低强度超声介导的载双自杀基因纳泡对膀胱癌小鼠模型的治疗研究

Therapeutic effect of low intensity ultrasound-mediated nanobubbles loaded with double suicide gene on bladder cancer in mice

摘要:

目的：制备载胞苷脱氨酶-胸腺嘧啶激酶（colicytocinedeaminase-thymidinekinase，CD-TK）双自杀基因纳泡，用于治疗膀胱癌小鼠。方法：采用薄层法制备载CD-TK双自杀基因纳泡，合成CD-TK双自杀基因阳离子纳泡，光镜观察纳泡的形态，马尔文粒径仪测其粒径；建立裸鼠后膀胱癌模型，超声辐照肿瘤部位，转染双自杀基因，按实验分组每组连续腹腔注射无毒前药1周，开始记录每只裸鼠肿瘤的最长径和最短径，计算并记录肿瘤的体积变化；第5周取裸鼠肿瘤标本，免疫荧光评估肿瘤组织切片细胞凋亡和增殖情况。结果：本研究成功制备了载CD-TK双自杀基因纳泡。通过超声介导载CD-TK纳泡成功转染自杀基因，抑制肿瘤增殖和促进肿瘤凋亡。结论：载CD-TK纳泡可能成为治疗膀胱癌的一种新方式。

Abstract:

Objective：To prepare nanobubbles loaded with colicytocinedeaminase-thymidinekinase（CD-TK） for treating bladder cancer in mice. Methods：Nanobubbles loaded with CD-TK double suicide gene were prepared by thin layer method and cationic nanobub－bles loading CD-TK double suicide gene were synthesized. Nanobubbles’ shape was observed through light microscope and the par－ticle diameter was measured via Malvin laser particle size analyzer. The mice model with bladder cancer was established，ultrasound was used to irradiate the tumor site and the double suicide gene was transfected. Intraperitoneal injection of atoxic prodrug was con－tinuously given to each group for one week. The longest and shortest diameters of each nude mouse’s tumor were recorded，and the tumor’s volume changes were calculated and recorded. Tumor specimens were taken at the fifth week. Apoptosis and proliferation of tumor cells were evaluated by immunofluorescence. Results：The nanobubbles loading CD-TK double suicide gene were successfully prepared in this study. Ultrasound-mediated nanobubbles loaded with CD-TK can successfully transfect suicide gene，inhibiting the proliferation and apoptosis of tumors. Conclusion：Nanobubbles loaded with CD-TK may be a new therapy for bladder cancer.