CAJ | 학술논문

目的:探讨NGX6基因对结肠癌Wnt信号转导通路β-catenin/TCF/LEF转录活化的影响,明确NGX6在Wnt/β-catenin信号转导通路中的作用机制.方法:(1)构建β-catenin野生型真核表达载体pcDNA3.1(+)-β-catenin(WT),(2)转染外源性pcDNA3.1(+)-β-catenin(WT)及pCMV-myc-NGX6于COS-7细胞,TCF-4荧光素酶报告质粒检测转染NGX6前后TCF-4的转录活性.(3)无外源性β-catenin,pCMV-myc-NGX6单独转染COS-7细胞及结肠癌SW620细胞,TCF-4荧光素酶报告质粒检测TCF-4的转录活性,并采用Western blot的方法检测两组细胞核内β-catenin及TCF-4的表达.结果:(1)成功构建真核表达载体pcDNA3.1(+)-β-catenin(WT);(2)pcDNA3.1(+)-β-catenin(WT)与pCMV-myc-NGX6共转染COS-7细胞较pcDNA3.1(+)-β-catenin(WT)单独转染时TCF-4荧光素酶活性明显下降(P＜0.05);(3)无外源性β-catenin,pCMV-myc-NGX6转染COS-7及SW620细胞后TCF-4荧光素酶活性较空白质粒转染组下降(P＜0.05);(4)NGX6转染COS-7及SW620后核内β-catenin及TCF-4的表达下降.结论:NGX6对结肠癌中Wnt/β-catenin通路的β-catenin/TCF/LEF转录活化具有负性调节作用,其作用机制可能与NGX6抑制β-catenin的核转位有关.
목적:탐토NGX6기인대결장암Wnt신호전도통로β-catenin/TCF/LEF전록활화적영향,명학NGX6재Wnt/β-catenin신호전도통로중적작용궤제.방법:(1)구건β-catenin야생형진핵표체재체pcDNA3.1(+)-β-catenin(WT),(2)전염외원성pcDNA3.1(+)-β-catenin(WT)급pCMV-myc-NGX6우COS-7세포,TCF-4형광소매보고질립검측전염NGX6전후TCF-4적전록활성.(3)무외원성β-catenin,pCMV-myc-NGX6단독전염COS-7세포급결장암SW620세포,TCF-4형광소매보고질립검측TCF-4적전록활성,병채용Western blot적방법검측량조세포핵내β-catenin급TCF-4적표체.결과:(1)성공구건진핵표체재체pcDNA3.1(+)-β-catenin(WT);(2)pcDNA3.1(+)-β-catenin(WT)여pCMV-myc-NGX6공전염COS-7세포교pcDNA3.1(+)-β-catenin(WT)단독전염시TCF-4형광소매활성명현하강(P＜0.05);(3)무외원성β-catenin,pCMV-myc-NGX6전염COS-7급SW620세포후TCF-4형광소매활성교공백질립전염조하강(P＜0.05);(4)NGX6전염COS-7급SW620후핵내β-catenin급TCF-4적표체하강.결론:NGX6대결장암중Wnt/β-catenin통로적β-catenin/TCF/LEF전록활화구유부성조절작용,기작용궤제가능여NGX6억제β-catenin적핵전위유관.