Medical Genetics, Faculty of Medicine, Pamukkale University, Denizli, Turkey.
Department of Cancer Molecular Biology, Institute of Medical Sciences, Pamukkale University, Denizli, Turkey.
Mol Biol Rep. 2022 Aug;49(8):7953-7965. doi: 10.1007/s11033-022-07632-6. Epub 2022 Jun 12.
MYC genes are amplified/overexpressed in 20% of SCLCs, showing that Myc and Myc-dependent cellular mechanisms are strong candidates as therapeutic targets in SCLC. Small extracellular vesicles support the carcinogenesis process by acting as messengers delivering nucleic acids and proteins-moreover, no reports associate Myc and the functional effect of small extracellular vesicles in small cell lung cancer.
After the effects of small extracellular vesicles (sEVs) obtained from H82 and H209 cells on HUVEC and MRC-5 cells were observed, the Myc-dependent effect of the sEVs on oncogenic potentials was further evaluated by manipulating Myc expression via lentiviral vectors in H82 and H209 cells. Then, small extracellular vesicles of Myc-manipulated SCLC cells were isolated using sEVs isolation reagents. Finally, HUVEC and MRC5 cells were treated with SCLC-derived small extracellular vesicles. Cellular activity of recipient normal lung cells was investigated by cell growth assay, wound healing assay, and transwell assay. miRNA composition changes in small extracellular vesicles and SCLC cells were investigated using miRNA microarray and QRT-PCR assay. Our results indicated that normal lung cells treated with SCLC-derived small extracellular vesicles had higher proliferation, migration capability than non-treated counterparts. Additionally, after investigating the potential effects of small extracellular vesicles derived from Myc-dysregulated SCLC cell lines, we further evaluated the Myc-dependent miRNA composition in the small extracellular vesicles. The present study revealed that Myc regulates hsa-miR-7, hsa-miR-9, hsa-miR-125b, hsa-miR-181a_2, hsa-miR-455, hsa-miR-642, and hsa-miR-4417 expressions in SCLC cell lines, not only in cellular but also in exosomal content.
Small extracellular vesicles and MYC are essential targets for therapeutic strategy in SCLC. Our study revealed that the expression level of MYC can affect the function of sEVs and encapsulate the miRNA composition in SCLC. Besides, small extracellular vesicles derived from SCLC cells can modulate normal lung cells.
MYC 基因在 20%的 SCLC 中扩增/过表达，表明 Myc 和依赖 Myc 的细胞机制是 SCLC 治疗靶点的有力候选物。小细胞外囊泡通过充当传递核酸和蛋白质的信使来支持癌变过程-此外，尚无报告将 Myc 与小细胞肺癌中小细胞外囊泡的功能效应联系起来。
观察到来自 H82 和 H209 细胞的小细胞外囊泡（sEVs）对 HUVEC 和 MRC-5 细胞的影响后，通过慢病毒载体操纵 Myc 表达，进一步评估 sEVs 对致癌潜力的 Myc 依赖性效应在 H82 和 H209 细胞中。然后，使用 sEV 分离试剂分离 Myc 操纵的 SCLC 细胞的小细胞外囊泡。最后，用 SCLC 来源的小细胞外囊泡处理 HUVEC 和 MRC5 细胞。通过细胞生长测定、划痕愈合测定和 Transwell 测定研究受体正常肺细胞的细胞活性。使用 miRNA 微阵列和 QRT-PCR 测定研究小细胞外囊泡和 SCLC 细胞中小 miRNA 组成的变化。我们的结果表明，用 SCLC 来源的小细胞外囊泡处理的正常肺细胞比未处理的细胞具有更高的增殖和迁移能力。此外，在研究 Myc 失调的 SCLC 细胞系衍生的小细胞外囊泡的潜在作用后，我们进一步评估了小细胞外囊泡中的 Myc 依赖性 miRNA 组成。本研究表明，Myc 调节 SCLC 细胞系中的 hsa-miR-7、hsa-miR-9、hsa-miR-125b、hsa-miR-181a_2、hsa-miR-455、hsa-miR-642 和 hsa-miR-4417 的表达，不仅在细胞内，而且在 exosomal 含量中。
小细胞外囊泡和 MYC 是 SCLC 治疗策略的重要靶点。我们的研究表明，MYC 的表达水平可以影响 sEVs 的功能并包裹 SCLC 中的 miRNA 组成。此外，来自 SCLC 细胞的小细胞外囊泡可以调节正常肺细胞。
