慢病毒转染敲低HDAC1表达对胃癌干细胞干性增殖、迁移及侵袭的影响及其机制研究  

Effect of lentivirus transfected low HDAC1 expression on the dry proliferation,migration and invasion of gastric cancer stem cells and its mechanism

作　　者：刘小娟[1] 刘复娜[1] 张艳芳[1] 陈策[1] LIU Xiaojuan,LIU Funa,ZHANG Yanfang,CHEN Ce(Department of Digestive,General Hospital of North China Petroleum Administration Bureau, Renqiu 062552, China)

机构地区：[1]华北石油管理局总医院消化科,河北任丘062552

出　　处：《疑难病杂志》2018年第7期715-718,728,共5页Chinese Journal of Difficult and Complicated Cases

摘　　要：目的通过慢病毒转染敲低胃癌干细胞观察HDAC1表达对人类胃癌干细胞干性增殖、迁移及侵袭的影响以及对胃癌干细胞干性相关基因表达的影响,进一步探讨HDAC1低表达对人类胃癌干细胞增殖分化的影响及机制。方法2017年7—10月于华北石油管理局总医院细胞实验室进行。通过慢病毒转染将细胞株分为实验组对照组。以Nanog为胃癌细胞干性标志物,流式细胞术筛选出胃癌干细胞。通过慢病毒转染技术干扰胃癌干细胞中HDAC1表达,实时定量PCR(qRT-PCR)和Western Blot方法检测干扰效率。采用MTT比色法和划痕试验分别观察2组细胞增殖和迁移能力的差异;通过Transwell实验观察2组细胞侵袭能力的差异;采用qRT-PCR和Western Blot方法检测2组细胞干性相关基因表达。结果(1)实验组胃癌干细胞的增殖能力在96 h、120 h,均显著低于对照组(t=5.471、7.251,P均<0.001)。(2)通过细胞划痕实验观察到实验组胃癌干细胞的24 h划痕愈合速度较对照组慢,HDAC1的低表达降低胃癌干细胞的迁移能力,2组比较差异具有统计学意义[(20.23±5.46)个vs.(39.96±3.48)个,t=6.813,P=0.001]。(3)Transwell实验显示,实验组胃癌干细胞的侵袭能力显著低于对照组,HDAC1低表达降低胃癌干细胞的迁移能力,2组穿膜细胞数差异具有统计学意义[(21.51±4.32)个vs.(62.63±4.21)个,t=15.242,P=0.000]。实验组细胞干性相关基因Nanog、BMil、c-Myc表达均显著低于对照组(0.23±0.06 vs.1.00±0.01,t=28.306,P=0.000;0.64±0.11 vs.1.00±0.01,t=7.288,0.001;0.48±0.09 vs.1.00±0.01=12.841,0.000)。结论HDAC1低表达可降低胃癌干细胞的增殖、迁移及侵袭能力;HDAC1低表达下调BMi1基因表达可能是其降低胃癌干细胞的增殖、迁移及侵袭的机制之一。Objective To investigate the effect of HDAC1 expression on sternness proliferation, migration and invasion of human gastric cancer and the expression of stem cell related genes in human gastric cancer were observed by lentivirus transfected to low gastric cancer stem cells, and the effect and mechanism of HDAC1 low expression on the proliferation and differentiation of human gastric cancer stem cells were further explored. Methods From July 2017 to October 2017, at the cell laboratory of the General Hospital of the North China Petroleum Administration, the GCSCs were selected by flow cytometry with Nanog as marker. The expression of HDAC1 in GCSCs was down-regulated by lenti viral vector and detected by realtime quantitative reverse transcription PCR（qRT PCR） and Western Blot. The difference of Proliferation and immigration between two groups were evaluated by MTT assays and scratch test respectively; The difference of invasion between two groups were evaluated by transwell assays; The expression of sternness related genes was detected by qRT PCR and Western Blot. Results The proliferation（t = 7. 251. P=0.001） and immigration（t=6.813, P=0.001）in the experimental group were obviously lower than control group. The expression of sternness related genes in the experimental group were obviously lower than control group（ t=28. 306, P = 0.000; t =7.288. P = 0. 001; t = 12. 841, P = 0. 000）. Conclusion The low expression of HDAC1 can decrease proliferation, differentiation and invasion in gastric cancer stem cells. The low expression of HDAC1 down-regulate the expression of BMI1, which was thought to be one of the mechanism that HDAC1 affect proliferation and differentiation in human gastric cancer stem cells.

关 键 词：HDAC1 胃癌干细胞 增殖 迁移 侵袭 NANOG BMI1 C-Myc 

分 类 号：R735.2[医药卫生—肿瘤]