Li Juan, Lian Yifan, Yan Changsheng, Cai Zeling, Ding Jie, Ma Zhonghua, Peng Peng, Wang Keming
The Second Clinical Medical College of Nanjing Medical University, Nanjing, Jiangsu, China.
Department of Oncology, Second Affiliated Hospital, Nanjing Medical University, Nanjing, Jiangsu, China.
Cell Prolif. 2017 Feb;50(1). doi: 10.1111/cpr.12312. Epub 2016 Oct 27.
Despite improvements in diagnosis and treatment, colorectal cancer (CRC) remains the third most common malignancy, and fourth-leading cause of cancer-related death worldwide, and has a particularly high incidence in Western countries. Recent studies have suggested that long non-coding RNAs (lncRNAs) compose a novel class of regulators of cancer biological processes, such as proliferation, apoptosis and metastasis. Here, we report that lncRNA FOXP4-AS1 acts as a functional oncogene in CRC pathogenesis. Moreover, we have attempted to investigate the effects of FOXP4-AS1 on tumour progression, both in vitro and in vivo.
In this study, bioinformatic analyses and qPCR were performed to investigate FOXP4-AS1 expression in CRC tissue samples and CRC cell lines. We inhibited FOXP4-AS1 expression via FOXP4-AS1-specific siRNA transfection. Cell proliferation was assessed using cell viability and colony formation assays, as well as by flow cytometry and ethynyl deoxyuridine (Edu) analyses. Apoptosis was assessed using flow cytometry. Animal tumour xenografts were generated, and immunohistochemistry (IHC) was performed to evaluate effects of FOXP4-AS1 on CRC tumour growth in vivo.
We found that FOXP4-AS1 was up-regulated in CRC tissues and cell lines and that its overexpression positively correlated with advanced pathological stages and larger tumour size. Additionally, we found that FOXP4-AS1 knockdown inhibited cell proliferation and induced apoptosis. Furthermore, FOXP4-AS1 knockdown induced marked increase in number of cells in G0/G1 phase and reduction in number of cells in S phase, in DLD-1, HT-29 and HCT116 cell lines. Consistent with these findings, FOXP4-AS1 silencing inhibited tumour growth in vivo.
These findings suggest that FOXP4-AS1 plays a crucial role in CRC progression and may be a new biomarker in patients with CRC.
尽管在诊断和治疗方面有所改善，但结直肠癌（CRC）仍是全球第三大常见恶性肿瘤，也是癌症相关死亡的第四大主要原因，在西方国家发病率尤其高。最近的研究表明，长链非编码RNA（lncRNAs）构成了一类新型的癌症生物学过程调节因子，如增殖、凋亡和转移。在此，我们报告lncRNA FOXP4-AS1在CRC发病机制中作为一种功能性癌基因发挥作用。此外，我们试图研究FOXP4-AS1在体外和体内对肿瘤进展的影响。
在本研究中，进行了生物信息学分析和qPCR以研究FOXP4-AS1在CRC组织样本和CRC细胞系中的表达。我们通过FOXP4-AS1特异性siRNA转染抑制FOXP4-AS1表达。使用细胞活力和集落形成试验以及流式细胞术和乙炔基脱氧尿苷（Edu）分析评估细胞增殖。使用流式细胞术评估凋亡。生成动物肿瘤异种移植模型，并进行免疫组织化学（IHC）以评估FOXP4-AS1对体内CRC肿瘤生长的影响。
我们发现FOXP4-AS1在CRC组织和细胞系中上调，其过表达与晚期病理阶段和更大的肿瘤大小呈正相关。此外，我们发现FOXP4-AS1敲低抑制细胞增殖并诱导凋亡。此外，在DLD-1、HT-29和HCT116细胞系中，FOXP4-AS1敲低导致G0/G1期细胞数量显著增加，S期细胞数量减少。与这些发现一致，FOXP4-AS1沉默在体内抑制肿瘤生长。
这些发现表明FOXP4-AS1在CRC进展中起关键作用，可能是CRC患者的一种新生物标志物。
