Department of Dermatology, Zhejiang Provincial People's Hospital, People's Hospital of Hangzhou Medical College, Hangzhou, China.
Eur Rev Med Pharmacol Sci. 2020 Sep;24(17):8940-8946. doi: 10.26355/eurrev_202009_22835.
The aim of this study was to elucidate the role of FOXC2-AS1 in promoting the proliferative ability and inhibiting apoptosis of melanoma by silencing p15, thereafter regulating the progression of melanoma.
FOXC2-AS1 levels in melanoma patients with or without metastasis and those with the tumor in different stages were detected by quantitative real-time polymerase chain reaction (qRT-PCR). Regulatory effects of FOXC2-AS1 on viability and apoptosis in melanoma cells were assessed, and subcellular distribution of FOXC2-AS1 was analyzed. Subsequently, the interactions of FOXC2-AS1 with EZH2 and SUZ12 were explored by RNA-Binding Protein Immunoprecipitation (RNA-RIP) assay. Through chromatin immunoprecipitation (ChIP) assay, the role of FOXC2-AS1 to regulate p15 transcription by recruiting EZH2 was verified. At last, regulatory effects of FOXC2-AS1/p15 axis on viability and apoptosis in melanoma cells were investigated.
It was found that FOXC2-AS1 was upregulated in melanoma tissues, especially those with metastasis or stage II-IV. Melanoma patients expressing high level of FOXC2-AS1 showed worse survival than those with low level. Knockdown of FOXC2-AS1 inhibited viability, and stimulated apoptosis in A375 and sk-mel-110 cells. Besides, P15 level was upregulated in melanoma cells transfected with si-FOXC2-AS1, and FOXC2-AS1 was mainly distributed in cytoplasm. RNA-RIP assay confirmed that FOXC2-AS1 was mainly enriched in anti-EZH2 and aniti-SUZ12. Knockdown of EZH2 could markedly upregulate protein level of p15 in melanoma cells. Furthermore, it was verified that FOXC2-AS1 inhibited p15 transcription via recruiting EZH2, and the knockdown of p15 could partially reverse the regulatory effects of FOXC2-AS1 on viability and apoptosis in melanoma.
FOXC2-AS1 stimulates proliferative ability in melanoma via silencing p15.
本研究旨在通过沉默 p15 来阐明 FOXC2-AS1 在促进黑色素瘤增殖能力和抑制细胞凋亡中的作用，进而调节黑色素瘤的进展。
通过实时定量聚合酶链反应（qRT-PCR）检测有或无转移以及处于不同阶段的黑色素瘤患者的 FOXC2-AS1 水平。评估 FOXC2-AS1 对黑色素瘤细胞活力和凋亡的调节作用，并分析 FOXC2-AS1 的亚细胞分布。随后，通过 RNA 结合蛋白免疫沉淀（RNA-RIP）实验探讨 FOXC2-AS1 与 EZH2 和 SUZ12 的相互作用。通过染色质免疫沉淀（ChIP）实验验证 FOXC2-AS1 通过募集 EZH2 来调节 p15 转录的作用。最后，研究了 FOXC2-AS1/p15 轴对黑色素瘤细胞活力和凋亡的调节作用。
发现 FOXC2-AS1 在黑色素瘤组织中上调，特别是在有转移或 II-IV 期的黑色素瘤组织中。表达高水平 FOXC2-AS1 的黑色素瘤患者的生存状况比表达低水平 FOXC2-AS1 的患者差。敲低 FOXC2-AS1 可抑制 A375 和 sk-mel-110 细胞的活力，并刺激细胞凋亡。此外，转染 si-FOXC2-AS1 的黑色素瘤细胞中 P15 水平上调，FOXC2-AS1 主要分布在细胞质中。RNA-RIP 实验证实 FOXC2-AS1 主要富集在抗 EZH2 和抗 SUZ12 中。敲低 EZH2 可显著上调黑色素瘤细胞中 p15 蛋白水平。此外，证实 FOXC2-AS1 通过募集 EZH2 抑制 p15 转录，敲低 p15 可部分逆转 FOXC2-AS1 对黑色素瘤细胞活力和凋亡的调节作用。
FOXC2-AS1 通过沉默 p15 刺激黑色素瘤的增殖能力。
