MMP-2反义寡核苷酸抑制人膀胱癌裸鼠异体移植瘤生长的实验研究

Study of MMP-2 Antisense Oligodeoxynucleotides Inhibit the Growth of Allograft Model of Human Bladder Cancer in Nude Mice

目的:探讨基质金属蛋白酶2(MMP-2)反义寡核苷酸对人膀胱癌裸鼠异体移植瘤生长的抑制作用。方法:制备膀胱癌裸鼠移植瘤模型18只,随机分为3组:MMP-2反义寡核苷酸(ASODN)治疗组、MMP-2正义寡核苷酸(SODN)治疗组及对照组,每组6只。待瘤结节直径≥5 mm后,分别在肿瘤细胞接种部位周围及中心皮下注射反义寡核苷酸/阳离子脂质体复合物、正义寡核苷酸/阳离子脂质体复合物和生理盐水。每周2次,连续4周,断颈处死裸鼠,计算肿瘤体积,称量瘤重。常规切片,HE染色,观察组织学形态并进行病理学评估。应用免疫组织化学技术(SP法)检测各组移植瘤组织中PCNA蛋白表达。结果:与对照组瘤重(7.49±0.53)g比较,ASODN组瘤重(4.18±0.53)g明显降低(P<0.01);ASODN组、正义寡核苷酸(SODN)组抑瘤率分别为44.19%、8.41%(P<0.01);ASODN组、对照组增殖指数(P1值)分别为27.63%、88.39%,抑制率为60.76%(P<0.01);ASODN组瘤体病理学特征改善。结论:MMP-2反义寡核苷酸能抑制裸鼠体内移植瘤生长,通过反义寡核苷酸下调MMP-2的表达,降低癌细胞的增殖活性,有效逆转肿瘤的恶性表型,为膀胱癌的基因治疗提供了实验依据。

Objective：To investigate the inhibitory effect of MMP- 2 antisense oligodeoxynucleotides （ASODN） on the growth of allograft model of human bladder cancer in nude mice. Methods： Mouse transplanted tumor model was established. Eighteen nude mice whose diameters of tumor nodule were more than 5mm were divided into ASODN group, sense oligodeoxynucleotides （SODN） group and control group at random. There were six mice in each group. MMP- 2 ASODN, SODN （each was mixed with Lipofectamine） and 0.9% sodium chloride solution were injected into their inoculation part of tumor cell and its central subcutaneous layer. Different treatment were given respectively at 2 times each week, continued for 4 weeks, then executed nude mice by broken the neck. The weight and volume of subcutaneous tumors were measured. HE staining, the expression of proliferating cell nuclear antigen （PCNA） was detected using immunohistochemistrical method. The pathological evaluation of tumor tissues was conducted. Results：Tumor mass in MMP 2 ASODN group decreased to 4. 18±0.53 g comparing with the control group 7.49±0.53g （P〈0.01）. The inhibition rate of tumor growth of the MMP- 2 ASODN group and MMP- 2 SODN group was 44.19% and 8.41%, respectively （ P 〈0.01）. The proliferation index of ASODN and control group was 27.63% and 88.39%, respectively. Inhibitory rate was 60.76%（ P 〈0.01）. The patho logical features of tumor tissues were ameliorated. Conclusions： MMP- 2 antisense oligodeoxynueleotides significantly decreased the growth of implanted tumor in nude mice. By ASODN reduced the expression of MMP - 2, to reduce proliferation of cancer cells, reversed effectively the malignant phenotype, and to provid an experimental basis of gene therapy for bladder cancer.