CYP3A5基因反义寡核苷酸转染K562/A02细胞逆转耐药的研究  

Study on the reversal of the drug-resistant phenotype of K562/A02 cells transfected the antisense oligonucleotides complementary to CYP3A5 gene

作　　者：王婷[1] 陈芳源[1] 钟华[1] 黄洪辉[1] 钟济华[1] 欧阳仁荣[1] 

机构地区：[1]上海第二医科大学附属仁济医院血液科,上海血液研究所,上海200001

出　　处：《诊断学理论与实践》2004年第6期413-415,419,共4页Journal of Diagnostics Concepts & Practice

基　　金：国家自然科学基金资助项目(编号30400183)

摘　　要：目的:探讨针对细胞色素P4503A亚家族多肽5(CYP3A5)耐药机制进行反义寡核苷酸逆转耐药。方法:CYP3A5基因反义寡核苷酸转染K562/A02细胞后,RT鄄PCR方法检测反义寡核苷酸对CYP3A5基因转录的特异性阻断,MTT法测定不同处理组柔红霉素IC50值。结果:针对CYP3A5基因的反义寡核苷酸转染K562/A02细胞48h后明显抑制CYP3A5基因的转录,而正义、错义寡核苷酸处理组则无明显改变。转染针对CYP3A5基因的反义寡核苷酸,可使柔红霉素对K562/A02细胞的IC50值由(7.728±3.625)mg/L降至(2.494±1.674)mg/L,增敏3.10倍,差异有显著性(P<0.05),而正义、错义寡核苷酸处理组IC50值与对照组相比则无明显差异。结论:针对CYP3A5基因的反义寡核苷酸可显著逆转K562/A02细胞的耐药性。Objective To investigate if the CYP3A5-mediated drug resistance can be reversed by the transfection of the antisense oligonucleotides. Methods The specific inhibition to the transcription of CYP3A5 gene was detected using RT-PCR assay after the transfection of the antisense oligonucleotides to K562/A02 cells. Daunorubicin IC50 values of K562/A02 cells treated with antisense, sense and mismatch oligonucleotides were obtained by MTT assay. Results Transfecting K562/A02 cells with antisense oligonucleotides complementary to CYP3A5 gene remarkably inhibited the CYP3A5 gene transcription after 48 hours, and statistically reduced daunorubicin IC50 values of K562/A02 cells from (7.728±3.625) mg/L to (2.494±1.674) mg/L (P< 0.05). There was no significant difference between the sense and the mismatch oligonucleotides treatment groups. Conclusions Antisense oligonucleotides complementary to CYP3A5 gene may obviously reverse the drug-resistant phenotype of K562/A02 cells.

关 键 词：CYP3A5基因 反义寡核苷酸 K562/A02细胞 基因转染 耐药机制 

分 类 号：R394[医药卫生—医学遗传学]