摘要
目的探讨靶向缺氧诱导因子-1(hypoxia-inducible factor1,HIF-1)α的RNA干扰(RNA interference,RNAi)对裸鼠结肠癌生长及转移的影响。方法利用结肠癌细胞株SW480对30只6周龄雌性裸鼠进行皮下种植,构建荷人瘤动物模型后按随机数字表法分为生理盐水组、阴性质粒组及干扰组,每组10只。分别注射生理盐水、阴性质粒及转染质粒,并绘制肿瘤生长曲线,观察各组间肿瘤生长及淋巴结转移的变化,采用RT-PCR和Western blot检测HIF-1α基因及蛋白表达的变化。结果转染质粒导入裸鼠皮下移植瘤后15d,干扰组肿瘤体积为(273.9±14.7)mm3,与生理盐水组(1845.5±91.2)mm3和阴性质粒组(1842.7±115.7)mm3相比较,体积明显减小(P<0.05);干扰组淋巴结转移率为40%(4/10),与生理盐水组(100%,10/10)及阴性质粒组(100%,10/10)相比明显降低(χ2=5.952,P=0.015);干扰组淋巴结阳性率为35.7%(5/14),与生理盐水组77.3%(17/22)及阴性质粒组65.2%(15/23)相比明显降低(χ2=6.420,P=0.040)。干扰组肿瘤组织中HIF-1αmRNA和HIF-1α蛋白显著下调(P<0.05)。结论靶向HIF-1α的RNAi可以抑制裸鼠结肠癌的生长及淋巴结转移和裸鼠结肠癌组织内HIF-1α基因的表达,可能成为肿瘤基因治疗的一个靶点。
Objective To explore the influence of RNAi-mediated hypoxia-inducible factor 1α （HIF-1α） silencing on the growth and metastasis of colon cancer in nude mice.Methods A total of 30 6-week-old female nude mice were randomly divided into normal saline group （NS group）,negative plasmid group and RNA interference group after injected with SW480 subcutaneously to form a colon carcinoma.shRNA-HIF-1α vector was constructed and injected （50 μg/time,once per 2 d,for 7 times） into the tumor mass of nude mice of RNA interference group to induce RNAi.Normal saline or blank vector was injected into the corresponding nude mice.The changes of HIF-1α were detected with RT-PCR and Western blot analysis.The size of tumors,metastasis rate and the positive rate of lymphnode were compared among different groups.Results In 15 d after RNAi vector injection,the tumor volume of RNAi group [（273.9±14.7） mm3] was much smaller than those of NS group [（1 845.5±91.2） mm3] and negative plasmid group [（1 842.7±115.7） mm3] （P0.05）.Lymph node metastasis rate in RNAi group （40%,4/10） was lower than those of NS group （100%,10/10） and negative plasmid group （100%,10/10）（Chi square=5.952,P=0.015）.The lymph node positive rate in experimental group （35.7%,5/14） was significantly lower than those of NS group （77.3%,17/22） and negative plasmid group （65.2%,15/23）（Chi square=6.420,P=0.040）.The mRNA level of HIF-1α and its protein expression of RNAi group also reduced obviously （P0.05）.Conclusion The expression of HIF-1α,tumor sizes,metastasis rate and the positive rate of lymphnode in nude mouse have been obviously inhibited by pshRNA-HIF-1α,which may be used as a target for gene therapy.
出处
《第三军医大学学报》
CAS
CSCD
北大核心
2010年第9期908-911,共4页
Journal of Third Military Medical University
基金
重庆市卫生局2009年度医学科研计划项目重点项目(2009-1-49)~~
