白血病·淋巴瘤
白血病·淋巴瘤
백혈병·림파류
JOURNAL OF LEUKEMIA & LYMPHOMA
2010年
5期
276-280
,共5页
张丽%卓家才%张琼丽%陶小梅%楼瑾%史敦云%李明%杜新
張麗%卓傢纔%張瓊麗%陶小梅%樓瑾%史敦雲%李明%杜新
장려%탁가재%장경려%도소매%루근%사돈운%리명%두신
肿瘤,实验性%腺病毒载体%RNA%砷剂%抗药性,肿瘤%基因,MRP1%K562细胞
腫瘤,實驗性%腺病毒載體%RNA%砷劑%抗藥性,腫瘤%基因,MRP1%K562細胞
종류,실험성%선병독재체%RNA%신제%항약성,종류%기인,MRP1%K562세포
Neoplasms,experimental%Adenovirus vector%RNA%Arsenicals%Drug resistance,neoplasm%Genes,MRP1%K562 cells
目的 构建MRP1特异性发夹状RNA(shRNA)重组腺病毒载体,并研究其对耐三氧化二砷(ATO)的白血病K562/AS2细胞MRP1基因表达的抑制作用.方法 构建MRP1特异性shRNA重组腺病毒,并感染K562/AS2细胞;用荧光实时定量PCR法分析MRP1 mRNA的表达水平;流式细胞术检测MRP1蛋白表达;MTT法检测该细胞对ATO及依托泊苷(VP_(16))的细胞毒作用.结果 pAd-EGFP-U6.MRP1.shRNA重组腺病毒感染前后,K562/AS2细胞中MRP1 mRNA和蛋白表达水平分别为(34.70±0.28)、(4.19±0.03)(P<0.05)和(26.40±0.16)、(10.85±0.37)(P<0.05);感染前后K562/AS2细胞对ATO及VP16耐药倍数分别为(11.4078±0.3183)、(1.6126±0.3015)和(5.9141±0.0149)、(1.7664±0.1038),逆转倍数分别为(7.2409±1.3668)和(3.3555±0.1886)(P<0.05).结论 成功构建了pAd-EGFP-U6-MRP1.shRNA重组腺病毒载体,该载体感染K562/AS2细胞后可抑制细胞MRP1基因表达以及逆转该细胞对ATO和VP16的耐药.
目的 構建MRP1特異性髮夾狀RNA(shRNA)重組腺病毒載體,併研究其對耐三氧化二砷(ATO)的白血病K562/AS2細胞MRP1基因錶達的抑製作用.方法 構建MRP1特異性shRNA重組腺病毒,併感染K562/AS2細胞;用熒光實時定量PCR法分析MRP1 mRNA的錶達水平;流式細胞術檢測MRP1蛋白錶達;MTT法檢測該細胞對ATO及依託泊苷(VP_(16))的細胞毒作用.結果 pAd-EGFP-U6.MRP1.shRNA重組腺病毒感染前後,K562/AS2細胞中MRP1 mRNA和蛋白錶達水平分彆為(34.70±0.28)、(4.19±0.03)(P<0.05)和(26.40±0.16)、(10.85±0.37)(P<0.05);感染前後K562/AS2細胞對ATO及VP16耐藥倍數分彆為(11.4078±0.3183)、(1.6126±0.3015)和(5.9141±0.0149)、(1.7664±0.1038),逆轉倍數分彆為(7.2409±1.3668)和(3.3555±0.1886)(P<0.05).結論 成功構建瞭pAd-EGFP-U6-MRP1.shRNA重組腺病毒載體,該載體感染K562/AS2細胞後可抑製細胞MRP1基因錶達以及逆轉該細胞對ATO和VP16的耐藥.
목적 구건MRP1특이성발협상RNA(shRNA)중조선병독재체,병연구기대내삼양화이신(ATO)적백혈병K562/AS2세포MRP1기인표체적억제작용.방법 구건MRP1특이성shRNA중조선병독,병감염K562/AS2세포;용형광실시정량PCR법분석MRP1 mRNA적표체수평;류식세포술검측MRP1단백표체;MTT법검측해세포대ATO급의탁박감(VP_(16))적세포독작용.결과 pAd-EGFP-U6.MRP1.shRNA중조선병독감염전후,K562/AS2세포중MRP1 mRNA화단백표체수평분별위(34.70±0.28)、(4.19±0.03)(P<0.05)화(26.40±0.16)、(10.85±0.37)(P<0.05);감염전후K562/AS2세포대ATO급VP16내약배수분별위(11.4078±0.3183)、(1.6126±0.3015)화(5.9141±0.0149)、(1.7664±0.1038),역전배수분별위(7.2409±1.3668)화(3.3555±0.1886)(P<0.05).결론 성공구건료pAd-EGFP-U6-MRP1.shRNA중조선병독재체,해재체감염K562/AS2세포후가억제세포MRP1기인표체이급역전해세포대ATO화VP16적내약.
Objective To construct a recombinant adenovirus vector of Hairpin RNA specific for MRP1 gene and study its inhibition of MRP1 gene expression in K562/AS2 cell line resistant to AS_2O_3 (ATO). Methods A MRP1-specific hairpin RNA recombinant adenovirus vector was constructed and used to infected K562/AS2 cells. Expression level of MRP1 mRNA detected by real-time fluorescent quantitative PCR. MRP1 protein detected by flow cytometry. MTT method was used to detected the cytotoxicity of ATO and etoposide. Results MRP1 mRNA and protein expression level in K562/AS2 cells before and after the pAd-MRPl-shRNA adenovirus infection was (34.70±0.28 vs 4.19±0.03, P <0.05) and (26.40±0.16 vs 10.85±0.37, P<0.05), respectively. RR of K562/AS2 to arsenic trioxide and etoposide was (11.4078±0.3183 fold vs 1.6126±0.3015 fold, P<0.05) and (5.9141 ±0.0149 fold vs 1.7664±0.1038 fold, P <0.05), respectively. The reversal fold of ATO and etoposide was (7.2409±1.3668) and (3.3555±0.1886), respectively. Conclusion Successfully constructed pAd-EGFP-U6-shRNA-MRPl adenovirus vector, the vector of infection K562/SA2 cells can inhibit MRP1 gene expression and reverse the resistance of the ATO and etoposide.
