CAJ | 학술논문

本研究构建了能够靶向肿瘤新生血管的RGD修饰阳离子脂质体(RGD-Lipo),作为靶向耐药相关基因MDR1的siRNA输送载体并评价其相关的药剂学性质.该脂质体与siRNA形成的复合物粒径控制在200 nm以内,并且对其中所包载的siRNA具有一定的保护作用.体外实验结果表明,经RGD修饰的脂质体(RGD-Lipo)细胞粘附能力显著增强,并可增加细胞内siRNA的转染效果.与利用前插法进行靶向修饰相比,利用后插法进行RGD修饰可有效地改善siRNA的溶酶体释放效率.细胞毒试验结果表明,后插法制备的pRGD-Lipo-siRNA能够有效逆转人卵巢癌SKVO3/A细胞的耐药性,并增加阿霉素药物在细胞内的蓄积.体外研究结果证实,使用pRGD-Lipo-siRNA有利于提高耐药细胞对化疗药阿霉素的敏感度,并将有可能应用于临床耐药肿瘤的治疗.
본연구구건료능구파향종류신생혈관적RGD수식양리자지질체(RGD-Lipo),작위파향내약상관기인MDR1적siRNA수송재체병평개기상관적약제학성질.해지질체여siRNA형성적복합물립경공제재200 nm이내,병차대기중소포재적siRNA구유일정적보호작용.체외실험결과표명,경RGD수식적지질체(RGD-Lipo)세포점부능력현저증강,병가증가세포내siRNA적전염효과.여이용전삽법진행파향수식상비,이용후삽법진행RGD수식가유효지개선siRNA적용매체석방효솔.세포독시험결과표명,후삽법제비적pRGD-Lipo-siRNA능구유효역전인란소암SKVO3/A세포적내약성,병증가아매소약물재세포내적축적.체외연구결과증실,사용pRGD-Lipo-siRNA유리우제고내약세포대화료약아매소적민감도,병장유가능응용우림상내약종류적치료.
In this study, the novel RGD-modified stabilized cationic liposomes were developed as the delivery vehicle for siRNA targeting human MDR1 gene. The complex of cationic liposomes and siRNA, RGD-Lipo-siRNA, was prepared with a narrow size distribution below 200 nm. It was shown that the encapsulated siRNA in the liposomes could be effectively protected from serum degradation. Also, enhanced cell binding and intracellular uptake of siRNA in the doxorubicin-resistant human ova- rian cancer cell lines SKOV3/A were found in RGD-Lipo-siRNA preparation as compared to that of unmodified cationic lipsomes (Lipo-siRNA). Using the post-insertion method for RGD modification, lysosome release of siRNA in pRGD-Lipo-siRNA was improved. From flow cytometry, significant increase of doxorubicin accumulation was observed in the SKOV3/A cells treated with pRGD-Lipo-siRNA targeting human MDR1 gene. In vitro eytotoxicity assay showed that the significant cell growth inhibi- tion was achieved in the SKOV3/A cells after treating with the combined use of siRNA and doxorubicin. In conclusions, post- inserted RGD modified lipoplex, pRGD-Lipo-siRNA, was successfully used for siRNA transfection and achieved drug resistance reversal in human ovarian cancer SKOV3/A (doxorubicin-resistant) cells. It suggested that this liposomes might be a potential vehicle for siRNA delivery in vivo.