siRNA干扰血小板内皮细胞黏附因子-1表达对人鼻咽癌耐药细胞CNE1/R多药耐药性的影响  

Effect of siRNA interference with PECAM-1 expression on multidrug resistance of human nasopharyngeal carcinoma drug-resistant cells CNE1/R

作　　者：蒋中秀[1] 李昕[1] 伦永志[2] 王若雨[1] 

机构地区：[1]大连大学附属中山医院肿瘤科,辽宁省116001 [2]大连大学医学院辽宁省高校生物物理学重点实验室

出　　处：《中华临床医师杂志（电子版）》2012年第10期I0001-I0005,共5页Chinese Journal of Clinicians(Electronic Edition)

基　　金：国家自然科学基金(30970869)

摘　　要：目的探讨抑制血小板内皮细胞黏附因子-1(PECAM-1)表达对多药耐药基因1(MDR-1)及其编码产物P-糖蛋白(P-gp)的影响。方法设计并化学合成针对PECAM-1的三条siRNA序列,命名为si-PECAM1,2,3,脂质体转染法将其转入CNE1/R,采用Semi-qRT-PCR、Westernblot方法检测并筛选出抑制效果最好的si-PECAM序列,将筛选出的si-PECAM转入CNE1/R,采用Semi-qRT-PCR、Western blot方法检测抑制PECAM-1表达前后MDR-1及其编码产物P-gp的变化情况。结果 Semi-qRT-PCR的结果显示,设计三条si-PECAM序列均有效,以si-PECAM3干扰效果最明显(P<0.001),Western blot结果显示与Semi-qRT-PCR的结果一致;转染si-PECAM348h后做Semi-qRT-PCR的结果显示抑制PECAM-1表达后MDR-1表达明显下调(P<0.001),Western blot结果与Semi-qRT-PCR的结果一致,抑制PECAM-1表达后MDR-1编码产物P-gp的表达也明显下调(P<0.001)。结论抑制CNE1/R中PECAM-1表达后MDR-1及其编码产物P-gp的表达均明显下调,提示PECAM-1可能参与X射线辐射诱导人鼻咽癌细胞发生多药耐药。Objective To inhibit the expression of PECAM-1 with the technology of siRNA in human nasopharyngeal carcinoma drug-resistant cells CNE1/R, induced by X-ray radiation, then detect the expression of MDR-1 and its encoded product P -gp with the methods of Semi -qRT-PCR and Western blot, and discuss the effects of PECAM-I on MDR-1 and its encoded product P-gp. Methods 3 siRNA sequences were designed and chemical synthesis for PECAM-1, named si-PECAM1,2, 3, transferred into CNE1/R. Detecting and filtering out the best inhibition effeet＇s si-PECAM sequence with Semi-qRT-PCR and Western blot, then transferred into CNEI/R, detecting the changes of MDR-1 and its encoded product P-gp with Semi-qRT-PCR and Western blot before and after inhibiting the expression of PECAM-1. Results Semi-qRT-PCR results showed that 3 si-PECAM sequences were effective,the most significant interference effect was si-PECAM3 （P 〈 0. 001 ）, and Western blot results were consistent with Semi-qRT-PCR results. After transfected si-PECAM3 for 48 h, Semi-qRT-PCR results showed that the expression of MDR-1 was significantly decreased （P 〈 0. 001 ）, Western blot results were consistent with Semi -qRT- PCR results, the expression of P-gp was significantly decreased （ P 〈 0. 001 ）. Conclusions After inhibiting the expression of PECAM-1 ,the expression of MDR-1 and P-gp were significantly decreased, suggesting that PECAM-1 may be involved in X-ray radiation induced the occurrence of multidrug resistance in nasopharyngeal carcinoma cells .

关 键 词：鼻咽肿瘤 RNA 小分子干扰 抗原 CD31 抗药性 多药 

分 类 号：R739.63[医药卫生—肿瘤]