脂筏标记蛋白flotillin-2对人乳腺癌细胞迁移和侵袭及阿霉素化疗敏感性的影响  

Effect of flotillin-2,a lipid raft protein,on migration,invasion and adriamycin sensitivity of human breast cancer cells

作　　者：赖斯华 史晓娜 徐霞 谢闺娥 LAI Si-hua;SHI Xiao-na;XU Xia;XIE Gui-e(KingMed School of Laboratory Medicine,Guangzhou Medical University,Guangzhou,Guangdong 510182,China)

机构地区：[1]广州医科大学金域检验学院,广东广州510182

出　　处：《热带医学杂志》2021年第11期1375-1378,1395,F0002,共6页Journal of Tropical Medicine

基　　金：广东省自然科学基金(2017A030313906);广州医科大学大学生实验室开放项目(2018-22)。

摘　　要：目的研究脂筏标记蛋白flotillin-2(FLOT2)在人乳腺癌细胞迁移、侵袭及阿霉素化疗敏感性调控中的生物学作用。方法利用课题组前期构建的下调FLOT2表达的MCF-7和MDA-MB-231稳定细胞系,用软琼脂细胞集落培养法检测FLOT2对乳腺癌细胞克隆形成的影响;划痕试验和Transwell小室迁移试验、Transwell小室侵袭试验分析FLOT2对乳腺癌细胞运动爬行能力、侵袭能力的作用;用MTT比色法及Annexin V荧光染色分别检测FLOT2与乳腺癌细胞对阿霉素敏感性及抗凋亡能力的关联作用。结果软琼脂集落培养显示沉默FLOT2的乳腺癌细胞在软琼脂中形成的细胞集落显著少于对照细胞,差异有统计学意义(P<0.05);划痕试验中沉默FLOT2的乳腺癌细胞运动距离较对照细胞显著减少,差异有统计学意义(P<0.05);沉默FLOT2的乳腺癌细胞穿过未铺基质胶(matrigel)和铺了matrigel小室底膜的细胞数量均显著少于对照细胞,差异有统计学意义(P<0.05)。阿霉素对对照MCF-7和MDA-MB-231作用48 h的IC50分别为1.14和0.90μmol/L;而对沉默FLOT2的MCF-7和MDA-MB-231的IC50分别降低为0.47和0.36μmol/L。阿霉素作用后沉默FLOT2的乳腺癌细胞凋亡比率显著高于对照细胞,差异有统计学意义(P<0.05)。结论 FLOT2在促进乳腺癌细胞迁移、侵袭中起重要作用,沉默FLOT2能增强乳腺癌细胞对阿霉素的敏感性。Objective To investigate the effect of the lipid raft protein flotillin-2(FLOT2)on migration,invasion,and adriamycin sensitivity of human breast cancer cells.Methods In FLOT2-knockdown MCF-7 and MDA-MB-231 stable cell lines,soft agar colony formation assay was conducted to analyze the effect of FLOT2 on colony formation ability.The function of FLOT2 on migration and invasion was evaluated by wound healing assay,transwell migration assay,and transwell invasion assay.The influence of FLOT2 on adriamycin sensitivity and apoptosis resistance of breast cancer cells was analyzed by MTT assay and Annexin V staining,respectively.Results Soft agar colony formation assay revealed that the number of cellular colonies was significantly decreased in FLOT2-knockdown breast cancer cells,as compared with the control cells(P<0.05).Wound healing assay showed that the distance of wound healing of FLOT2-knockdown breast cancer cells was significantly shorter than that of the control cells(P<0.05).The average number of migrated or invaded cells crossing the transwell membranes without or with matrigel was significantly decreased in FLOT2-knockdown breast cancer cells,as compared with the control cells(P<0.05).Upon treatment of adriamycin for 48 h,calculated IC50 values,i.e.,concentrations of adriamycin required for decreasing the growth rate by 50%,were 1.14 μmol/L for MCF-7 and 0.90 μmol/L for MDA-MB-231.For FLOT2-knockdown MCF-7 and MDA-MB-231 cells,IC50 values of adriamycin were 0.47 and 0.36μmol/L,respectively.After treatment with adriamycin,the apoptotic rate of FLOT2-knockdown breast cancer cells was significantly higher than that of the control cells(P<0.05).Conclusion FLOT2 might play an important role in promoting cell migration and invasion,and FLOT2 downreglution could enhance the sensitivity of breast cancer cells to adriamycin.

关 键 词：Flotillin-2 乳腺癌 迁移 侵袭 阿霉素 

分 类 号：R737.9[医药卫生—肿瘤]