Blockage of transferred exosome-shuttled miR-494 inhibits melanoma growth and metastasis
Blockage of transferred exosome-shuttled miR-494 inhibits melanoma growth and metastasis
复制标题
阻断转移的外泌体穿梭的 miR-494 可抑制黑色素瘤的生长和转移
DOI：
10.1002/jcp.28234
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发表时间：
2019-09-01
影响因子：
5.6
通讯作者：
Zhou, Jianda
中科院分区：
文献类型：
--
作者：
Li, Jingjing;Chen, Jia;Zhou, Jianda
有新的证据表明，黑色素瘤中的生物活性物质可以通过外切体转运。然而，这种癌症进展背后的外体内容的功能仍然很大程度上是未知的。我们的目的是确定外切体分泌是否有助于细胞microRNA-494(miR-494)的丢失，并研究miR-494在黑色素瘤进展中的作用。用超速离心法分离血清和细胞培养条件培养液中的外切体。用短发夹状RNA沉默抑制外切酶的Rab27a。为了解决外体miR-494的功能作用，我们评估了细胞的增殖、迁移、侵袭能力和细胞凋亡。最后，建立了异种皮下移植和肺转移模型，以确定外体miR-494的体内作用。通过对基因表达总表数据库中的黑素细胞和黑色素瘤来源的外切体进行长非编码RNA微阵列分析，我们发现miR-494在黑色素瘤来源的外切体中富含。黑色素瘤患者血清和A375细胞分泌的外切体miR-494表达增加。Rab27a的缺失减少了外切体的分泌，挽救了细胞miR-494的丰度。功能研究表明，Rab27a基因的敲除和miR-494随后的积聚通过诱导细胞凋亡显著抑制了黑色素瘤细胞的恶性表型。裸鼠实验证实，在Rab27a缺失后，miR-494的积聚增加，从而抑制了肿瘤的生长和转移。总之，阻断转运的外切体穿梭miR-494是治疗黑色素瘤的一种潜在选择。
There is emerging evidence of bioactive material transport by exosomes in melanoma. However, the functions of exosome content underlying such cancer progression remain largely unknown. We aimed at determining whether exosome secretion contributes to cellular microRNA-494 (miR-494) loss and investigated the roles of miR-494 in melanoma progression. The exosomes from blood serum and cell culture conditioned media were separated by ultracentrifugation. A short hairpin RNA was used to silence rab27a for inhibiting exosomerelease. To address the functional role of exosomal miR-494, we assessed cell proliferation, migration, invasion capabilities, and cell apoptosis. Finally, subcutaneous xenograft and lung-metastasis models were constructedto determine the effect of exosomal miR-494 in vivo. Based on long noncoding RNA microarray analysis of melanocyte and melanoma-derived exosomes from the Gene Expression Omnibus database, we discovered that miR-494 was enriched in melanoma-derived exosomes. And miR-494 was increased in exosomes secreted from melanoma patients' serum and A375 cells. Rab27a depletion reduced exosome secretion and rescued the abundance of cellular miR-494. Functional studies revealed that knockdown of rab27a and subsequent accumulation of miR-494 significantly suppressed the malignant phenotypes of melanoma cells via inducing cell apoptosis. Nude mice experiments confirmed that tumor growth and metastasis were suppressed by increasing miR-494 accumulation after rab27a depletion. In conclusion, blocking transferred exosome-shuttled miR-494 is a potential therapeutic option for melanoma.