敲低PHF6对三阴性乳腺癌细胞迁移能力的影响探讨  

Effect of PHF6 knockdown on migration of triple negative breast cancer cells

作　　者：崔敬轶 刘玉婷 包欣玥 富春杭 唐宇 陈竹 徐亚亚 李丹 CUI Jing-yi;LIU Yu-ting;BAO Xin-yue(School of Basic Medicine,Shenyang Medical College,Shenyang 110034,China)

机构地区：[1]沈阳医学院基础医学院,110034

出　　处：《中国现代药物应用》2024年第14期176-180,共5页Chinese Journal of Modern Drug Application

基　　金：国家自然科学基金青年科学基金项目(项目编号:82103188);辽宁省教育厅科学研究经费项目(面上项目)(项目编号:LJKZ1148);辽宁省科学技术计划项目(面上项目)(项目编号:2023-MS-325);沈阳医学院大学生科研课题(项目编号:20219007)。

摘　　要：目的探讨植物同源结构域指蛋白6(PHF6)对三阴性乳腺癌MDA-MB-231细胞迁移能力的影响。方法通过癌症基因组图谱(TCGA)中的数据分析PHF6在各类乳腺癌组织[Luminal型、人表皮生长因子受体-2(HER2)高表达型和三阴性乳腺癌]中的表达情况及其表达对患者生存率的影响。利用siRNA干扰技术在三阴性乳腺癌细胞MDA-MB-231中敲低PHF6,通过划痕实验检测敲低PHF6后细胞迁移能力的变化。采用实时荧光定量逆转录聚合酶链式反应(RT-PCR)实验检测乳腺癌细胞中上皮细胞-间充质转化(EMT)相关基因的表达情况。结果PHF6在各类乳腺癌中(Luminal型、HER2高表达型和三阴性乳腺癌)均出现了明显过表达现象,三阴性乳腺癌中表达最高,且高表达PHF6降低了乳腺癌患者的存活率。用siNC、siPHF6转染MDA-MB-231乳腺癌细胞,24 h后RT-PCR结果显示,与siNC组细胞中PHF6的RNA表达水平(1.033±0.09238)相比,siPHF6组细胞中PHF6的RNA表达水平(0.1054±0.006194)明显下调(P<0.001)。划痕结果显示,与siNC组划痕后6、18、24 h划痕间隙[(433.82±18.28)、(217.87±18.43)、(86.96±39.72)μm]相比,siPHF6组在划痕后6、18、24 h划痕间隙[(639.86±30.63)、(464.25±21.41)、(248.55±20.17)μm]均较大(P<0.001、<0.001、<0.01),愈合较慢。RT-PCR结果显示,与siNC组EMT相关基因CDH2、波形蛋白(VIM)的表达[(1.01±0.056)、(1.01±0.045)]相比,在MDA-MB-231细胞中敲低PHF6,siPHF6组CDH2、VIM表达[(0.25±0.036)、(0.632±0.054)]均降低(P<0.001、<0.001)。结论敲低PHF6能抑制三阴性乳腺癌MDA-MB-231细胞的迁移能力。Objective To explore the effect of plant homeodomain finger protein 6(PHF6)on the migration of triple negative breast cancer cell line MDA-MB-231.Methods Based on the data in the cancer genome atlas(TCGA),the expression of PHF6 in various breast cancer[Luminal type,human epidermal growth factor receptor(HER)2+and triple negative breast cancer]and its impact on the survival rate of patients were analyzed.siRNA interference was used to knock down PHF6 in triple negative breast cancer cells MDA-MB-231,and the changes of cell migration after PHF6 knockdown were detected by scratch test.Real-time polymerase chain reaction(RT-PCR)was used to detect the expression of epithelial-mesenchymal transition(EMT)related genes in breast cancer cells.Results PHF6 was significantly overexpressed in various breast cancer(Luminal type,HER2+and triple negative breast cancer),with the highest expression in triple negative breast cancer and high expression of PHF6 reduces the survival rate of breast cancer patients.MDA-MB-231 breast cancer cells were transfected with siNC and siPHF6,and the results of inverse RT-PCR after 24 h showed that the RNA expression level of PHF6 was significantly down-regulated in cells of siPHF6 group(0.1054±0.006194)compared with that in cells of siNC group(1.033±0.09238)(P<0.001).The scratch tests showed that compared with the scratch gaps[(433.82±18.28),(217.87±18.43),(86.96±39.72)μm]in the siNC group at 6,18 and 24 h after scratching,the scratch gaps were larger in the siPHF6 group at 6,18 and 24 h after scratching[(639.86±30.63),(464.25±21.41),(248.55±20.17)μm](P<0.001,<0.001,<0.01)and healed more slowly.RT-PCR results showed that compared with the expression of EMT-related genes CDH2 and vimtentin(VIM)in the siNC group[(1.01±0.056),(1.01±0.045)],knockdown of PHF6 in MDA-MB-231 cells,the expression of CDH2 and VIM in the siPHF6 group[(0.25±0.036),(0.632±0.054)]were all decreased(P<0.001,<0.001).Conclusion PHF6 knockdown can inhibit the migration of triple negative breast cancer MDA-MB-231 c

关 键 词：植物同源结构域指蛋白6 三阴性乳腺癌细胞 迁移 上皮细胞-间充质转化 

分 类 号：R737.9[医药卫生—肿瘤]