CAJ | 학술논문

目的：探讨miRNA-181a对乳腺癌耐药蛋白( BCRP)及其介导的乳腺癌细胞耐药性的调控作用。方法应用生物信息学预测 BCRP mRNA-3ˊUTR 与 miR-181a 的结合位点；荧光素酶报告分析检测miR-181a与BCRP mRNA-3ˊUTR的结合作用；qRT-PCR和Western blot检测细胞中相关蛋白表达水平。结果与阴性转染组比较,miR-181a mimic 与PGL3-BCRP 3ˊUTR共转染后,荧光素酶活性明显降低( P<0.05)。 miR-181a mimic转染到MCF7/MX细胞48 h后,与阴性转染组相比,导致原本 miR-181a低表达的 MCF-7/MX细胞中miR-181a的表达增加(P<0.05),BCRP mRNA和蛋白表达明显下降(P<0.05),而MRP、P-gp、LRP的mRNA和蛋白水平均无明显改变(P>0.05)；MCF-7细胞在miR-181a inhibitor转染48 h后,与阴性转染组相比,miR-181a的表达降低( P<0.05)。同时, BCRP mRNA和蛋白表达明显增加(P<0.05)。而MRP、P-gp、LRP的mRNA和蛋白水平无明显改变(P>0.05)。结论 miR-181a可通过靶向作用于BCRP mRNA-3ˊUTR区,调控BCRP表达。
목적：탐토miRNA-181a대유선암내약단백( BCRP)급기개도적유선암세포내약성적조공작용。방법응용생물신식학예측 BCRP mRNA-3ˊUTR 여 miR-181a 적결합위점；형광소매보고분석검측miR-181a여BCRP mRNA-3ˊUTR적결합작용；qRT-PCR화Western blot검측세포중상관단백표체수평。결과여음성전염조비교,miR-181a mimic 여PGL3-BCRP 3ˊUTR공전염후,형광소매활성명현강저( P<0.05)。 miR-181a mimic전염도MCF7/MX세포48 h후,여음성전염조상비,도치원본 miR-181a저표체적 MCF-7/MX세포중miR-181a적표체증가(P<0.05),BCRP mRNA화단백표체명현하강(P<0.05),이MRP、P-gp、LRP적mRNA화단백수평균무명현개변(P>0.05)；MCF-7세포재miR-181a inhibitor전염48 h후,여음성전염조상비,miR-181a적표체강저( P<0.05)。동시, BCRP mRNA화단백표체명현증가(P<0.05)。이MRP、P-gp、LRP적mRNA화단백수평무명현개변(P>0.05)。결론 miR-181a가통과파향작용우BCRP mRNA-3ˊUTR구,조공BCRP표체。
Aim ToinvestigatetheeffectsofmiRNA-181 a on breast cancer resistance protein ( BCRP ) . Methods Bioinformaticspredictedbindingsitesof BCRP mRNA-3ˊUTR region and miR-181 a;further lu-ciferase reporter gene analysis confirmed that miR-181 a could combine with BCRP mRNA-3ˊUTR; qRT-PCR and Western blot detected related mRNA and protein expressionlevels.Results Comparedwithnegative transfection group, after the miR-181a mimic and PGL3-BCRP 3ˊUTR were co-transfected, luciferase ac-tivity was significantly decreased ( P <0 . 05 ) . After miR-181a mimic transfected MCF7/MX cells for 48h, compared to the negative group, the expression of miR-181 a in MCF-7/MX cells was increased ( P<0. 05 ) , BCRP mRNA, BCRP protein expression were signifi-cantly decreased ( P<0 . 05 ) , while mRNA expression and protein levels of MRP, P-gp, LRP did not change significantly ( P >0. 05 ); after transfecting miR-181 a inhibitor for 48h, compared to the negative group, the expression of miR-181 a in MCF-7 cells was reduced (P<0. 05). Meanwhile,BCRP mRNA expression and BCRP protein expression were also significantly in-creased ( P<0. 05 ) . The mRNA expression and pro-tein levels of MRP, P-gp, LRP did not change signifi-cantly(P>0.05).Conclusion miR-181acanregu-late BCRP expression by targeting the BCRP mRNA-3ˊUTR region.