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长链非编码 RNA NEAT1 拮抗重楼苷 B 对黑色素瘤增殖、迁移、侵袭和 EMT 的抑制作用。

LncRNA NEAT1 antagonizes the inhibition of melanoma proliferation, migration, invasion and EMT by Polyphyllin B.

机构信息

Key Laboratory of Xin'an Medicine, Ministry of Education, Anhui University of Chinese Medicine, Meishan Road, Shushan District, Hefei, 230038, China.

School of Pharmacy, Anhui University of Chinese Medicine, Hefei, 230012, China.

出版信息

Naunyn Schmiedebergs Arch Pharmacol. 2023 Oct;396(10):2469-2480. doi: 10.1007/s00210-023-02474-w. Epub 2023 Apr 1.

Abstract

Polyphyllin B (PPB) is a compound with anti-tumor effects. Nuclear paraspeckle assembly transcript 1 (NEAT1) is a long-stranded noncoding RNA that induces epithelial-mesenchymal transition (EMT) of tumor cells and promotes tumor growth and metastasis. However, the role and mechanism of PPB on melanoma and the correlation between them remain unclear. In this study we screened NEAT1 by using LncRNA transcriptomic sequencing, and then transfected B16F10 cells using OVER-NEAT1 lentivirus. Next, we found that PPB had significant proliferation inhibition of melanoma and B16F10 cells through MTT assay and establishment of mouse subcutaneous transplantation tumor model; in addition, through wound healing assay, transwell assay and establishment of mouse melanoma lung metastasis model, we found that PPB significantly inhibited the invasion and migration of B16F10 cells in vitro, and inhibited the metastasis of melanoma to lung, bone and liver in vivo. Finally, changes in the expression levels of EMT-related proteins were assessed by western blot (WB) and immunohistochemistry, and PPB significantly downregulated the expression levels of MMP-9, N-cadherin, etc., and upregulated E-cadherin. While overexpressed NEAT1 showed the ability to promote melanoma proliferation, migration and invasion, in addition to partially reversed the inhibition of proliferation, migration and invasion of melanoma by PPB mentioned above.

摘要

多形棘蝇碱 B(PPB)是一种具有抗肿瘤作用的化合物。核斑 assemble 转录物 1(NEAT1)是一种长链非编码 RNA,可诱导肿瘤细胞上皮-间充质转化(EMT),并促进肿瘤生长和转移。然而,PPB 对黑色素瘤的作用及其机制尚不清楚。在本研究中,我们通过 LncRNA 转录组测序筛选出 NEAT1,然后使用 OVER-NEAT1 慢病毒转染 B16F10 细胞。接下来,我们发现 PPB 通过 MTT 检测和建立小鼠皮下移植肿瘤模型,对黑色素瘤和 B16F10 细胞具有显著的增殖抑制作用;此外,通过划痕愈合试验、Transwell 试验和建立小鼠黑色素瘤肺转移模型,我们发现 PPB 显著抑制了 B16F10 细胞的体外侵袭和迁移,并抑制了黑色素瘤向肺、骨和肝的转移。最后,通过 Western blot(WB)和免疫组化评估 EMT 相关蛋白表达水平的变化,发现 PPB 显著下调了 MMP-9、N-钙粘蛋白等的表达水平,并上调了 E-钙粘蛋白。而过表达的 NEAT1 表现出促进黑色素瘤增殖、迁移和侵袭的能力,此外,还部分逆转了 PPB 对上述黑色素瘤增殖、迁移和侵袭的抑制作用。

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