Han Chaofei, Tang Fengjie, Chen Jia, Xu Dan, Li Xiong, Xu Yangcheng, Wang Shaohua, Zhou Jianda
Department of Burn and Plastic Surgery, Third Xiangya Hospital of Central South University, Changsha, Hunan 410013, P.R. China.
Exp Ther Med. 2019 May;17(5):4294-4302. doi: 10.3892/etm.2019.7421. Epub 2019 Mar 20.
Long non-coding RNA urothelial carcinoma-associated 1 (UCA1) functions as an oncogene in different human cancers, including melanoma. However, the molecular mechanism of UCA1 underlying melanoma progression still remains largely unknown. In the present study, reverse transcription quantitative polymerase chain reaction and western blot analyses were used to examine the mRNA and protein expression levels, respectively. Cell Counting Kit-8 and wound healing assays were conducted to study cell proliferation and migration, respectively. A luciferase reporter assay was used to confirm the targeting relationship. It was demonstrated that UCA1 expression was increased in melanoma tissues and cell lines. In addition, UCA1 expression was higher in melanoma tissues at stage III-IV than in tissues at stage I-II. Inhibition of UCA1 expression markedly reduced melanoma cell proliferation and migration. Further investigation revealed that UCA1 functioned in melanoma cells through directly binding with microRNA (miR)-28-5p. The expression of miR-28-5p was significantly reduced in melanoma tissues and had an inverse correlation with UCA1 expression. In addition, miR-28-5p expression was higher in melanoma tissues at advanced stages than in stage I-II tissues. Furthermore, homeobox (HOX)B3 was identified as a target gene of miR-28-5p in melanoma cells, and HOXB3 overexpression reversed the suppressive effects of UCA1 downregulation on melanoma cell proliferation and migration. Finally, HOXB3 was upregulated in melanoma tissues compared with its expression in adjacent tissues, and HOXB3 expression was increased in melanoma tissues at advanced stages. Taken together, the regulatory network of the UCA1/miR-28-5p/HOXB3 axis in melanoma was demonstrated for the first time in the present study, expanding the understanding of the molecular mechanism underlying melanoma progression. Future studies may further confirm the function of this signaling pathway .
长链非编码RNA尿路上皮癌相关1(UCA1)在包括黑色素瘤在内的不同人类癌症中发挥致癌基因的作用。然而,UCA1在黑色素瘤进展中的分子机制仍 largely 未知。在本研究中,分别使用逆转录定量聚合酶链反应和蛋白质印迹分析来检测mRNA和蛋白质表达水平。分别进行细胞计数试剂盒-8和伤口愈合试验以研究细胞增殖和迁移。使用荧光素酶报告基因试验来确认靶向关系。结果表明,UCA1在黑色素瘤组织和细胞系中的表达增加。此外,III-IV期黑色素瘤组织中UCA1的表达高于I-II期组织。抑制UCA1表达显著降低黑色素瘤细胞的增殖和迁移。进一步研究表明,UCA1通过直接与微小RNA(miR)-28-5p结合在黑色素瘤细胞中发挥作用。miR-28-5p在黑色素瘤组织中的表达显著降低,且与UCA1表达呈负相关。此外,晚期黑色素瘤组织中miR-28-5p的表达高于I-II期组织。此外,同源框(HOX)B3被鉴定为黑色素瘤细胞中miR-28-5p的靶基因,HOXB3过表达逆转了UCA1下调对黑色素瘤细胞增殖和迁移的抑制作用。最后,与相邻组织相比,HOXB3在黑色素瘤组织中上调,且晚期黑色素瘤组织中HOXB3的表达增加。综上所述,本研究首次证明了黑色素瘤中UCA1/miR-28-5p/HOXB3轴的调控网络,扩展了对黑色素瘤进展分子机制的理解。未来的研究可能会进一步证实该信号通路的功能。