摘要/Abstract
摘要： 目的·研究蛋白质精氨酸甲基转移酶4（protein arginine methyltransferase 4，PRMT4）对胃癌发生和发展的影响及潜在的调控分子机制。方法·使用免疫组织化学技术检测PRMT4在32对胃癌和癌旁组织中的表达；利用公开数据库分析PRMT4在胃癌中的表达及其对胃癌患者生存期的影响；使用质粒介导的干扰技术下调胃癌细胞株MGC-803和MKN-45中PRMT4的表达，使用质粒过表达技术上调胃癌细胞株SGC-7901中PRMT4的表达；利用细胞生物学手段，如MTT、平板克隆形成、流式细胞术以及Transwell实验研究PRMT4对胃癌细胞增殖、凋亡以及侵袭与转移的影响；采用荧光素酶报告实验检测干扰PRMT4后细胞内源性转录因子TCF（T cell factor）转录活性的变化；使用实时定量荧光PCR检测干扰PRMT4后Wnt/β-catenin信号通路下游靶基因水平变化；使用蛋白质印迹法检测干扰及过表达PRMT4后Wnt/β-catenin信号通路的蛋白水平变化。结果·PRMT4蛋白在胃癌组织中表达上调，高表达PRMT4的胃癌患者生存期显著缩短。干扰PRMT4抑制胃癌细胞增殖、侵袭和转移，促进细胞凋亡，且抑制胃癌Wnt/β-catenin信号通路；过表达PRMT4促进胃癌细胞增殖、侵袭和转移，抑制细胞凋亡，且促进胃癌Wnt/β-catenin信号通路。结论·PRMT4在胃癌中过表达并且可能通过Wnt/β-catenin信号通路促进胃癌细胞生长、侵袭和转移。
关键词: 蛋白质精氨酸甲基转移酶4, 胃癌, 增殖, Wnt/β-catenin信号通路
Abstract:
Objective · To investigate the effect of protein arginine methyltransferase 4 (PRMT4) on the development and progression of gastric carcinoma and the underlying molecular mechanisms. Methods · The expression levels of PRMT4 in 32 specimens of gastric cancer and adjacent tissues were measured by immunohistochemistry technology. Public databases were used to analyze the expression of PRMT4 in gastric cancer and its effect on the survival of patients with gastric cancer. PRMT4 expressions of gastric cancer MGC-803 and MKN-45 cells were down-regulated by short hairpin RNA-mediated knockdown, and PRMT4 expression of gastric cancer SGC-7901 cells was up-regulated by using plasmid-mediated overexpression. Cell biology methods, such as MTT assay, colony formation assay, flow cytometry, and Transwell assay were used to study the effect of PRMT4 on proliferation, apoptosis, migration and invasion of gastric cancer cells. Luciferase reporter assay was used to detect transcription activity of endogenous transcription factor TCF (T cell factor) after PRMT4 silencing. The mRNA level changes of Wnt/β-catenin pathway downstream target genes after PRMT4 silencing were detected by real-time PCR. The protein level changes of Wnt/β-catenin pathway-related proteins after PRMT4 silencing and overexpression were detected by Western blotting. Results · PRMT4 was highly expressed in gastric cancer tissues. Patients with higher expression of PRMT4 had shorter survival than those with lower expression of PRMT4. The proliferation, colony formation, migration and invasion ability of gastric cancer cells were inhibited; the cell apoptosis was induced by silencing of PRMT4. The Wnt/β-catenin signaling pathway was also inhibited by PRMT4 knockdown. The proliferation, colony formation, migration and invasion ability of gastric cancer cells were induced; the cell apoptosis was inhibited by overexpressed PRMT4. The Wnt/β-catenin signaling pathway was also activated by PRMT4 overexpression. Conclusion · PRMT4 is highly expressed in gastric cancer. It can promote the growth, migration and invasion of gastric cancer cells through Wnt/β-catenin signaling pathway.
Key words: protein arginine methyltransferase 4 (PRMT4), gastric cancer, proliferation, Wnt/β-catenin signaling pathway
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