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

发布时间：2018-03-27 17:18

本文选题：晚期糖基化终末产物受体　切入点：胰腺癌　出处：《四川大学学报(医学版)》2017年01期

【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shRNARAGE-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shRNARAGE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shRNARAGE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shRNARAGE24h后CCK-8检测细胞的吸光度(A)值低于对照组(P0.05),且在转染48h时最明显。RT-PCR和Western blot检测显示,shRNA RAGE-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shRNARAGE-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(P0.05),其微血管密度也低于shRNARAGE-1,-3(P0.001)。结论 RAGE参与了胰腺癌的发生发展过程。RAGE可影响胰腺癌肿瘤生长及微血管形成。
[Abstract]:Objective to investigate the effect of advanced glycosylation end product receptor for advanced glycation end products on the proliferation of pancreatic cancer PANC-1 cells and the effect of transfection in nude mice. Methods the short hairpin RNA-shRNA-shRNA-shRNA-shRNA-3 plasmid was constructed and transfected into PANC-1 cells of pancreatic cancer. Using CCK-8 method, reverse transcriptase polymerase chain reaction (RT-PCR) and Western blot were used to detect the effect of shRNA RAGE on cell proliferation and rage expression. The shRNA RAGE plasmid with the best interfering effect was screened, and the PANC-1 cells transfected with shRNARAGE plasmid were inoculated subcutaneously into nude mice. RT-PCR and Western blot were used to detect the effect of shRNARAGE on mRNA and protein expression of RAGE, matrix metalloproteinase 2, 9(MMP-2 and MMP-9, nuclear factor- 魏 1# factor kappa-light-chain-enhancer of activated B cells of NF- 魏 B, vascular endothelial growth factor (VEGF) and vascular endothelial growth factor (VEGF) endothelial growth factor (VEGF). The microvessels of the tumor were counted by immunohistochemical method. Results after transfection of shRNARAGE24h, the absorbance of CCK-8 was lower than that of control group (P0.05A). The results of RT-PCR and Western blot analysis showed that the expression of RAGE was the best after transfection of RAGE-2 plasmid. ShRNA RAGE-2 was transfected with shRNA RAGE-2. The granulocyte PANC-1 cells were inoculated in nude mice. The tumorigenesis time and tumor volume of nude mice were lower than that of shRNARAGE-1 / 3, and the expression of VEGF mRNA and protein in tumor tissue of nude mice was lower than that of shRNA RAGE-1 / 3P0. 05, and the microvessel density was lower than that of shRNARAGE-1- 3 / P0. 001.Conclusion RAGE may play an important role in the development of pancreatic carcinoma. Rage can affect the growth and microangiogenesis of pancreatic carcinoma.
【作者单位】：四川大学华西医院胰腺外科;四川大学华西医院血管外科;
【基金】：四川省科技厅科技支撑计划项目(No.2013FZ0020)资助
【分类号】：R735.9

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