CAJ | 학술논문

目的观察基质金属蛋白酶-9(MMP-9)在胃癌发生、发展中的作用；构建MMP-9基因沉默质粒；建立MMP-9基因沉默的BGC823细胞.方法采用免疫组织化学法在46例人胃癌新鲜标本中检测MMP-9的蛋白表达水平,采用化学合成的方法合成3条沉默MMP-9的寡聚核苷酸序列,插入到短发卡RNA (shRNA)表达载体pGenesil上,运用荧光显微镜、明胶酶电泳、逆转录-聚合酶链反应(RT-PCR)分别检测3条质粒的沉默效率；选取沉默效率最佳的1个质粒转染BGC823进行稳定细胞株的筛选及鉴定.结果 46例胃癌组织MMP-9蛋白阳性率为100％,其中57％(26/46)呈增强表达；筛选到1条MMP-9沉默最为显著的shRNA表达质粒；成功构建MMP-9基因沉默的BGC823胃癌细胞克隆.结论 MMP-9基因在胃癌发生、发展及浸润、转移过程中可能扮演重要角色.
목적 관찰기질금속단백매-9(MMP-9)재위암발생、발전중적작용；구건MMP-9기인침묵질립；건립MMP-9기인침묵적BGC823세포.방법 채용면역조직화학법재46례인위암신선표본중검측MMP-9적단백표체수평,채용화학합성적방법합성3조침묵MMP-9적과취핵감산서렬,삽입도단발잡RNA (shRNA)표체재체pGenesil상,운용형광현미경、명효매전영、역전록-취합매련반응(RT-PCR)분별검측3조질립적침묵효솔；선취침묵효솔최가적1개질립전염BGC823진행은정세포주적사선급감정.결과 46례위암조직MMP-9단백양성솔위100％,기중57％(26/46)정증강표체；사선도1조MMP-9침묵최위현저적shRNA표체질립；성공구건MMP-9기인침묵적BGC823위암세포극륭.결론 MMP-9기인재위암발생、발전급침윤、전이과정중가능분연중요각색.
Objective To investigate the roles of matrix metalloproteinase-9 (MMP-9) in gastric cancer development,to construct MMP-9 short hairpin RNA (shRNA) expression plasmids and to establish the gastric cancer cell line with silenced MMP-9 gene expression.Methods The expression level of MMP9 in 46 fresh specimens was detected by using immunohistochemistry.Three oligonucleotides which silenced MMP-9 gene expression were synthesized chemically and inserted into shRNA expression vector pGenesil to construct MMP-9 shRNA plasmids.Immunofluorescence,gelatin zymography and reverse transcription-polymerase chain reaction (RT-PCR) were applied to detect the MMP-9 silencing efficiency.The MMP-9 knockdown plasmid that had a high efficiency in silencing MMP-9 gene expression was chosen and transfected into BGC823 to establish the cell line with silenced MMP-9 gene expression.Results MMP-9 protein was expressed in 46 cases of gastric cancers,and strong expression was found in 57％ (26/46) of the gastric cancers.A shRNA expression plasmid with markedly silenced MMP-9 gene was screened out.BGC823 cell line with silenced MMP-9 gene was successfully constructed.Conclusion MMP-9 may play an important role in gastric cancer development,migration and invasion.