lncRNA CASC2对结直肠癌细胞功能及血管生成的影响

doi:10.11904/j.issn.1002-3070.2022.02.003

实用肿瘤学杂志 ›› 2022, Vol. 36 ›› Issue (2): 111-118.doi: 10.11904/j.issn.1002-3070.2022.02.003

lncRNA CASC2对结直肠癌细胞功能及血管生成的影响

陈函¹, 罗晓勇^2,3, 李朝萍^2,3, 郭彩茹⁴, 鞠文翠^2,3, 翟秧秧^2,3, 师鑫鹏^2,3

1.郑州大学第二附属医院(郑州 450014);
2.洛阳市肿瘤临床诊疗研究中心;
3.郑州大学附属洛阳中心医院放疗科;
4.河南省肿瘤免疫与再生医学国际联合实验室

收稿日期:2021-09-30 修回日期:2021-12-30 出版日期:2022-04-28 发布日期:2022-04-28
通讯作者: 罗晓勇,E-mail:xyluo68@163.com
作者简介:陈函,女,(1996-),硕士研究生,从事消化道肿瘤发病机制的研究。
基金资助:
2020年河南省医学科技攻关计划项目(编号:202102310421);2020年度洛阳市医疗卫生领域指导性科技计划项目(编号:2020002A)

The effect of lncRNA CASC2 on the function and angiogenesis of colorectal cancer cells

CHEN Han¹, LUO Xiaoyong^2,3, LI Chaoping^2,3, GUO Cairu⁴, JU Wencui^2,3, ZHAI Yangyang^2,3, SHI Xinpeng^2,3

1. The Second Affiliated Hospital of Zhengzhou University,Zhengzhou 450014,China;
2. Luoyang Cancer Clinical Diagnosis and Treatment Center;
3. Department of Radiation Oncology,Luoyang Central Hospital Affiliated to Zhengzhou University;
4. Henan International Joint Laboratory of Tumor Immunology and Regenerative Medicine

Received:2021-09-30 Revised:2021-12-30 Online:2022-04-28 Published:2022-04-28

摘要/Abstract

摘要： 目的研究lncRNA CASC2过表达对结直肠癌细胞增殖、迁移、血管生成及Wnt/FZD/β-catenin信号通路的影响。方法 qRT-PCR检测lncRNA CASC2在正常结肠上皮细胞(NCM460)及结直肠癌细胞株(DLD-1、HT29、SW620、HCT116、RKO、LOVO、SW480)中的表达情况;HCT116和SW480细胞建立稳定转染的细胞株,分为对照组和lncRNA CASC2过表达组;qRT-PCR验证稳转株中lncRNA CASC2水平;CCK-8实验、Transwell实验、划痕实验分别检测细胞增殖水平、迁移能力;收集对照组和lncRNA CASC2过表达组细胞上清并提取外泌体,将对照组和lncRNA CASC2过表达组细胞的外泌体分别与HUVEC细胞共孵育,3D Matrigel法检测其对血管生成的影响;Western blot测定细胞中Wnt16、FZD7、β-catenin蛋白水平。结果 lncRNA CASC2在7株结直肠癌细胞株中的表达明显低于正常结肠上皮细胞NCM460细胞。与对照组相比,lncRNA CASC2过表达后明显抑制了HCT116和SW480细胞的增殖、迁移能力(P＜0.01)。3D Matrigel法检测显示,来自SW480-CASC2组、HCT116-CASC2组细胞的外泌体显著抑制了HUVEC中管状细胞的形成(P＜0.01)。Western blot结果显示,lncRNA CASC2过表达后Wnt16、FZD7、β-catenin蛋白水平降低(P＜0.01)。结论 lncRNA CASC2在结直肠癌细胞中表达下调,lncRNA CASC2过表达可以抑制Wnt/FZD/β-catenin信号通路进而抑制结直肠癌细胞的增殖、迁移,并减少血管生成。

关键词: 长链非编码RNA CASC2, 结直肠癌, 外泌体, Wnt信号通路, 血管生成

Abstract: Objective The of this study was to investigate the effect of lncRNA CASC2 overexpression on the proliferation,migration,angiogenesis and Wnt/FZD/β-catenin signaling pathway of colorectal cancer cells.Methods qRT-PCR was used to detect the expression of lncRNA CASC2 in normal colon epithelial cells(NCM460)and colorectal cancer cell lines(DLD-1,HT29,SW620,HCT116,RKO,LOVO,and SW480).HCT116 cells and SW480 cells were established stable transfection.The transfected cell lines were divided into the control group and lncRNA CASC2 overexpression group.CCK-8,Transwell,and scratch assays were used to detect the cell proliferation and migration ability.The cell supernatants collected from the control group and the lncRNA CASC2 overexpression group were used to extract exosomes,which were incubated with HUVEC cells.Their effects on angiogenesis were detected by 3D Matrigel method.The protein expression of Wnt16,FZD7 and β-catenin were determined by Western blot.Results The expression of lncRNA CASC2 in 7 colorectal cancer cell lines was significantly lower than that in normal colon epithelial NCM460 cells.Compared with the control group,the overexpression of lncRNA CASC2 significantly inhibited the proliferation,migration of HCT116 cells and SW480 cells(P＜0.01).3D Matrigel assay showed that exosomes from cells in SW480-CASC2 and HCT116-CASC2 groups significantly inhibited the formation of tubular cells in HUVEC(P＜0.01).The results of Western blot showed that the levels of Wnt16,FZD7 and β-catenin protein decreased after lncRNA CASC2 overexpression(P＜0.01).Conclusion The expression of lncRNA CASC2 is down-regulated in colorectal cancer cells.Overexpression of lncRNA CASC2 can inhibit the proliferation and migration of colorectal cancer cells by inhibiting the Wnt/FZD/β-catenin signaling pathway,and reducing angiogenesis.

Key words: Long non-coding RNA CASC2, Colorectal cancer, Exosomes, Wnt signaling pathway, Angiogenesis

中图分类号:

R735.3

陈函, 罗晓勇, 李朝萍, 郭彩茹, 鞠文翠, 翟秧秧, 师鑫鹏. lncRNA CASC2对结直肠癌细胞功能及血管生成的影响[J]. 实用肿瘤学杂志, 2022, 36(2): 111-118.

CHEN Han, LUO Xiaoyong, LI Chaoping, GUO Cairu, JU Wencui, ZHAI Yangyang, SHI Xinpeng. The effect of lncRNA CASC2 on the function and angiogenesis of colorectal cancer cells[J]. Journal of Practical Oncology, 2022, 36(2): 111-118.

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lncRNA CASC2对结直肠癌细胞功能及血管生成的影响

The effect of lncRNA CASC2 on the function and angiogenesis of colorectal cancer cells

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