KLF13上调对骨肉瘤细胞侵袭、迁移及p-AKT表达的影响

  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=1 544.143、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。 

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

  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=1 544.143, 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.