HOXA6-HOXA5 基因座上的一个新型长非编码 RNA 促进结肠癌细胞生长。
A novel long non-coding RNA from the HOXA6-HOXA5 locus facilitates colon cancer cell growth.
机构信息
Department of Pathophysiology, Institute of Biomedical Sciences, Tokushima University Graduate School, 3-18-15 Kuramoto-cho, Tokushima, 770-8503, Japan.
Department of Human Genetics, Institute of Biomedical Sciences, Tokushima University Graduate School, 3-18-15 Kuramoto-cho, Tokushima, 770-8503, Japan.
出版信息
BMC Cancer. 2019 Jun 3;19(1):532. doi: 10.1186/s12885-019-5715-0.
BACKGROUND
Homeobox A5 (HOXA5), a member of the HOX family, plays an important role in tumor development and morphogenesis, although opposite effects on tumorigenesis have been observed, depending on the tissue type. In this study, we aimed to investigate the role of a novel transcript from the HOXA6-HOXA5 locus in colon cancer tumorigenesis.
METHODS
Human colon cancer cell lines were analyzed using next generation sequencing-based targeted mRNA capture. The effects of overexpression and silencing of HOXA5 transcripts were evaluated in vitro and using a xenograft nude mouse model.
RESULTS
We identified three novel transcripts (HOXA5 short, long 1, and long 2) transcribed from the HOXA6-HOXA5 locus in HCT116 colon cancer cells using next generation sequencing-based targeted mRNA capture. Knockdown of HOXA5 long 1 and long 2 transcripts did not affect cell growth, while selective silencing of HOXA5 short RNA inhibited cell growth independent of HOXA5 expression. Stable overexpression of HOXA5 short RNA promoted proliferation and migration of colon cancer cell lines HCT116, DLD1, and HT-29 and accelerated tumor growth in the xenograft mouse model. In vitro translation assays suggested HOXA5 short RNA was a functional long non-coding RNA (lncRNA). Consistent with these observations, expression of HOXA5 short RNA was upregulated in advanced colon cancer tissues. Ingenuity Pathway Analysis of differentially expressed genes between HOXA5 short RNA overexpressed and silenced HCT116 cells revealed that HOXA5 short RNA preferentially modified expression of epidermal growth factor (EGF) signal-related genes. Western blot analysis demonstrated that stable overexpression of HOXA5 short RNA increased EGF receptor levels and facilitated its phosphorylation in both HCT116 cells and xenograft tumors.
CONCLUSIONS
Our results suggested that HOXA5 short RNA, a novel lncRNA, may play a crucial role in colon tumor growth through activation of EGF signaling.
背景
同源盒 A5（HOXA5）是同源盒家族的成员，在肿瘤发生和形态发生中发挥重要作用，尽管根据组织类型观察到对肿瘤发生的相反影响。在这项研究中，我们旨在研究 HOXA6-HOXA5 基因座的新型转录物在结肠癌肿瘤发生中的作用。
方法
使用基于下一代测序的靶向 mRNA 捕获分析人结肠癌细胞系。在体外和异种移植裸鼠模型中评估 HOXA5 转录物过表达和沉默的影响。
结果
我们使用基于下一代测序的靶向 mRNA 捕获技术在 HCT116 结肠癌细胞中鉴定出三个新型转录物（HOXA5 短、长 1 和长 2）从 HOXA6-HOXA5 基因座转录。HOXA5 长 1 和长 2 转录物的敲低不影响细胞生长，而选择性沉默 HOXA5 短 RNA 抑制细胞生长而不依赖 HOXA5 表达。HOXA5 短 RNA 的稳定过表达促进结肠癌细胞系 HCT116、DLD1 和 HT-29 的增殖和迁移，并加速异种移植小鼠模型中的肿瘤生长。体外翻译实验表明 HOXA5 短 RNA 是一种功能性长非编码 RNA（lncRNA）。与这些观察结果一致，HOXA5 短 RNA 在晚期结肠癌组织中表达上调。HOXA5 短 RNA 过表达和沉默的 HCT116 细胞中差异表达基因的 IPA 分析表明，HOXA5 短 RNA 优先修饰表皮生长因子（EGF）信号相关基因的表达。Western blot 分析表明，HOXA5 短 RNA 的稳定过表达增加了 HCT116 细胞和异种移植肿瘤中 EGF 受体的水平，并促进了其磷酸化。
结论
我们的结果表明，HOXA5 短 RNA，一种新型 lncRNA，可能通过激活 EGF 信号在结肠肿瘤生长中发挥关键作用。
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