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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 investigate the effect of RNA interference (RNAi) targeting hypoxia-inducible factor 1 (HIF-1) α on the growth and metastasis of colon cancer in nude mice. Methods Thirty female 6-week-old nude mice were subcutaneously implanted with colon cancer cell line SW480. The animal model of human tumor was constructed and randomly divided into normal saline group, negative plasmid group and interference group, with 10 in each group. The normal saline, negative plasmids and plasmids were injected respectively, and the growth curves were drawn. The changes of tumor growth and lymph node metastasis were observed. The changes of HIF-1α gene and protein expression were detected by RT-PCR and Western blot. Results The tumor volume of the interfering group was (273.9 ± 14.7) mm3 15 days after the transfection plasmid was introduced into the nude mice subcutaneously. Compared with the normal saline group (1845.5 ± 91.2) mm3 and the negative plasmid group (1842.7 ± 115.7) mm3, the volume was significantly (P <0.05). The rate of lymph node metastasis in the intervention group was 40% (4/10), which was significantly lower than that in the saline group (100%, 10/10) and the negative plasmid group (100%, 10/10) (χ2 = 5.952, P = 0.015). The positive rate of lymph node in the interference group was 35.7% (5/14), which was significantly lower than that in NS group (77.3%, 17/22) (χ2 = 6.420, P = 0.040). The expression of HIF-1αmRNA and HIF-1αprotein were significantly down-regulated in the interference group (P <0.05). Conclusions RNAi targeting HIF-1α can inhibit the growth and metastasis of colon cancer in nude mice and the expression of HIF-1α gene in colon cancer tissues of nude mice, which may be a target of tumor gene therapy.