Department of Nuclear Medicine, School of Medicine, Kyungpook National University, Daegu, Republic of Korea.
Mol Imaging. 2010 Dec;9(6):343-50.
The reversal effect of multidrug resistance (MDR1) gene expression by adenoviral vector-mediated MDR1 ribonucleic acid interference was assessed in a human colon cancer animal model using bioluminescent imaging with Renilla luciferase (Rluc) gene and coelenterazine, a substrate for Rluc or MDR1 gene expression. A fluorescent microscopic examination demonstrated an increased green fluorescent protein signal in Ad-shMDR1- (recombinant adenovirus that coexpressed MDR1 small hairpin ribonucleic acid [shRNA] and green fluorescent protein) infected HCT-15/Rluc cells in a virus dose-dependent manner. Concurrently, with an increasing administered virus dose (0, 15, 30, 60, and 120 multiplicity of infection), Rluc activity was significantly increased in Ad-shMDR1-infected HCT-15/Rluc cells in a virus dose-dependent manner. In vivo bioluminescent imaging showed about 7.5-fold higher signal intensity in Ad-shMDR1-infected tumors than in control tumors (p < .05). Immunohistologic analysis demonstrated marked reduction of P-glycoprotein expression in infected tumor but not in control tumor. In conclusion, the reversal of MDR1 gene expression by MDR1 shRNA was successfully evaluated by bioluminescence imaging with Rluc activity using an in vivo animal model with a multidrug resistance cancer xenograft.
采用带有海肾荧光素酶（RLuc）基因和腔肠素的生物发光成像，评估了腺病毒载体介导的 MDR1 核糖核酸干扰对人结肠癌动物模型中多药耐药（MDR1）基因表达的逆转作用，腔肠素是 RLuc 或 MDR1 基因表达的底物。荧光显微镜检查显示，Ad-shMDR1-（共表达 MDR1 短发夹 RNA（shRNA）和绿色荧光蛋白的重组腺病毒）感染的 HCT-15/RLuc 细胞中的绿色荧光蛋白信号呈病毒剂量依赖性增加。同时，随着给予病毒剂量的增加（0、15、30、60 和 120 感染复数），RLuc 活性在 Ad-shMDR1 感染的 HCT-15/RLuc 细胞中呈病毒剂量依赖性显著增加。体内生物发光成像显示，Ad-shMDR1 感染的肿瘤信号强度比对照肿瘤高约 7.5 倍（p <.05）。免疫组织化学分析显示，感染肿瘤中 P-糖蛋白表达明显减少，但对照肿瘤中没有。总之，通过使用多药耐药性癌症异种移植的体内动物模型，通过 RLuc 活性的生物发光成像成功评估了 MDR1 shRNA 对 MDR1 基因表达的逆转作用。
