CAJ | 학술논문

目的：研究RNA干扰（ RNA interference ，RNAi） MMP-9基因对THP-1细胞增殖的抑制作用。探讨RNA干扰MMP-9基因在白血病治疗中的应用。方法：设计合成针对MMP-9基因的小干扰RNA（ small interfering RNA，siRNA），转染THP-1细胞，采用RT-PCR法测定MMP-9 mRNA的表达，Western blot测定MMP-9蛋白的表达，MTT和台盼蓝染色观察RNAi后对THP-1细胞生长的影响，显微镜下观察细胞形态改变。结果：siRNA转染组THP-1细胞MMP-9 mRNA及蛋白表达受到抑制，显著低于空白对照组和阴性对照组。 MTT及台盼蓝染色结果显示，siRNA组转染后48 h、72 h，THP-1细胞的增殖能力明显下降，细胞生长明显受到抑制，细胞活率明显低于对照组（ P＜0.05）。倒置显微镜下发现，对照组细胞呈半贴壁生长，细胞轮廓清楚，形态均一，生长旺盛。而siRNA组细胞生长呈受抑制状态，Wright染色后观察，siRNA组细胞形态有明显改变，大部分细胞出现凋亡的形态学变化。结论：靶向MMP-9的siRNA 不仅能特异性降低 MMP-9 mRNA 及蛋白的表达，且能有效抑制THP-1细胞的增殖，促进THP-1细胞凋亡。
목적：연구RNA간우（ RNA interference ，RNAi） MMP-9기인대THP-1세포증식적억제작용。탐토RNA간우MMP-9기인재백혈병치료중적응용。방법：설계합성침대MMP-9기인적소간우RNA（ small interfering RNA，siRNA），전염THP-1세포，채용RT-PCR법측정MMP-9 mRNA적표체，Western blot측정MMP-9단백적표체，MTT화태반람염색관찰RNAi후대THP-1세포생장적영향，현미경하관찰세포형태개변。결과：siRNA전염조THP-1세포MMP-9 mRNA급단백표체수도억제，현저저우공백대조조화음성대조조。 MTT급태반람염색결과현시，siRNA조전염후48 h、72 h，THP-1세포적증식능력명현하강，세포생장명현수도억제，세포활솔명현저우대조조（ P＜0.05）。도치현미경하발현，대조조세포정반첩벽생장，세포륜곽청초，형태균일，생장왕성。이siRNA조세포생장정수억제상태，Wright염색후관찰，siRNA조세포형태유명현개변，대부분세포출현조망적형태학변화。결론：파향MMP-9적siRNA 불부능특이성강저 MMP-9 mRNA 급단백적표체，차능유효억제THP-1세포적증식，촉진THP-1세포조망。
To study the inhibitory effect of RNA interference (RNAi) on MMP-9 gene expression in THP-1 cell line.To investigate the application of RNAi on the therapy of leukemia.Methods:Small interfering RNA ( siRNA) for MMP-9 gene was designed and transfected into THP-1 cells.MMP-9 mRNA expression was assessed by RT-PCR, and MMP-9 protein expression was tested by Western blot.MTT and trypan blue staining were used to observe the effect on the proliferation of THP-1 cells after RNAi.The changes in cell morphology were observed under the microscope.Results:The expressions of MMP-9 mRNA and protein were inhibited in THP-1 MMP-9 siRNA-transfected cells ,significantly lower than those of control cells.The results of MTT and trypan blue staining in-dicated that the proliferation ability of THP-1 cells obviously decreased after siRNA-transfected 48h and 72h.The growth of cells was in-hibited and the cells survival rate was significantly lower than that of control group ( P<0.05 ).The cells of control groups grew semi-quote wall under inverted microscope.The outline of cells was clear and the shape was uniform.The cells grew vigorously.While the growth of cells in siRNA group was inhibited.The morphology of siRNA group cells changed obviously by the Wright staining.Most cells expressed changes of apoptosis.Conclusion: siRNA for MMP-9 gene can not only reduce the expressions of MMP-9 mRNA and protein,but also inhibit the proliferation and induce apoptosis of THP-1 cells.