shRNA沉默IDO基因的表达对裸鼠卵巢癌腹水瘤生长的影响  

Effect of silencing IDO gene expression using shRNA targeting IDO vector on tumor growth of intraperitoneal human ovarian cancer inoculation in nude mice

作　　者：王冬冬[1] 王晓黎[2] 霍乃晨[1] 陈英汉[1] 张淑兰[1] 

机构地区：[1]中国医科大学附属盛京医院妇产科,辽宁沈阳110004 [2]中国医科大学附属第一医院内分泌科,辽宁沈阳110001

出　　处：《现代肿瘤医学》2013年第4期689-691,共3页Journal of Modern Oncology

基　　金：辽宁省博士科研启动基金资助项目(编号:20111108)

摘　　要：目的:IDO(indoleamine2,3-dioxygenase,IDO)是色氨酸代谢途径中的催化酶,其与肿瘤免疫抑制关系密切。本研究通过shRNA沉默IDO基因表达,研究抑制IDO表达在裸鼠体内对人卵巢癌移植瘤的治疗作用。方法:shRNA沉默IDO基因表达质粒和空白质粒分别稳定转染至人卵巢癌细胞SKOV-3,应用Westernblot检测IDO在SKOV-3,SKOV-3/Mock和SKOV-3/shIDO细胞中的表达情况,用MTT试剂盒检测3组肿瘤细胞体外生长速度。种植3组肿瘤细胞于裸鼠腹腔内,建立裸鼠卵巢癌腹水瘤模型,用Kaplan-Meier方法比较SKOV-3/Mock和SKOV-3/shIDO细胞接种所形成的腹水瘤裸鼠的累积生存率的差异。结果:IDO蛋白在SKOV-3/shIDO细胞中表达被抑制,在SKOV-3和SKOV-3/Mock细胞中有表达。3组肿瘤细胞体外生长曲线无统计学差异(P>0.05)。腹腔内接种SKOV-3/Mock细胞的裸鼠,均产生腹腔内转移伴腹水形成,于50天内相继死亡,而接种SKOV-3/shIDO细胞的裸鼠生存期均大于77天,此差异有显著统计学意义(P<0.01)。结论:本研究应用shRNA沉默IDO基因表达质粒稳定转染卵巢癌细胞SKOV-3,获得IDO无表达卵巢癌细胞SKOV-3/shIDO,抑制IDO表达对卵巢癌细胞体外生长速度无影响,但能抑制卵巢癌细胞腹腔内转移和腹水形成,提高累积生存率,改善预后。因此,IDO可以作为卵巢癌基因治疗的新靶点,shRNA沉默IDO基因表达可以作为卵巢癌治疗的新方法。Objective： Indoleamine 2,3 - dioxygenase （IDO） is a typtophan - catalyzing enzyme which induces immunotolerance within the tumor microenvironment. The purpose of this study is to investigate the effect of silencing IDO gene expression using shRNA targeting IDO vector on the survival curves of intraperitoneal human ovarian cancer inoculation in nude mice. Methods： SKOV -3 cells were stable transfected with shIDO expression vector and Mock vector, respectively. Western blot analysis was used to detect IDO expression in SKOV -3, SKOV -3/Mock and SK- OV - 3/shIDO cells. Cell growth curves were performed using MTT assay. Survival curves were performed using the Kaplan- Meier method after SKOV -3/Mock and SKOV -3/shIDO cells were injected intraperitoneally into nude mice, and the mice were observed until death. Results： IDO expression can be detected in the control （SKOV -3 and SKOV -3/Mock） cells but not in the SKO~ -3/shIDO cells. There was no intergroup difference in doubling time in vitro（P 〉 0.05）. All SKOV - 3/Mock cell - inoculated mice died of peritoneal dissemination with ascites within 50 days after inoculation, in contrast all SKOV - 3/shIDO cell - inoculated mice survived longer than 7 7 days （P 〈 0.01 ）. Conclusion： IDO expression can be down -regulated by shIDO vector. IDO expression did not influence the cell growth of SKOV - 3 cells in vitro. Down - regulation of IDO can improve the cumulative survival rate of ovarian cancer cell intraperitoneally inoculated nude mice. Therefore, IDO can act as a molecular therapeutic gene target of o- varian cancer and shRNA targeting IDO expression has the potential to be used as a novel therapeutic strategy against ovarian cancer.

关 键 词：IDO 卵巢癌 基因治疗 累计生存率 

分 类 号：R737.31[医药卫生—肿瘤] R737.31[医药卫生—临床医学]