环氧合酶-2反义核酸对膀胱癌细胞恶性表型的抑制作用  

The inhibition of malignant phenotypes of human bladder cancer cells by anti-sense nucleic acid of cyclooxygenase-2

作　　者：秦军[1] 吴开春[2] 王禾[1] 陈宝琦[1] 幺立萍[2] 邵国兴[1] 

机构地区：[1]第四军医大学西京医院泌尿外科,西安710032 [2]第四军医大学西京医院消化内科,西安710032

出　　处：《中华泌尿外科杂志》2006年第4期235-238,共4页Chinese Journal of Urology

摘　　要：目的探讨环氧合酶-2(COX-2)反义核酸对膀胱癌细胞恶性表型的抑制作用。方法采用脂质体介导方法,分别构建转染COX-2反义真核表达载体和空载体的膀胱癌细胞系5637-AS细胞和5637-P细胞,RT-PCR及W estern b lot分析转染细胞COX-2 mRNA及蛋白表达水平。MTT比色实验、Boyden侵袭小室法和裸鼠成瘤实验分别检测转染细胞体外增殖速度、体外侵袭力和体内成瘤性。结果RT-PCR结果显示,5637-AS细胞COX-2/磷酸甘油醛脱氢酶(GAPDH)mRNA比值(0.65)明显低于5637(0.92)和5637-P细胞(0.90);W estern b lot提示5637-AS细胞COX-2/β-actin比值(0.29)明显低于5637细胞(0.46)和5637-P细胞(0.44)。MTT比色实验显示5637-AS细胞较5637-P、5637细胞生长速度减慢,三者倍增时间分别为3.6 d、2.9 d和2.9 d。体外侵袭实验结果表明,5637-AS侵袭细胞数显著低于5637细胞及5637-P细胞(18.20±5.97比45.40±8.12,18.20±5.97比44.10±6.47,P<0.01)。裸鼠成瘤实验中,5637-P、5637细胞接种裸鼠后第5天肿瘤长出,而5637-AS细胞第7天肿瘤长出;30 d后5637-AS细胞接种组瘤体重量(392.15 mg±33.58 mg)明显小于5637细胞(738.26 mg±66.32 mg),和5637-P细胞接种组(698.53 mg±88.76 mg),P<0.01。结论膀胱癌细胞中COX-2过表达与细胞的恶性表型相关,反义技术抑制COX-2表达可以逆转膀胱癌细胞的恶性表型。Objective To explore the effect of anti-sense nucleic acid of cyclooxygenase-2（COX-2） in malignant phenotype of bladder cancer cells and the mechanism of COX-2 in carcinogenesis of bladder cancer. Methods Using lipofectAMINETM2000 reagent, the COX-2 highly expressed in human bladder cancer cell line 5637 was transfected with anti-sense eukaryotic expression vector pcDNA3.1/hCOX2 （ - ） and control plasmid pcDNA3.1 （named as 5637-AS and 5637-P cells, respectively）. Semi-quantitative RTPCR and Western blot were used to testify the mRNA and protein level in the transfected cells. MTT assay, Boyden chamber and tumor implantations experiment were used to detect the proliferation, invasion and tumorigenesis of the transfected cells in vitro and in vivo. Results RT-PCR analysis indicated that the ratio of COX-2 mRNA to GAPDH mRNA in 5637-AS cells was significantly lower than 5637 and 5637-P cells （0.65 vs 0.92 ;0.65 vs 0.90 ,respectively）. Western blot showed that the ratio of COX-2 protein to β-actin protein in 5637-AS （0.29） was significantly lower than in 5637 （0.46） and 5637-P （0.44）. MTT assay suggested that the 5637-AS cells proliferated more slowly than the 5637 and 5637-P cells with double time of 3.6 d ,2.9 d and 2.9 d, respectively. Boyden chamber assay revealed that the cell number of the 5637-AS cells that migrated through the filter was less than the 5637 and 5637-P cells （ 18.20 ± 5.97 vs 45.40 ±8, 12,18.20 ± 5, 97 vs 44.10 ±6. 47, respectively ;P 〈 0. 01 ）, The tumor xenografts occurred at 5 d in 5637 and 5637-P cells ,while at 7 d in 5637-AS cells. At 30 d after implantation, the weight of 5637-AS cell tumor xenografts （392.15 ± 33, 58）mg was significantly less than that of 5637 cells （738.26 ± 66, 32）mg and 5637-P ceils （698.53±88,76）mg（P〈0.01）. Conclusions Overexpression of COX-2 in human bladder cancer cell line 5637 is associated with the malignant phenotype of cancer cells. Inhibition of COX-2 expression by antisense technique can reverse th

关 键 词：膀胱肿瘤 癌 核酸类 环氧合酶-2 

分 类 号：R737.14[医药卫生—肿瘤]