背景:小细胞外囊泡(sEV)介导肿瘤微环境(TME)中的细胞间通讯,并有助于肿瘤的恶性转化。包括不受限制的增长,转移,或治疗抗性。然而,目前缺乏靶向sEVs的药物,通过sEVs介导的TME重编程克服或逆转肿瘤化疗耐药.方法:采用紫杉醇(PTX)耐药的A549T细胞系,探讨α-hederin通过小细胞外囊泡(sEVs)途径对非小细胞肺癌(NSCLC)化疗耐药传递的抑制作用。该研究利用CCK-8测定和流式细胞术。转录组学,蛋白质印迹,油红O染色,和靶向代谢组学用于评估α-hederin对治疗前后NSCLC细胞中化疗耐药传递相关信号通路表达的影响。进行体内分子成像和免疫组织化学以评估α-hederin如何通过sEV途径影响化学抗性的传播。采用RT-PCR来检测响应于α-hederin处理的miRNA和lncRNA的表达。结果:α-hederin通过对sEV分泌的依赖性克服了A549T细胞对PTX化疗的耐药性。然而,当囊泡分泌被GW4869抑制剂阻断时,α-hederin的有效性受到损害.暴露于A549T衍生的sEV的受体细胞中463个上调基因的转录组学分析显示,这些sEV增强了TGFβ信号传导和不饱和脂肪酸合成途径。α-hederin通过降低sEVs介导的TGFβ/SMAD2通路的信号活性来抑制15种不饱和脂肪酸合成。Further,我们观察到α-hederin促进了三种microRNAs(miRNAs,包括miR-21-5p,miR-23a-3p,和miR-125b-5p),并在A549T细胞中分选为sEV。这些miRNA靶向sEV受体细胞中的TGFβ/SMAD信号传导活性,并使其对PTX疗法敏感。结论:我们的发现表明,α-hederin可以通过sEV介导的多个miRNA积累来敏感PTX耐药的NSCLC细胞。并抑制受体细胞中的TGFβ/SMAD2通路。
Background: Small extracellular vesicles (sEVs) mediate intercellular communication in the tumor microenvironment (TME) and contribute to the malignant transformation of tumors, including unrestricted growth, metastasis, or therapeutic resistance. However, there is a lack of agents targeting sEVs to overcome or reverse tumor chemotherapy resistance through sEVs-mediated TME reprogramming. Methods: The paclitaxel (PTX)-resistant A549T cell line was used to explore the inhibitory effect of alpha-hederin on impeding the transmission of chemoresistance in non-small cell lung cancer (NSCLC) through the small extracellular vesicles (sEVs) pathway. This investigation utilized the CCK-8 assay and flow cytometry. Transcriptomics, Western blot, oil red O staining, and targeted metabolomics were utilized to evaluate the impact of alpha-hederin on the expression of signaling pathways associated with chemoresistance transmission in NSCLC cells before and after treatment. In vivo molecular imaging and immunohistochemistry were conducted to assess how alpha-hederin influences the transmission of chemoresistance through the sEVs pathway. RT-PCR was employed to examine the expression of miRNA and lncRNA in response to alpha-hederin treatment. Results: The resistance to PTX chemotherapy in A549T cells was overcome by alpha-hederin through its dependence on sEV secretion. However, the effectiveness of alpha-hederin was compromised when vesicle secretion was blocked by the GW4869 inhibitor. Transcriptomic analysis for 463 upregulated genes in recipient cells exposed to A549T-derived sEVs revealed that these sEVs enhanced TGFβ signaling and unsaturated fatty acid synthesis pathways. Alpha-hederin inhibited 15 types of unsaturated fatty acid synthesis by reducing the signaling activity of the sEVs-mediated TGFβ/SMAD2 pathway. Further, we observed that alpha-hederin promoted the production of three microRNAs (miRNAs, including miR-21-5p, miR-23a-3p, and miR-125b-5p) and the sorting to sEVs in A549T cells. These miRNAs targeted the TGFβ/SMADs signaling activity in sEVs-recipient cells and sensitized them to the PTX therapy. Conclusion: Our finding demonstrated that alpha-hederin could sensitize PTX-resistant NSCLC cells by sEV-mediated multiple miRNAs accumulation, and inhibiting TGFβ/SMAD2 pathways in recipient cells.