摘要
目的:探究lncRNA MCF2L-AS1对乳腺癌细胞增殖、迁移和侵袭的影响与机制。方法:qRT-PCR检测正常人乳腺上皮细胞系MCF-10A、乳腺癌细胞系(MDA-MB-415、MCF7、SKBR3、MDA-MB-231)中lncRNA MCF2L-AS1、miR-138-5p水平;将si-NC、si-MCF2L-AS1、si-MCF2L-AS1+inhibitor NC、si-MCF2L-AS1+miR-138-5p inhibitor分别转染至MDA-MB-415细胞并命名为si-NC组、si-MCF2L-AS1组、si-MCF2L-AS1+inhibitor NC组、si-MCF2L-AS1+miR-138-5p inhibitor组,未做任何处理的MDA-MB-415细胞记为NC组。qRT-PCR检测MDA-MB-415细胞中lncRNA MCF2L-AS1、miR-138-5p表达;MTT法检测MDA-MB-415细胞增殖情况;流式细胞术检测MDA-MB-415细胞凋亡率;Transwell检测MDA-MB-415细胞侵袭、迁移数量;Western blot检测MDA-MB-415细胞E-cadherin、Vimentin、N-cadherin、AnxA2蛋白水平;双荧光素酶报告基因实验验证lncRNA MCF2L-AS1、miR-138-5p、AnxA2的关系;小鼠异种移植实验检测肿瘤生长。结果:与MCF-10A细胞相比,MDA-MB-415、MCF7、SKBR3、MDA-MB-231细胞中lncRNA MCF2L-AS1、AnxA2水平显著上调(P<0.05),miR-138-5p表达显著下调(P<0.05)。与NC组、si-NC组相比,si-MCF2L-AS1组MDA-MB-415细胞OD 560 nm值、迁移、侵袭细胞数量、lncRNA MCF2L-AS1表达、N-cadherin、Vimentin蛋白水平、小鼠肿瘤体积、肿瘤质量显著下降(P<0.05),MDA-MB-415细胞凋亡率、miR-138-5p表达、E-cadherin蛋白水平显著升高(P<0.05);而抑制miR-138-5p表达减弱了沉默lncRNA MCF2L-AS1抑制MDA-MB-415细胞增殖、迁移、侵袭和肿瘤生长的效果;lncRNA MCF2L-AS1负向调控miR-138-5p/AnxA2轴。结论:沉默lncRNA MCF2L-AS1可能通过上调miR-138-5p来下调AnxA2的表达,进而抑制乳腺癌MDA-MB-415细胞增殖、迁移和侵袭。
Objective:To explore the effects and mechanisms of lncRNA MCF2L-AS1 on proliferation,migration and invasion of breast cancer cells.Methods:The levels of lncRNA MCF2L-AS1 and miR-138-5p in normal human breast epithelial cell line MCF-10A and breast cancer cell lines(MDA-MB-415,MCF7,SKBR3,MDA-MB-231)were detected by qRT-PCR.si-NC,si-MCF2L-AS1,si-MCF2L-AS1+inhibitor NC,and si-MCF2L-AS1+miR-138-5p inhibitor were transfected into MDA-MB-415 cells respectively,and named as si-NC group,si-MCF2L-AS1 group,si-MCF2L-AS1+inhibitor NC group,and si-MCF2L-AS1+miR-138-5p inhibitor group.MDA-MB-415 cells without any treatment were recorded as NC group.qRT-PCR was applied to detect the expression of lncRNA MCF2L-AS1 and miR-138-5p in MDA-MB-415 cells.MTT method was applied to detect the proliferation of MDA-MB-415 cells.Flow cytometry was applied to detect the apoptosis rate of MDA-MB-415 cells.Transwell was applied to detect the invasion and migration of MDA-MB-415 cells.Western blot was applied to detect the levels of E-cadherin,Vimentin,N-cadherin,and AnxA2 proteins in MDA-MB-415 cells.Double luciferase reporter gene experiment was applied to verify the relationship between lncRNA MCF2L-AS1,miR-138-5p and AnxA2.Mouse xenograft experiment was applied to detect tumor growth.Results:Compared with MCF-10A cells,the levels of lncRNA MCF2L-AS1 and AnxA2 in MDA-MB-415,MCF7,SKBR3 and MDA-MB-231 cells were obviously up-regulated(P<0.05),while the expression of miR-138-5p was obviously down-regulated(P<0.05).Compared with the NC group and si-NC group,the OD 560 nm value,migration and invasion cells numbers,lncRNA MCF2L-AS1 expression,N-cadherin and Vimentin protein levels,mouse tumor volume,and tumor quality of MDA-MB-415 cells in the si-MCF2L-AS1 group obviously decreased(P<0.05),the apoptosis rate,miR-138-5p expression,and E-cadherin protein level of MDA-MB-415 cells were obviously increased(P<0.05).The inhibiting the expression of miR-138-5p weakened the inhibitory effect of silencing lncRNA MCF2L-AS1 on the proliferation,migration,i
作者
王金礼
陈登峰
WANG Jinli;CHEN Dengfeng(Breast Department,Jingzhou Central Hospital,Hubei Jingzhou 434020,China)
出处
《现代肿瘤医学》
CAS
2024年第2期240-248,共9页
Journal of Modern Oncology
基金
荆州市医疗卫生科技计划项目(指导性计划)(编号:2021HC10)。
