环氧合酶-2反义寡核苷酸对Colo-16细胞增殖及环氧合酶-2表达的影响  

In vitro effect of cyclooxygenase （COX）-2 antisense oligonucleotide on the proliferation and expression of COX-2 in human skin squamous cell carcinoma cell line Colo-16

作　　者：马杰[1] 魏志平[1] 刘彦群[1] 

机构地区：[1]江苏徐州医学院附属医院,221002

出　　处：《中华皮肤科杂志》2009年第12期835-838,共4页Chinese Journal of Dermatology

摘　　要：目的研究环氧合酶（COX）-2反义寡核苷酸（AsODN）对人皮肤鳞状细胞癌细胞株Colo-16细胞体外增殖的抑制作用及COX一2表达的影响。方法人工合成COX-2AsODN，利用脂质体将不同浓度（50，100，200，400nmol／L）反义寡核苷酸转染入Colo-16细胞，免疫荧光显微镜观察转染情况；采用噻唑蓝（MTT）法检On,4Colo-16细胞的增殖情况；Western印迹及半定量逆转录PCR检测Colo-16细胞COX-2蛋白和mRNA表达水平。结果不同浓度的COX-2AsODN对Colo-16细胞体外增殖均具有抑制作用（P〈0．05），400nmol／L反义组在48h抑制率最高，达60．3％；COX-2AsODN转染Colo-16细胞后，COX-2蛋白和mRNA表达明显下调，50、100、200、400nmol／L组COX-2蛋白灰度值分别为0．763±0．070、0．600±0．065、0．430±O．074、0．251±0．045，与阴性对照组相比差异均有统计学意义（P〈0．05）；COX-2mRNA表达量随COX-2AsODN浓度的增加而减少，50、100、200、400nmol／L组COX-2mRNA灰度值分别是0．778±0．025、0．602±0．041、0．417±0．031、0．297±0．051，各组间COX一2mRNA表达差异有统计学意义（F：132．48，P〈0．01）。结论COX-2AsODN对Colo-16细胞的体外增殖有抑制作用，并能下调COX-2蛋白及其mRNA在Colo-16细胞中的表达，提示COX-2AsODN有潜在治疗皮肤鳞状细胞癌的作用。Objective To investigate the effects of COX-2 antisense oligonucleotide （AsODN） on the proliferation and expression of COX-2 in human skin squamous cell carcinoma cell line Colo-16. Methods The COX-2 AsODN was synthesized artificially, and various concentrations （50, 100, 200, 400 nmol/L） of the AsODN were transfected into Colo-16 cells with lipofectin followed by additional culture for different durations. The transfection results were observed with fluorescence microscopy. Subsequently, MTT assay, Western blotting and reverse transcription PCR were used to detect the cell proliferation, protein and mRNA expression of COX-2 in Colo-16 cells, respectively. Results Compared with untreated cells, the proliferation of Colo-16 cells was inhibited significantly at 24, 48, 72 and 96 hours after transfection with different concentrations of COX-2 AsODN （all P 〈 0.05 ）, and the COX-2 AsODN of 400 nmol/L exerted the highest inhibition rate of 60.3% at 48 hour. The average gray scale was 0.763± 0.070, 0.600 ± 0.065, 0.430± 0.074 and 0.251 ±0.045 for COX-2 protein, 0.778 ±0.025, 0.602 ± 0.041, 0.417± 0.031 and 0.297 ± 0.051 for COX-2 mRNA in Colo-16 cells transfected with COX-2 AsODN of 50, 100, 200, and 400 nmol/L respectively, significantly lower than that in untreated Colo-16 cells （all P 〈 0.05 ）; there was a significant difference in the expression of COX-2 protein and mRNA among the cells transfected with the four concentrations of COX-2 AsODN and untreated cells （F = 83.54, 132.48, respectively, both P 〈 0.05）. Conclusions COX-2 AsODN can inhibit the proliferation, as well as the expression of COX-2 protein and mRNA in Colo-16 cells, which suggests that COX-2 AsODN has a potential therapeutic effect on skin squamous cell carcinoma.

关 键 词：癌 鳞状细胞 环氧合酶2 寡核苷酸类 反义 细胞系 肿瘤 Colo-16细胞 

分 类 号：R739.5[医药卫生—肿瘤]