期刊导航

论文摘要

晚期糖基化终末产物受体在胰腺癌细胞增殖及 成瘤过程中的作用

作者:陈巍巍, 郭强, 张肇达等

Author:CHEN Wei-wei, GUO Qiang, ZHANG Zhao-da. et al

收稿日期:          年卷(期)页码:2017,48(1):46-51

期刊名称:四川大学学报(医学版)

Journal Name:JOURNAL OF SICHUAN UNIVERSITY (MEDICAL SCIENCE EDITION)

关键字:晚期糖基化终末产物受体 胰腺癌 RNA干扰

Key words:RAGE Pancreatic cancer RNA interference

基金项目:

中文摘要

目的 探讨晚期糖基化终末产物受体(receptor for advanced glycation end products, RAGE)对胰腺癌PANC-1细胞增殖及转染后接种裸鼠在成瘤过程中的作用。方法 构建RAGE 短发夹RNA (shRNA),即shRNA RAGE -1、-2、-3质粒,转染胰腺癌PANC-1细胞后,采用八肽胆囊收缩素(CCK-8)法、逆转录聚合酶链反应(RT-PCR)和Western blot检测shRNA RAGE 对细胞增殖、RAGE表达的影响,筛选干扰效果最好的shRNA RAGE 质粒;将转染了shRNA RAGE 质粒的PANC-1细胞接种于裸鼠皮下,观察成瘤时间,并测量计算肿瘤体积;采用RT-PCR和Western blot检测shRNA RAGE 对RAGE、基质金属蛋白酶 (matrix metalloproteinase, MMP)2和9(MMP-2和MMP-9)、核因子-κB(nuclear factor kappa-light-chain-enhancer of activated B cells, NF-κB)、血管内皮生长因子(vascular endothelial growth factor, VEGF )的mRNA及蛋白表达的影响,并采用免疫组化方法对肿瘤进行微血管计数,计算微血管密度。结果 转染shRNA RAGE 24 h后CCK-8检测细胞的吸光度(A)值低于对照组( PRAGE -2质粒转染后下调RAGE表达效果最好。转染shRNA RAGE -2质粒的PANC-1细胞接种于裸鼠后,裸鼠成瘤时间、肿瘤体积低于shRNA RAGE -1,-3,裸鼠肿瘤组织RAGE 、MMP-2、 NF-κB、 MMP-9、 VEGF mRNA及蛋白表达低于shRNA RAGE -1,-3( PRAGE -1,-3( P

英文摘要

Objective To investigate the effect of receptor for advanced glycation end products (RAGE) on cell proliferation and tumor growth in nude mice with pancreatic cancer. Methods PANC-1 cells were transfected with shRNA RAGE -1,-2,-3 to down-regulate the expression of RAGE. Cholecystokinin octopeptide-8 (CCK-8), real-time PCR and Western blot were performed to test the impact of shRNA RAGE on the expressions of mRNAs and proteins of RAGE, matrix metalloproteinase-2 (MMP-2), MMP-9, nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB), and vascular endothelial growth factor (VEGF). Tumor growth and microvessel density in the nude mice implanted with shRNA RAGE transfected PANC-1 cells were observed using immunohistochemistry. Results The shRNA RAGE -1,-2,-3 transfected cells had lower absorbance values than the controls 24 h after transfection, and the absorbance value reached the lowest at 48 h. The specific shRNA sequences significantly inhibited the expressions of mRNA and protein of RAGE. The mice implanted with shRNA RAGE -2 had lower tumor volume and microvessel density than shRNA RAGE -1,-3. The expressions of mRNAs and proteins of RAGE, MMP-2, NF-κB, MMP-9 and VEGF were lower in the cells transfected with shRNA RAGE -2 compared with shRNA RAGE -1,-3. Conclusion RAGE is involved in the progression of pancreatic cancerin vitro and in vivo . The RAGE expression could influence the process of tumor angiogenesis.

关闭

Copyright © 2020四川大学期刊社 版权所有.

地址:成都市一环路南一段24号

邮编:610065