HER2靶向载HSV-TK基因纳米泡诱导胃癌细胞凋亡的体外研究  

Apoptosis Induced by HER2 Targeted HSV-TK Gene Loaded Nanobubbles in Gastric Cancer Cells:An in vitro Study

作　　者：辛莹 周厚妊 张月[1] 刘慧[2] 黄驰 刘治军[1] XIN Ying;ZHOU Houren;ZHANG Yue;LIU Hui;HUANG Chi;LIU Zhijun(Department of Ultrasound,Shengjing Hospital of China Medical University,Shenyang 110004,China)

机构地区：[1]中国医科大学附属盛京医院超声科,辽宁沈阳110004 [2]沈阳药科大学药学院,辽宁沈阳110016

出　　处：《中国医学影像学杂志》2021年第2期106-110,116,共6页Chinese Journal of Medical Imaging

基　　金：国家自然科学基金(81801712)。

摘　　要：目的探讨HER2靶向载单纯疱疹病毒胸苷激酶(HSV-TK)基因/AGM-CBA的全氟己烷纳米泡(tHAHT NB)的体外寻靶能力及核定位效应,以及在超声介导纳米泡破坏技术下tHAHT NB抑制胃癌细胞增殖和诱导细胞凋亡的作用。材料与方法将实验分为MGC803组及NCI-N87组,通过激光共聚焦显微镜检测tHAHT NB的体外靶向情况,使用ImageJ软件半定量分析比较两组细胞周围累计荧光密度,分析NCI-N87组累计荧光密度与细胞数的相关性;通过激光共聚焦显微镜检测tHAHT NB对NCI-N87细胞的核定位效应;将实验分为tHAHT NB+US组、tHAHT NB组、tHER2 NB组、NB组、US组、正常细胞组及空白组,以MTT法观察细胞活性、Annexin V检测细胞凋亡情况。结果激光共聚焦显微镜下显示与MGC803组相比,NCI-N87组细胞周围可见明显红色荧光;半定量分析显示,与MGC803组相比,NCI-N87组累计荧光密度增高,且与细胞数目呈正相关(r=0.900,P<0.01);核定位实验显示NCI-N87组细胞核周围可见明显红色荧光。MTT法及Annexin V检测发现,与tHAHT NB组、tHER2 NB组、NB组及US组相比,tHAHT NB+US组细胞活性明显减低,细胞凋亡率显著增高。结论tHAHT NB可以靶向识别NCI-N87细胞,并具有核定位效应;联合超声介导纳米泡破坏技术能明显抑制NCI-N87细胞活性,并诱导细胞凋亡,为胃癌的非侵入性治疗提供了新思路。Purpose To investigate the targeting ability in vitro and nuclear localization effect of targeted HER2 AGM-CBA and HSV-TK gene-loaded perfluorohexane nanobubble(tHAHT NB),and analyze the inhibition cell proliferation and inducing cell apoptosis of tHAHT NB under ultrasound-targeted nanobubble destruction technology.Materials and Methods The experimental cells were divided into MGC803 group and NCI-N87 group,the in vitro targeting effect of tHAHT NB was detected by confocal laser microscope,ImageJ software was used to semi-quantitatively analyze and compare the cumulative fluorescence density of cells between the two groups,and the correlation between cumulative fluorescence density of the NCI-N87 group and cell number was analyzed;the nuclear localization effect of tHAHT NB on NCI-N87 cells was detected by laser confocal microscope;the experimental cells were divided into tHAHT NB+US group,tHAHT NB group,tHER2 NB group,NB group and US group,then MTT method was used to observe cell viability and Annexin V to detect cell apoptosis.Results Confocal laser microscope showed that compared with the MGC803 group,the NCI-N87 group had obvious red fluorescence signals around the cells;semi-quantitative analysis showed that compared with the MGC803 group,the cumulative fluorescence density of the NCI-N87 group increased,and it was positively correlated with the cell number(r=0.900,P<0.01);nuclear localization experiment showed that there was obvious red fluorescence around the nucleus in the NCI-N87 group.MTT assay and Annexin V detection showed that compared with tHAHT NB group,tHER2 NB group,NB group and US group,the cell activity of tHAHT NB+US group was significantly reduced,and the apoptosis rate was significantly increased.Conclusion tHAHT NB can target and recognize NCI-N87 cells,and has a nuclear localization effect;combined with ultrasound-targeted nanobubble destruction technology,tHAHT NB can improve genes transfection rate to significantly inhibit the proliferation of NCI-N87 cells and induce apoptosis,which pro

关 键 词：胃肿瘤 纳米粒子 超声检查 HSV-TK基因 核定位效应 细胞凋亡 体外研究 

分 类 号：R-33[医药卫生]