DH-332通过TRIM40/Raf-1/ERK1/2轴在胃癌中的作用和机制研究  

Study on the role and mechanism of DH-332 in gastric cancer through TRIM40/Raf-1/ERK1/2 axis

作　　者：李南 何文婷[1] 帕提玛·阿布力米提 轩艳红 赵俊涛 张洪亮[1] Li Nan;He Wenting;Patima·Abulimiti;Xuan Yanhong;Zhao Juntao;Zhang Hongliang(Oncology Department,The Fourth Affiliated Hospital of Xinjiang Medical University,Urumchi 830001,China)

机构地区：[1]新疆医科大学第四附属医院肿瘤二科,新疆乌鲁木齐830001

出　　处：《广西医科大学学报》2020年第8期1420-1428,共9页Journal of Guangxi Medical University

基　　金：国家自然科学基金-地区科学基金资助项目(No.8196140086)。

摘　　要：目的:探讨DH-332通过TRIM40/Raf-1/ERK1/2调节胃癌(GC)的发生以及机制研究。方法:首先,检测了新疆医科大学第四附属医院30例GC患者及相应的癌旁组织和GCMKN-45细胞及正常胃上皮GES-1细胞中TRIM40 mRNA及其蛋白的表达水平;然后构建了干扰TRIM40的MKN-45细胞系,将DH-332处理MKN-45细胞或干扰MKN-45细胞的TRIM40后,通过qPCR、Western blotting检测TRIM40、Raf-1、ERK1/2和细胞周期关键蛋白CyclinD1、CDK2及糖酵解相关蛋白GLUT1、PKM2、HKII、LDHA mRNA及其蛋白的表达,通过糖酵解相关试剂盒检测细胞内葡萄糖、乳酸及ATP的含量,CCK-8检测MKN-45细胞的增殖能力,最后通过集落形成、TranswellTM和划痕实验检测MKN-45细胞的侵袭和迁移能力。结果:与相邻的癌旁组织和GES-1细胞相比,TRIM40 mRNA和蛋白在GC组织及GC细胞中低表达;与MKN-45细胞相比,DH-332能够促进MKN-45细胞TRIM40 mRNA及其蛋白的表达,抑制Raf-1、CyclinD1、CDK、GLUT1、PKM2、HKII、LDHA mRNA及其蛋白的表达,抑制ERK1/2蛋白的表达;与TRIM40-shN对照组相比,干扰TRIM40后,Raf-1、CyclinD1、CDK、GLUT1、PKM2、HKII、LDHA mRNA及其蛋白的表达升高,ERK1/2蛋白的表达升高;与MKN-45细胞相比,DH-332能够抑制MKN-45细胞葡萄糖的摄取、乳酸和ATP的生成,抑制其增殖和侵袭迁移能力;与TRIM40-shN对照组相比,干扰TRIM40后,MKN-45细胞葡萄糖的摄取、乳酸和ATP的生成增加,增殖和侵袭迁移能力也升高。结论:DH-332通过促进GC细胞中TRIM40的表达,抑制Raf-1/ERK1/2通路的发生,并且抑制糖酵解的发生及细胞的增殖和侵袭迁移,减缓GC的进展。Objective:To explore the regulation and its mechanism of DH-332 on gastric cancer(GC)through TRIM40/Raf-1/ERK1/2.Methods:A total of 30 patients with gastric cancer were admitted to ourhospital.Firstly,the expression levels of TRIM40 mRNA and its protein in GC tissues,paracancerous tissues,GC MKN-45 cells and normal gastric epithelial GES-1 cells were detected.Then the MKN-45 cell line interfering with TRIM40 was constructed.After MKN-45 cells were treated with DH-332 or TRIM40 of MKN-45 cells were interfered,the expressions of TRIM40,Raf-1,ERK1/2,CyclinD1,CDK2 and glycolysis related proteins GLUT1,PKM2,HKII,LDHA mRNA and their proteins were detected by qPCR and Western blotting.The contents of glucose,lactic acid and ATP in cells were detected by glycolysis related kit.The proliferation ability of MKN-45 cells was detected by CCK-8,and the invasion and migration ability of MKN-45 cells was detected by colony formation,TranswellTM and scratch test.Results:Compared with adjacent paracancerous tissues and GES-1 cells,TRIM40 mRNA and protein were lowly expressed in GC tissues and GC cells.Compared with MKN-45 cells,DH-332 could promote the expression of TRIM40 mRNA and its protein in MKN-45 cells,inhibit the expression of Raf-1,CyclinD1,CDK,GLUT1,PKM2,HKII,LDHA mRNA and their proteins,and inhibit the expression of ERK1/2 protein.Compared with TRIM40-shN control group,the expression of Raf-1,CyclinD1,CDK,GLUT1,PKM2,HKII,LDHA mRNA and ERK1/2 protein was increased after interfering with TRIM40.Compared with MKN-45 cells,DH-332 could inhibit glucose uptake,lactic acid and ATP production,proliferation,invasion and migration of MKN-45 cells.Compared with TRIM40-shN control group,after interfering with TRIM40,glucose uptake,lactic acid and ATP production,proliferation,invasion and migration ability of MKN-45 cells were also increased.Conclusion:DH-332 slows down the progress of GC by promoting the expression of TRIM40 in GC cells,inhibiting the occurrence of Raf-1/ERK1/2 pathway,and inhibiting the occurrence of glycolysis a

关 键 词：胃癌 DH-332 TRIM40 Raf-1/ERK1/2 糖酵解 增殖和侵袭迁移 

分 类 号：R735.2[医药卫生—肿瘤]