摘要
目的:探讨MDR1/MRP反义寡脱氧核苷酸(Antisense oligodooxynucleotide,ASODN)片段逆转人胃腺癌耐药细胞SGC7901/ADM多药耐药(Multidrugresistance,MDR)的作用。方法:经MDR1或多药耐药相关蛋白(Multidrugresistance-asscociated protein,MRP)ASODN分别或联合转染SGC7901/ADM细胞后,采用RT-PCR法检测MDR1mRNA和MRPmRNA的表达,免疫细胞化学检测SGC7901/ADM细胞内p-糖蛋白(P-glycoprotein,P-gp)、MRP的表达,流式细胞术检测瘤细胞内Rh123的潴留状况,MTT法检测瘤细胞对阿霉素、卡铂等抗癌药的敏感性。结果:SGC7901/ADM细胞经ASODN转染12h后,MDR1和MRPmRNA的表达均降低,转染24h后下降最明显,48h后恢复至转染前水平。经转染ASODN48h后,SGC7901/ADM细胞P-gp、MRP的表达显著下降,瘤细胞内Rh123的潴留量显著高于转染前,且联合转染MDR1和MRPASODN后瘤细胞内Rh123的潴留量显著高于单纯转染组。SGC7901/ADM细胞在联合转染MDR1和MRPASODN后对阿霉素、卡铂等化疗药物的敏感性明显高于单纯转染ASODN。结论:分别转染MDR1或MRPASODN可部分逆转人胃腺癌耐药细胞SGC7901/ADM多药耐药,而联合转染2种ASODN则可显著提高瘤细胞耐药逆转效率。
Objective：To investigate the reversal effect of MDR1 and MRP antisense oligodooxynucleotide （ASODN） on adriamycin-resistant SGC7901/ADM cells. Methods：SGC7901/ADM cells were transfected by MDR1 or MRP ASODNs or the combination of two. Then the mRNA expressions of proteins MDR1 and MRP were checked by RT-PCR,and the expressions of P-gp and MRP by immunocytochemistry;the intracellular fluorescence intensity of Rhodamine 123 in these cells were determined by flow cytometry;and the sensitivity of these cells to adriamycin,carboplatin and other anticancer drugs were determined by MTr assay. Results：The expression of MDRlmRNA and MRP mRNA in SGC7901/ADM cells decreased at 12h after transfection of ASODNs,decreased to the lowest level at 24h and returned to the level before transfection after 48h. The expression of P-gp and MRP decreased significantly at 48h after transfection with ASODNs compared with the control ceils. The retention of Rhodamine 123 in SGC7901/ADM ceils was significantly higher than that before transfection, and the intracellular fluorescence intensity significantly increased in ASODNs cotransfection SGC7901/ADM cells, compared with those transfected by MDR1 or MRP ASODNs respectively;In addition,the sensitivity of SGC7901/ADM cells to adriamycin, carboplatin and other anticancer drugs obviously increased after cotransfection of MDR1 and MRP ASODNs compared with transfection of MDR1 or MRP ASODN respectively. Conclusion：The transfection of MDR1 or MRP ASODN can partly reverse the muhidrug resistance of gastric glandular carcinoma ceils SGC7901/ADM,while cotransfection of MDR1 and MRP ASODNs can significantly reverse drug-resistance of these tumor cells.
出处
《重庆医科大学学报》
CAS
CSCD
2008年第6期686-690,共5页
Journal of Chongqing Medical University
关键词
反义寡核苷酸
胃癌细胞
逆转效率
多药耐药
Antisense oligodooxynucleotide（ASODN ）
Gastric carcinoma cells
Reversal effect
Multidrug resistance
