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Published on: September 8, 2017
Alpha-hederin reprograms multi-miRNAs activity and overcome small extracellular vesicles-mediated paclitaxel
Yuzhen Chang1,2, Xinyu Gao3, Yuchen Jiang1
1Shanghai Key Laboratory of Molecular Imaging, Jiading District Central Hospital Affiliated Shanghai University of Medicine and Health Sciences, Shanghai, China.
Abstract:
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.
Insights
Alpha-hederin overcomes paclitaxel resistance in non-small cell lung cancer by targeting small extracellular vesicles (sEVs). This natural compound sensitizes resistant cells by inhibiting TGFβ/SMAD2 signaling and promoting miRNA accumulation within sEVs.
Area of Science:
- Oncology
- Cell Biology
- Molecular Medicine
Background:
- Small extracellular vesicles (sEVs) play a crucial role in tumor microenvironment communication, contributing to cancer progression and therapeutic resistance.
- Targeting sEVs offers a potential strategy to overcome chemoresistance by reprogramming the tumor microenvironment.
- Current therapeutic approaches lack agents specifically designed to reverse tumor chemoresistance mediated by sEVs.
Purpose of the Study:
- To investigate the efficacy of alpha-hederin in overcoming paclitaxel (PTX) resistance in non-small cell lung cancer (NSCLC) via the small extracellular vesicle (sEV) pathway.
- To elucidate the molecular mechanisms by which alpha-hederin modulates chemoresistance transmission through sEVs.
- To evaluate the potential of alpha-hederin as a therapeutic agent for PTX-resistant NSCLC.
Main Methods:
- Utilized a paclitaxel-resistant A549T cell line and GW4869 inhibitor to study sEV secretion.
- Employed CCK-8 assay, flow cytometry, transcriptomics, Western blot, oil red O staining, and metabolomics to analyze cellular and molecular changes.
- Conducted in vivo molecular imaging, immunohistochemistry, and RT-PCR to assess the impact of alpha-hederin on chemoresistance transmission and miRNA/lncRNA expression.
Main Results:
- Alpha-hederin effectively overcame PTX resistance in A549T cells, dependent on sEV secretion.
- Transcriptomic analysis revealed sEVs from resistant cells enhanced TGFβ signaling and unsaturated fatty acid synthesis; alpha-hederin inhibited these pathways.
- Alpha-hederin promoted the sorting of miR-21-5p, miR-23a-3p, and miR-125b-5p into sEVs, which targeted and reduced TGFβ/SMADs signaling in recipient cells, sensitizing them to PTX.
Conclusions:
- Alpha-hederin sensitizes PTX-resistant NSCLC cells by modulating sEV cargo, specifically through the accumulation of multiple miRNAs.
- The mechanism involves the inhibition of TGFβ/SMAD2 pathways in recipient cells via sEV-mediated miRNA delivery.
- Alpha-hederin represents a promising therapeutic strategy for overcoming chemoresistance in NSCLC by targeting sEV-mediated intercellular communication.
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