细丝蛋白A对人胃癌细胞MKN28增殖及侵袭性的影响

Effect of filamin A on proliferation and invasion of human gastric cancer MKN28 cells

目的 探讨细丝蛋白A（FLNa）基因对人胃癌细胞MKN28体外增殖和侵袭的影响及机制.方法 通过脂质体介导的方式将重组质粒载体pcDNA3.1/VS-His-TOPO/FLNa转染到MKN28细胞中,Western blot检测转染后MKN28细胞中FLNa蛋白表达,噻唑蓝（MTT）法检测细胞的体外增殖能力,Transwell法检测细胞的侵袭能力.Western blot检测转染后细胞及对照细胞中细胞周期蛋白D1（Cyclin D1）及基质金属蛋白酶-9（MMP-9）的表达.结果 FLNa基因转染到MKN28细胞后,FLNa蛋白水平明显增加（P＜0.05）,验证转染成功.MTT结果显示,FLNa转染组在第3、4天的增殖能力明显低于对照组（0.45±0.04比0.62 ±0.02、0.65±0.03比0.81±0.03,P＜0.05）.Transwell细胞侵袭实验显示,FLNa转染组中穿透的细胞数明显少于空转组和对照组[（266±12）个比（405±16）个比（426±13）个,P＜0.05].FLNa转染到细胞后,明显抑制了Cyclin D1和MMP-9的蛋白表达（P＜0.05）.结论 FLNa基因具有抑制人胃癌MKN28细胞体外增殖和侵袭作用,其作用机制涉及Cyclin D1和MMP-9的表达下调.

Objective To explore the effect of filamin A （FLNa） gene on the proliferation and invasion of human gastric cancer MKN28 cells and to elucidate the possible mechanism.Methods Recombinant vector plasmid of pcDNA3.1/V5-His-TOPO/FLNa was transfected into MKN28 cells by lipofectAMINE.The expression of FLNa protein in MKN28/FLNa cells was detected by Western blotting.The proliferation rate of MKN28/FLNa cells was determined by methyl thiazol tetrazolium （MTT） assay.The invasion ability of MKN28/FLNa cells was determined by Transwell invasion assay.The expression of Cyclin D1 and matrix metalloproteinase-9 （MMP-9） protein in MKN28/FLNa cells was detected by Western blotting.Results The expression of FLNa protein was enhanced in MKN28 cells transfected with FLNa （P 〈 0.05）.Compared to control group,the proliferation of MKN28/FLNa cells on the 3rd day and 4th day was significantly inhibited （0.45 ±0.04 vs.0.62 ±0.02,and 0.65 ±0.03 vs.0.81 ±0.03,P 〈 0.05）.Compared to control group,the invasion ability of MKN28/FLNa cells was significantly decreased [（266 ± 12） cells vs.（405 ± 16） cells vs.（426 ± 13） cells,P 〈 0.05].The expression levels of Cyclin D1 and MMP-9 protein in MKN28/FLNa cells were downregulated （P 〈 0.05）.Conclusion The FLNa gene could inhibit the proliferation and invasion of human gastric cancer MKN28 cells,which may be associated with down-regulation of Cyclin D1 and MMP-9 expression.