低频超声辐照联合自噬抑制抗小鼠4T1乳腺癌的实验研究  

Antitumor effect of low-frequency ultrasound combined with autophagy blockade in the mouse model of 4T1 breast cancer

作　　者：王潇旋 商蒙蒙 孙霄[1] 郭鲁[1] 时丹丹[1] 李杰[1] Wang Xiaoxuan;Shang Mengmeng;Sun Xiao;Guo Lu;Shi Dandan;Li Jie(Department of Ultrasound,Qilu Hospital of Shandong University,Jinan 250012,China)

机构地区：[1]山东大学齐鲁医院超声科,济南250012

出　　处：《中华超声影像学杂志》2022年第3期260-266,共7页Chinese Journal of Ultrasonography

基　　金：国家自然科学基金面上项目(82071937,81771843)。

摘　　要：目的探讨低频超声辐照联合自噬抑制对小鼠4T1乳腺癌生长和侵袭的影响。方法取对数生长期的4T1细胞,分为对照组、超声组、羟氯喹组、超声联合羟氯喹组。蛋白质免疫印迹检测细胞LC3、p62、MMP-9的表达水平;透射电镜观察细胞内自噬体的形成;EdU与CCK-8测定细胞的增殖与细胞活力;Transwell法测定细胞侵袭力;流式细胞术检测对细胞凋亡的影响。构建4T1乳腺癌BALB/c小鼠移植瘤模型,分为对照组、超声组、羟氯喹组、超声联合羟氯喹组,每组5只,每2 d测量肿瘤体积及小鼠体重。结果超声联合羟氯喹组LC3-Ⅱ与p62的蛋白水平表达增加,透射电镜可见细胞内明显自噬体积累;在细胞实验中,与其他三组相比,超声联合羟氯喹组的生长抑制作用更强,细胞增殖率显著下降,MMP-9的表达降低,并能明显抑制侵袭(均P<0.050);在体内实验中,与对照组相比,各组小鼠肿瘤生长速度下降(均P<0.050),超声联合羟氯喹组较超声组和羟氯喹组治疗效果更优(均P<0.001)。结论低频超声辐照联合羟氯喹能够协同抑制肿瘤细胞增殖,促进细胞凋亡,明显减缓4T1荷瘤小鼠肿瘤生长。Objective To investigate the effect of low-frequency ultrasound combined with hydroxychloroquine(HCQ)on the growth and invasion of 4T1 breast cancer in mice.Methods 4T1 cells in logarithmic growth phase were divided into 4 groups:control group,ultrasound group,HCQ group and ultrasound combined with HCQ group.Western blot was performed to detect the effects of ultrasound combined with HCQ on the protein expression levels of autophagy-related proteins LC3,p62 and matrix metalloproteinase 9(MMP-9).Transmission electron microscopy was used to observe the formation of autophagic vesicles.The cell proliferation and cell viability were determined by EdU and CCK-8.Transwell assay was performed to detect the effect of ultrasound combined with HCQ on the invasive ability of 4T1 cells.Flow cytometry was performed to detect the effect of ultrasound combined with HCQ on the apoptosis of 4T1 cells.The transplantation tumor model of 4T1 breast cancer in BALB/c mice was constructed.The mice were randomly divided into 4 groups(n=5 for each group),including control group,ultrasound group,HCQ group,ultrasound combined with HCQ group.The tumor volume and mice body weight were evaluated and measured in each group every 2 days.Results The expression of LC3-Ⅱand p62 protein levels increased in the ultrasound combined with HCQ group,and intracellular autophagosome accumulation was evident by transmission electron microscopy.In cellular experiments,compared with the other groups,the ultrasound combined with HCQ group showed stronger growth inhibition,significantly decreased cell proliferation rate,decreased expression of MMP-9,and significantly inhibited invasion(all P<0.050).In the in vivo experiments,compared with the control group,the tumor growth rate of all 3 inter vention groups of mice decreased(all P<0.050),and the ultrasound combined with HCQ group had better therapeutic effects than the ultrasound and HCQ groups.The treatment effect of ultrasound combined with HCQ group was better than that of ultrasound and HCQ groups(all P<

关 键 词：超声处理 自噬 侵袭 增殖 凋亡 

分 类 号：R737.9[医药卫生—肿瘤]