Lu Canrong, Xie Tianyu, Guo Xin, Wu Di, Li Shuo, Li Xiongguang, Lu Yixun, Wang Xinxin
Department of General Surgery, Chinese People's Liberation Army General Hospital, Beijing 100853, People's Republic of China.
Cancer Manag Res. 2020 Jun 9;12:4347-4356. doi: 10.2147/CMAR.S250670. eCollection 2020.
Long non-coding RNA (lncRNA) DSCAM-AS1 was reported to be aberrantly expressed and play pivotal roles in various human cancers. The aim of the present study was to investigate the expression and roles of DSCAM-AS1 in colon cancer (CC).
Quantitative real-time PCR (qRT-PCR) was used to detect the expression of DSCAM-AS1, miR-204 and the mRNA level of SOX4. Cell proliferation and cell cycle were analyzed by MTT assay and flow cytometry, respectively. Transwell assay was used for migration capacity detection. Luciferase activity assay was conducted to verify the direct binding of DSCAM-AS1 and miR-204 or miR-204 and SOX4. The protein expression of SOX4 was determined by Western blot. Kaplan-Meier curves were calculated and the Log rank test was performed for the survival data analysis.
DSCAM-AS1 was significantly upregulated in CC and high expression of DSCAM-AS1 was associated with poor prognosis in patients with colon cancer. Knockdown of DSCAM-AS1 significantly suppressed CC cells proliferation and migration. In addition, DSCAM-AS acted as a molecular sponge for miR-204 and SOX4 was identified as a direct target of miR-204 in CC. Moreover, the rescue assay revealed that miR-204 inhibition partly abolished the effects of DSCAM-AS1 knockdown on CC cells proliferation, migration and SOX4 expression.
The present study demonstrated that DSCAM-AS1 acted as an oncogenic lncRNA in CC progression by regulating miR-204/SOX4 axis and DSCAM-AS1 may serve as a novel therapeutic target in the treatment of colon cancer.
据报道,长链非编码RNA(lncRNA)DSCAM-AS1在多种人类癌症中异常表达并发挥关键作用。本研究旨在探讨DSCAM-AS1在结肠癌(CC)中的表达及作用。
采用定量实时PCR(qRT-PCR)检测DSCAM-AS1、miR-204的表达及SOX4的mRNA水平。分别通过MTT法和流式细胞术分析细胞增殖和细胞周期。采用Transwell实验检测迁移能力。进行荧光素酶活性实验以验证DSCAM-AS1与miR-204或miR-204与SOX4之间的直接结合。通过蛋白质免疫印迹法测定SOX4的蛋白表达。计算Kaplan-Meier曲线,并对生存数据分析进行对数秩检验。
DSCAM-AS1在CC中显著上调,且DSCAM-AS1的高表达与结肠癌患者的不良预后相关。敲低DSCAM-AS1可显著抑制CC细胞的增殖和迁移。此外,DSCAM-AS充当miR-204的分子海绵,且SOX4被鉴定为CC中miR-204的直接靶点。此外,挽救实验表明,抑制miR-204可部分消除DSCAM-AS1敲低对CC细胞增殖、迁移及SOX4表达的影响。
本研究表明,DSCAM-AS1通过调节miR-204/SOX4轴在CC进展中作为致癌lncRNA发挥作用,且DSCAM-AS1可能成为结肠癌治疗的新靶点。