KLF13上调对骨肉瘤细胞侵袭、迁移及p-AKT表达的影响  

Effects of the up-regulation of KLF13 on invasion,migration and p-AKT expression of osteosarcoma cells

作　　者：潘寒松 郭昌军[1] 梁一民[1] 梁剑伟 李祥[1] PAN Han-song;GUO Chang-jun;LIANG Yi-min;LIANG Jian-wei;LI Xiang(Department of Orthopaedics,Taizhou First People’s Hospital,Taizhou,Zhejiang 318020,China)

机构地区：[1]台州市第一人民医院骨科,浙江台州318020

出　　处：《中华全科医学》2021年第12期2031-2034,2097,共5页Chinese Journal of General Practice

基　　金：浙江省基础公益研究计划项目(LY20H070006);台州市社会发展类一般项目(1701KY48)。

摘　　要：目的研究Kruppel样因子13(KLF13)对骨肉瘤细胞(MG-63)侵袭、迁移的影响及该过程中p-AKT蛋白表达变化。方法将MG-63细胞按照随机数字表法分为对照组、NC组和LV-KLF13-OE组;显微镜观察KLF13慢病毒颗粒转染情况;实时荧光定量PCR(qRT-PCR)检测KLF13 mRNA表达情况;蛋白免疫印迹法检测KLF13、丝氨酸/苏氨酸蛋白激酶B(AKT)及磷酸化水平、E-钙黏蛋白(E-cadherin)和膜棕榈糖基化蛋白2(MPP-2)的蛋白表达情况;细胞计数试剂盒8(CCK-8)法检测细胞增殖情况;划痕实验检测迁移能力;小室法(Transwell)检测侵袭能力。结果MG-63细胞转染效率>75%;与对照组和NC组相比,LV-KLF13-OE组细胞KLF13 mRNA和蛋白、E-cadherin蛋白表达升高(F=1544.143、588.000、235.391,均P<0.001),MPP-2蛋白表达水平降低(F=260.053,P<0.001);LV-KLF13-OE组细胞划痕愈合率[(20.07±8.18)%]、侵袭数量[(89.29±15.04)个]、p-AKT蛋白表达(0.21±0.02)水平低于对照组[(71.61±6.02)%、(221.33±20.09)个、1.12±0.12]和NC组[(74.93±7.21)%、(212.69±25.47)个、1.09±0.08,F=109.781、76.815、226.896,均P<0.001]。结论KLF13过表达可能通过下调p-AKT水平,抑制人骨肉瘤细胞迁移和侵袭。Objective To study the effects of Kruppel-like factor 13(KLF13)on the invasion and migration of osteosarcoma cells(MG-63)and changes in p-AKT protein expression during the process.Methods MG-63 cells were divided into control group,NC group and LV-KLF13-OE group according to random number table method.The transfection of KLF13 recombinant lentivirus particles was observed under a microscope.The expression levels of KLF13 mRNA were detected by real-time fluorescent quantitative PCR.In addition,the protein expression levels of KLF13,serine/threonine protein kinase B(AKT)and its phosphorylation,E-cadherin and membrane palmitoylated protein 2(MPP-2)were detected by western blot.Proliferation was detected by the Cell Counting Kit-8(CCK-8),the migration ability was detected by a scratch test,and invasiveness was detected by the transwell method.Results The transfection efficiency of MG-63 cells was over 75%.Compared with those in the control group and NC group,the expression levels of KLF13 mRNA and protein and E-cadherin protein in the LV-KLF13-OE group were significantly higher(F=1544.143,588.000,235.391,all P<0.001),and the expression levels of MPP-2 protein were significantly lower(F=260.053,P<0.001).The scratch healing rate[(20.07±8.18)%],number of invasions(89.29±15.04)and p-AKT protein expression(0.21±0.02)levels in the LV-KLF13-OE group were significantly lower than those in the control group[(71.61±6.02)%,221.33±20.09 and 1.12±0.12]and NC group[(74.93±7.21)%,212.69±25.47 and 1.09±0.08;F=109.781,76.815,226.896,all P<0.001].Conclusion KLF13 overexpression may inhibit the migration and invasion of human osteosarcoma cells by down-regulating p-AKT expression.

关 键 词：Kruppel样因子13 骨肉瘤细胞 增殖 迁移 侵袭 丝氨酸/苏氨酸蛋白激酶B 

分 类 号：R738.1[医药卫生—肿瘤] R730.231[医药卫生—临床医学]