CAJ | 학술논문

目的：探讨利用载小干扰RNA（small interfering RNA, siRNA）的脂质微泡超声造影剂实现恶性肿瘤基因治疗与疗效评估一体化的可行性。方法以异质性组装法制备载siRNA的脂质微泡，并用动态光散射法检测其直径大小和表面电位。通过激光共聚焦显微镜观察红色荧光标记的siRNA在瘤内的分布。针对抗凋亡基因sirtuin 2（SIRT2）设计siRNA，通过在体实验研究载siRNA微泡在超声辐照下的肿瘤基因沉默效果。在用载siRNA微泡治疗肿瘤的同时，以超声造影技术观察肿瘤治疗效果。结果 siRNA微泡的直径为400.7±30.5 nm，表面带弱正电（+8.8±0.8 mV）。siRNA微泡协同超声辐照能高效地将siRNA递送到肿瘤细胞胞浆内，有效沉默肿瘤组织中的SIRT2基因，诱导肿瘤凋亡，明显减缓肿瘤的生长速度。超声造影检查结果提示，载siRNA微泡具有良好的超声显像效果，能在治疗过程中实时评估肿瘤的血供情况。结论新型载siRNA的脂质微泡超声造影剂在活体上能对肿瘤进行基因沉默治疗，同时观察肿瘤治疗效果，实现恶性肿瘤基因治疗与疗效评估的一体化。
목적：탐토이용재소간우RNA（small interfering RNA, siRNA）적지질미포초성조영제실현악성종류기인치료여료효평고일체화적가행성。방법이이질성조장법제비재siRNA적지질미포，병용동태광산사법검측기직경대소화표면전위。통과격광공취초현미경관찰홍색형광표기적siRNA재류내적분포。침대항조망기인sirtuin 2（SIRT2）설계siRNA，통과재체실험연구재siRNA미포재초성복조하적종류기인침묵효과。재용재siRNA미포치료종류적동시，이초성조영기술관찰종류치료효과。결과 siRNA미포적직경위400.7±30.5 nm，표면대약정전（+8.8±0.8 mV）。siRNA미포협동초성복조능고효지장siRNA체송도종류세포포장내，유효침묵종류조직중적SIRT2기인，유도종류조망，명현감완종류적생장속도。초성조영검사결과제시，재siRNA미포구유량호적초성현상효과，능재치료과정중실시평고종류적혈공정황。결론신형재siRNA적지질미포초성조영제재활체상능대종류진행기인침묵치료，동시관찰종류치료효과，실현악성종류기인치료여료효평고적일체화。
Objective To evaluate the feasibility of integrating cancer gene therapy with therapeutic effect evaluation using siRNA-loaded nano-scale microbubbles (siRNA-NBs). Methods siRNA-NBs were prepared by hetero-assembly of polymeric siRNA micelles and liposomal microbubbles, and the particle sizes and surface potentials were examined with dynamic light scattering. The distributions of cy3- labled siRNA in the tumor tissues were evaluated using confocal laser scanning microscopy. A siRNA targeting the anti-apoptosis gene SIRT2 was designed and its gene silencing effects was tested in vivo using siRNA-NBs with ultrasound exposure. The therapeutic effect of the loaded siRNA-NBs was evaluated by contrast-enhanced ultrasonography. Results The siRNA-NBs had a mean diameter of 400.7 ± 30.5 nm with a weak positive charge of+8.8 ± 0.8 mV. With ultrasound exposure, siRNA-NBs effectively delivered cy3-siRNA into the cytoplasm of cancer cells and caused SIRT2 suppression and cell apoptosis in tumor tissues, resulting in significantly suppressed tumor growth. In addition, contrast-enhanced ultrasonography of siRNA-NBs provided good imaging quality to allow real-time observation of blood supply during gene therapy. Conclusions As a novel ultrasound contrast agent, siRNA-NBs make possible the integration of tumor gene therapy and therapeutic effect evaluation for cancer.