IGSF10通过抑制FAK/PI3K/AKT信号通路降低右美托咪定对乳腺癌促进作用的研究  

IGSF10 reduces dexmedetomidine promotion in breast cancer by inhibiting FAK/PI3K/AKT signaling pathway

作　　者：吴娜萍 张磊[1] 方琦[1] 王磊[1] WU Na-ping;ZHANG Lei;FANG Qi;WANG Lei(Department of Breast Surgery,Third Affiliated Hospital of SoochowUniversity,Changzhou 213000,China)

机构地区：[1]苏州大学附属第三医院乳腺外科,江苏常州213000

出　　处：《中华肿瘤防治杂志》2022年第14期1052-1061,共10页Chinese Journal of Cancer Prevention and Treatment

基　　金：常州市卫健委科技项目(QN202012)。

摘　　要：目的探讨免疫球蛋白超家族成员10(IGSF10)对右美托咪定(DEX)干预的乳腺癌细胞增殖、迁移、侵袭、上皮细胞-间充质转化(EMT)的影响及作用机制。方法以人正常乳腺上皮细胞MCF-10A,人乳腺癌细胞MCF-7、ZR-75-1、MDA-MB-231和MDA-MB-415为研究对象,定量逆转录聚合酶链反应(qRT-PCR)和蛋白质印迹法检测IGSF10在细胞中的表达。选取IGSF10表达最低的细胞株为后续实验的研究对象。分为空白对照组、过表达阴性对照组和IGSF10过表达组,用于研究IGSF10对乳腺癌细胞恶性生物学行为的影响;分为空白对照组、DEX组、DEX+过表达阴性对照组和DEX+IGSF10过表达组,用于研究IGSF10对DEX诱导的乳腺癌细胞的影响。IGSF10过表达质粒载体转染乳腺癌细胞;细胞计数盒8(CCK8)、克隆形成实验、划痕实验、Transwell小室实验分别检测IGSF10对乳腺癌细胞增殖、迁移和侵袭的影响;蛋白质印迹法检测IGSF10对增殖细胞核抗原(PCNA)、Ki-67、基质金属蛋白酶(MMP)-2、MMP-9、E-钙黏蛋白(E-cadberin)、N-钙黏蛋白(N-cadherin)、波形蛋白(Vimentin)、黏着斑激酶(FAK)、磷脂酰肌醇3-激酶(PI3K)和蛋白激酶B(AKT)表达的影响。结果与人正常乳腺上皮细胞MCF-10A相比,乳腺癌细胞系MCF-7(0.66±0.06和0.76±0.07)、ZR-75-1(0.60±0.04和0.69±0.05)、MDA-MB-231(0.31±0.04和0.38±0.06)和MDA-MB-415(0.39±0.03和0.43±0.05)中IGSF10 mRNA和蛋白表达水平降低(F值分别为105.80和66.41,均P<0.001),选择IGSF10表达最低的MDA-MB-231细胞株进行后续实验。与空白对照组和过表达阴性对照组相比,IGSF10过表达组24和48 h细胞活力(F_(24 h)=35.84,F_(48 h)=28.30,均P<0.001)、克隆数量(F=57.42,P<0.001)、细胞迁移和侵袭能力(F_(迁移)=67.81,F_(侵袭)=84.22,均P<0.001)降低,PCNA、Ki-67、MMP-2、MMP-9、N-cadherin、Vimentin、磷酸化(p-)FAK/FAK、p-PI3K/PI3K和p-AKT/AKT蛋白表达下调(F值分别为20.68、22.58、38.49、31.93、35.49、28.33、78.22、52.17和10.Objective To investigate the effects and mechanism of immunoglobulin superfamily member 10(IGSF10)on proliferation,migration,invasion and epithelial mesenchymal transition(EMT)of breast cancer cells treated with dexmedetomidine(DEX).Methods Human normal breast epithelial cells(MCF-10 A),human breast cancer cells(MCF-7,ZR-75-1,MDA-MB-231 and MDA-MB-415)were used for the study,the mRNA and protein expression levels in the cells were detected by quantitative reverse transcription polymerase chain reaction(qRT-PCR)and western blot.One cell line with the lowest IGSF10 expression level was selected for the following experiments,and cells were divided into blank control group,overexpression negative control group and IGSF10overexpression group to study the effect of IGSF10on malignant biological behavior of breast cancer cells;blank control group,DEX group,DEX+overexpression negative control group and DEX+IGSF10overexpression group to evaluate the effect of IGSF10on DEX induced breast cancer cells.IGSF10overexpression plasmid vector was transfected into breast cancer cells;the effects of IGSF10on proliferation,migration and invasion of breast cancer cells were evaluated by CCK8,colony formation assay,wound healing assay and transwell invasion assay,respectively;the effects of IGSF10on the expression of proliferating cell nuclear antigen(PCNA),Ki-67,matrix metalloproteinase(MMP)-2,MMP-9,E-cadherin,N-cadherin,Vimentin,focal adhesion kinase(FAK),phosphatidylinositol 3kinase(PI3K)and protein kinase B(AKT)were determined by western blot.Results Compared with human normal breast epithelial cell line MCF-10A,the mRNA and protein expression levels of IGSF10in breast cancer cell lines MCF-7(0.66±0.06,0.76±0.07),ZR-75-1(0.60±0.04,0.69±0.05),MDA-MB-231(0.31±0.04,0.38±0.06)and MDA-MB-415(0.39±0.03,0.43±0.05)were significantly reduced,and the Fvalues were 105.80and 66.41,both P<0.001.MDA-MB-231cell line with the lowest expression of IGSF10was selected for following experiments.Compared with the blank control group and the ov

关 键 词：乳腺癌 免疫球蛋白超家族成员10 右美托咪定 FAK/PI3K/AKT信号通路 增殖 侵袭 迁移 

分 类 号：R737.9[医药卫生—肿瘤]