molecular aspects. Our goal is to facilitate discussions and provide insights into various areas of cancer and related biomedical science. We welcome articles from basic, translational, and clinical research that contribute to the advancement of understanding, prevention, diagnosis, and treatment of cancer. The scope of topics covered in Molecular Cancer is diverse and inclusive. These include, but are not limited to, cell and tumor biology, angiogenesis, utilizing animal models, understanding metastasis, exploring cancer antigens and the immune response, investigating cellular signaling and molecular biology, examining epidemiology, genetic and molecular profiling of cancer, identifying molecular targets, studying cancer stem cells, exploring DNA damage and repair mechanisms, analyzing cell cycle regulation, investigating apoptosis, exploring molecular virology, and evaluating vaccine and antibody-based cancer therapies. Molecular Cancer serves as an important platform for sharing exciting discoveries in cancer-related research. It offers an unparalleled opportunity to communicate information to both specialists and the general public. The online presence of Molecular Cancer enables immediate publication of accepted articles and facilitates the presentation of large datasets and supplementary information. This ensures that new research is efficiently and rapidly disseminated to the scientific community.
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