CAJ | 학술논문

目的:探讨靶向多药耐药蛋白-1 (MDR1)基因的RNA干扰(RNAi)对结肠癌细胞MDR1/P-gp依赖的多药耐药性的稳定逆转作用.方法:分别构建含编码#4029 MDR1 siRNA和#4123 MDR1 siRNA的质粒载体,并转染COLO 320DM结肠癌多药耐药细胞,采用G418筛选克隆细胞,经实时定量RT-PCR及Western blotting鉴定阳性克隆细胞.MTT法检测细胞活力并计算各抗肿瘤药的IC50.流式细胞术检测细胞周期并计算PI/AI值.流式细胞仪测定细胞内adriamycin药物累积浓度.结果:阳性克隆细胞(clone #4029和clone #4123)的MDR1 mRNA和P-gp的表达均被抑制.COLO 320DM结肠癌亲本细胞adriamycin及vincristine的IC50分别为9.616 μmol/L和0.358 μmol/L,而clone #4029的IC50分别降至1.094 μmol/L和0.023 μmol/L(P<0.01),clone #4123的IC50分别降至0.780 μmol/L和0.035 μmol/L(P<0.01).COLO 320DM结肠癌亲本细胞用adriamycin及vincristine处理后其PI/AI值分别为5.68及9.59,而clone #4029的PI/AI值分别降至2.74及3.59(P<0.01),clone #4123的PI/AI值分别降至2.75及3.24(P<0.01).COLO 320DM结肠癌亲本细胞用10 μmol/L adriamycin处理后细胞内adriamycin累积浓度为27.92,而clone #4029及clone #4123细胞内adriamycin累积浓度分别增加至187.24和215.57(P<0.01).结论:稳定转染含编码MDR1 siRNA的质粒载体能稳定逆转结肠癌细胞MDR1/P-gp依赖的多药耐药性.
목적:탐토파향다약내약단백-1 (MDR1)기인적RNA간우(RNAi)대결장암세포MDR1/P-gp의뢰적다약내약성적은정역전작용.방법:분별구건함편마#4029 MDR1 siRNA화#4123 MDR1 siRNA적질립재체,병전염COLO 320DM결장암다약내약세포,채용G418사선극륭세포,경실시정량RT-PCR급Western blotting감정양성극륭세포.MTT법검측세포활력병계산각항종류약적IC50.류식세포술검측세포주기병계산PI/AI치.류식세포의측정세포내adriamycin약물루적농도.결과:양성극륭세포(clone #4029화clone #4123)적MDR1 mRNA화P-gp적표체균피억제.COLO 320DM결장암친본세포adriamycin급vincristine적IC50분별위9.616 μmol/L화0.358 μmol/L,이clone #4029적IC50분별강지1.094 μmol/L화0.023 μmol/L(P<0.01),clone #4123적IC50분별강지0.780 μmol/L화0.035 μmol/L(P<0.01).COLO 320DM결장암친본세포용adriamycin급vincristine처리후기PI/AI치분별위5.68급9.59,이clone #4029적PI/AI치분별강지2.74급3.59(P<0.01),clone #4123적PI/AI치분별강지2.75급3.24(P<0.01).COLO 320DM결장암친본세포용10 μmol/L adriamycin처리후세포내adriamycin루적농도위27.92,이clone #4029급clone #4123세포내adriamycin루적농도분별증가지187.24화215.57(P<0.01).결론:은정전염함편마MDR1 siRNA적질립재체능은정역전결장암세포MDR1/P-gp의뢰적다약내약성.