肿瘤防治研究2020年第47卷第11期 Cancer Res Prev Treat,2020,V ol.47,No.11·817·doi:10.3971/j.issn.1000-8578.2020.20.0069HNF4A 充当肿瘤抑制基因抑制结直肠肿瘤进展的实验
邵胜利，杨熹Experiment of HNF4A as a Tumor Suppressor Gene Inhibiting Progression of Colorectal Cancer
SHAO Shengli, YANG Xi
Department of Gastroenterology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, China词牌名有什么
CorrespondingAuthor:YANGXi,E-mail:****************
小名怎么取
Abstract: Objective To investigate the role of HNF4A in the progression of colorectal cancer. Methods We constructed colorectal cancer cell lines with HNF4A overexpression or knockdown. The expression level of HNF4A was detected by real-time quantitative PCR and Western blot; the colony forming ability of the cell lines were tested by soft agar and plate clone formation experiments, and the cell proliferation ability were checked by CCK-8 method; the expression le
vels of stem cell markers were detected by Western blot. The cell migration and invasion abilities were detected by Transwell migration and invasion experiments. The correlation between HNF4A and matrix metalloproteinas MMP2, MMP9 expression in colorectal cancer was analyzed by R2 gene platform and further verified by Western blot. Results The overexpression of HNF4A inhibited the proliferation, colony formation, migration and invasion of colorectal tumor cells. The knockdown of HNF4A promoted the proliferation, colony formation, migration and invasion of colorectal cancer cells. HNF4A inhibited the expression of colorectal cancer stem cells markers, such as CD133, CD44 and EpCAM, as well as the expression of MMP2 and MMP9. Moreover, the same conclusion was obtained in HNF4A-knockdown cell lines. Conclusion HNF4A inhibits the progression of colorectal cancer by inhibiting the stem-like cell characteristic and the expression of metalloproteinas.ekc
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母材Key words: HNF4A; Colorectal cancer; Proliferation; Stem-like cell characteristics; Migration; Invasion Funding: National Natural Science Foundation of China (No. 81802426)
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Competing interests: The authors declare that they have no competing interests.
摘 要：目的 探究HNF4A 在结直肠肿瘤进展中的作用。方法 构建HNF4A 过表达或敲除的结直肠癌
元游中国象棋细胞系，通过实时定量PCR 和Western blot 检测构建的细胞系HNF4A 的表达水平；通过软琼脂及平板克隆形成实验检测细胞系的克隆形成能力，CCK-8法检测细胞的增殖能力；Western blot 检测干细胞标志物的表达水平；最后通过Transwell 迁移与侵袭实验检测细胞的迁移侵袭能力，R2基因分析平台分析结直肠癌中HNF4A 与基质金属蛋白酶MMP2及MMP9表达的相关性并通过Western blot 验证。结果 过表达HNF4A 抑制了结直肠肿瘤细胞的增殖与克隆形成能力及迁移侵袭能力，敲低HNF4A 促进了结直肠癌细胞的增殖与克隆能力及迁移侵袭能力，Western blot 显示HNF4A 抑制了结直肠癌干细胞的标志物CD133、CD44及EpCAM 的表达和基质金属蛋白酶MMP2及MMP9的表达，在HNF4A 敲低的细胞系中得出了同样的结论。结论 HNF4A 通过抑制结直肠癌干细胞特征及金属蛋白酶的表达来抑制结直肠肿瘤的进展。
关键词：HNF4A ；结直肠癌；增殖；干细胞特征；迁移；侵袭
中图分类号：R735.3+5 开放科学(资源服务)标识码(OSID)：
收稿日期：2020-02-04；修回日期：2020-03-05基金项目：国家自然科学基金青年科学基金（81802426） 作者单位：430030 武汉，华中科技大学同济医学院附
属同济医院胃肠外科
通信作者：杨熹（1986-），男，博士，主治医师，主
梦见被人杀要从事结直肠肿瘤的研究，E-mail: ****************
作者简介：邵胜利（1994-）， 男，硕士在读，主要
从事结直肠肿瘤的研究·基础研究·0 引言全球癌症调查报告指出每年约有180万新发结直肠癌病例和86万结直肠肿瘤特异性死亡病例，分别占总发病例数和总死亡例数的10%和9%[1]。肿瘤进展是肿瘤特异性死亡的主要原因之一，多种机制参与了肿瘤进展，包括增长信号的自给自足、
对抑制生长信号不敏感、规避凋亡、无限的自我
