天津医药

天津医药 ›› 2021, Vol. 49 ›› Issue (4): 354-358.doi: 10.11958/20202872

• 细胞与分子生物学 • 上一篇    下一篇

lncRNA FOXCUT调控Notch通路抑制结肠癌细胞增殖、侵袭的实验研究#br#

佘明豪1，刘寒松1，杨文辉1，余洋1，张磊2   

  1. 1郑州大学附属郑州中心医院普通外科（邮编450000），2消化内科
  • 收稿日期:2020-10-21 修回日期:2020-12-22 出版日期:2021-04-15 发布日期:2021-04-16
  • 作者简介:佘明豪（1981），男，本科，主治医师，主要从事肝胆胰、胃肠结直肠、腹部外科诊治研究。E-mail：mimiho81@163.com
  • 基金资助:
    河南省医学科技攻关计划联合共建项目（LHGJ20191060）

Experimental study of lncRNA FOXCUT regulating Notch pathway to inhibit proliferation and invasion of colon cancer cells

SHE Ming-hao1, LIU Han-song1, YANG Wen-hui1, YU Yang1, ZHANG Lei2   

  1. 1 Department of General Surgery, 2 Department of Gastroenterology, Zhengzhou Central Hospital Affiliated to Zhengzhou University, Zhengzhou 450000, China
  • Received:2020-10-21 Revised:2020-12-22 Published:2021-04-15 Online:2021-04-16

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摘要： 目的　研究长链非编码RNA（lncRNA）FOXC1启动子临近转录体（FOXCUT）对结肠癌细胞（HCT116）增殖、侵袭的影响及作用机制。方法　向HCT116细胞转染FOXCUT siRNA（FOXCUT siRNA组），以转染阴性siRNA为阴性对照组（FOXCUT siRNA-NC组），未转染组为正常对照组（NC组），通过qPCR法确定FOXCUT干扰效果，并检测结肠癌及癌旁组织、人结肠癌细胞株（SW480、SW620、HCT116、HT29）和人正常结肠上皮细胞（NCM460）中FOXCUT mRNA表达水平；通过CCK-8、集落形成实验和Transwell实验检测沉默FOXCUT对HCT116细胞增殖、集落形成能力及侵袭能力的影响；qPCR检测沉默FOXCUT后HCT116细胞中Notch1、Hes1 mRNA表达变化；蛋白免疫印迹（Western blot）实验检测HCT116细胞中Notch1、Hes1蛋白表达情况。结果　与癌旁组织比较，结肠癌组织中FOXCUT mRNA表达水平显著增高（P＜0.01）；与NCM460细胞比较，SW480、SW620、HCT116、HT29结肠癌细胞株中FOXCUT mRNA表达水平均显著增高（P＜0.05），其中以HCT116细胞中表达水平最高；与FOXCUT siRNA-NC组比较，FOXCUT siRNA组HCT116细胞中FOXCUT mRNA表达水平显著降低（P＜0.01）；NC组、FOXCUT siRNA-NC组HCT116细胞中FOXCUT mRNA表达水平差异无统计学意义（P＞0.05）；沉默FOXCUT可显著抑制细胞增殖、集落形成能力及侵袭（P＜0.05），并抑制Notch1、Hes1 mRNA及蛋白表达（P＜0.01）。结论　沉默FOXCUT可抑制结肠癌HCT116细胞增殖及侵袭，可能与抑制Notch通路有关。

关键词: RNA, 长链非编码, 结肠肿瘤, , HCT116细胞, 细胞增殖, 肿瘤侵润, 受体, Notch1, 转录因子HES-1

Abstract: Objective　To study the effect of long non-coding RNA (lncRNA) FOXC1 promoter upstream transcript (FOXCUT) on the proliferation and invasion of colon cancer cell (HCT116) and its mechanism. Methods　 HCT116 cells transfected with FOXCUT siRNA were used as FOXCUT siRNA group. The transfection-negative interfering RNA was used as the negative control group (FOXCUT siRNA-NC group), and the untransfected group was used as the normal control group (NC group). The qPCR method was used to determine the FOXCUT interference effect and to detect the mRNA expression levels of FOXCUT in colon cancer and adjacent tissues, human colon cancer (SW480, SW620, HCT116, HT29) and human normal colon epithelial cells (NCM460). The effects of silencing FOXCUT on the proliferation and the invasion of HCT116 cells were detected by Cell Counting Kit-8 (CCK-8), colony forming assay and Transwell assay. qPCR was used to detect the mRNA expressions of Notch1 and Hes1 in HCT116 cells after FOXCUT silencing, and Western blot assay was used to detect the protein expressions of Notch1 and Hes1 in HCT116 cells. Results　Compared with adjacent tissues, the expression level of FOXCUT mRNA increased significantly in colon cancer tissues (P＜0.01). Compared with NCM460 cells, the FOXCUT mRNA expression levels increased significantly in colon cancer cell lines SW480, SW620, HCT116, and HT29 (P＜0.05), and the expression level was the highest in HCT116 cells. Compared with the FOXCUT siRNA-NC group, the FOXCUT mRNA expression levels were significantly reduced in HCT116 cells of the FOXCUT siRNA group (P＜0.01). There was no significant difference in the FOXCUT mRNA expression level in the HCT116 cells between the NC group and FOXCUT siRNA-NC group (P＞0.05). The silencing FOXCUT could significantly inhibit the proliferation and invasion of HCT116 cells (P＜0.05), and inhibit the mRNA and protein expressions of Notch1 and Hes1 in HCT116 cells (P＜0.01). Conclusion　Silencing FOXCUT can inhibit the proliferation and invasion of HCT116 cells, which may be related to the inhibition of Notch pathway.

Key words: RNA, long noncoding, colonic neoplasms, HCT116 cells, cell proliferation, neoplasm invasiveness, receptor, Notch1, transcription factor HES-1

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