反义整合素β6基因对结肠癌细胞作用的研究  

Effects of antisense 6 gene on colon cancer cells

作　　者：张朝阳[1] 徐克森[1] 杨广运[1] 王金申[1] 高颖 刘炎峰[1] 帅晶[1] 王家勇[1] 李少燕[3] 寿楠海[1] 牛军[1] 

机构地区：[1]山东大学齐鲁医院普外科 [2]山东省济南市医学会 [3]山东大学齐鲁医院检验科

出　　处：《中华医学杂志》2007年第37期2645-2648,共4页National Medical Journal of China

基　　金：国家自然科学基金(30570833);山东省自然科研基金(Y2005C42)

摘　　要：目的研究反义整合素β6基因对结肠癌细胞生长增殖的作用。方法先构建反义β6基因的表达载体,然后转染结肠癌细胞,使其稳定表达。应用 RT-PCR 检测细胞中β6mRNA 的表达量;应用流式细胞技术检测细胞表面β6蛋白的表达量;检测分析各组细胞与纤连蛋白结合力的改变;检测分析各组细胞增殖能力的改变;将转染细胞及正常细胞分别种植于裸鼠皮下,观察细胞的成瘤能力。将实验组与对照组的检测结果进行比较。结果与对照组相比,转染了反义β6基因的结肠癌细胞,其β6mRNA 的量及细胞表面β6蛋白的表达量(30%)均明显减少(P<0.01);其与纤连蛋白的结合力降低;其生长及增殖的能力较对照组细胞明显减弱;其在裸鼠皮下的成瘤能力明显减弱(P<0.01)。结论反义β6基因可以在转录和翻译水平明显抑制β6基因的表达,从而抑制结肠癌细胞的生长增殖,证实了整合素β6对结肠癌细胞的生长增殖具有重要的调控作用。Objective To investigate the effect of antisense integrin β6 gene on the growth of colon cancer ceils. Methods Expressing vector of antisense αvβ6 was constructed. Human colon cancer ceils of the line HT29 were cultured and divided into 3 groups ： Group A, remaining wild type ; Group B, transfected with antisense integrin β6 gene ; and Group C, transfected with blank vector. RT-PCR was used to detect the integrin β6 mRNA expression of in the HT29 ceils, The integrinβ6 protein expression on the surface of the ceils was detected by immunohistochemistry and flow cytometry, The binding between the ceils and fibronectin was examined. 3H-labeled thymidine （T） was added into the culture fluid of the ceils, and then the radiation amount was detected every 6 days so as to determine the capacity to proliferation of the ceils in vitro. Thirty female nude mice were divided into 3 groups to be injected subcutaneously with suspension of HT29 ceils of Groups A, B, and C as mentioned above, Six weeks later the size of tumors was measured and part of the tumor nodules were reseeted 5 weeks after the inoculation to undergo pathological examination. Results Compared with Groups A and C, no corresponding band at 141 bp was found in Group B by RTPCR. Flow cytometry showed that the expression level of β6 protein had was （0. 30 ± 0.051, 30% ） , significantly lower than those of Groups A and C [ （0.80 ±0.038, 80% ） and （0.85 ±0.045, 85% ）, both P 〈0.01 ]. The binding between the HT29 ceils and fibroneetin of Group B was significantly degraded after the further addition of anti-β1 and anti-αv in comparison of Groups A and C （ both P 〈 0. 01 ）, The accumulation values of 3 H-labeled T of Group B 2, 4, and 6 days after addition were all significantly lower than those of Groups A and C （all P 〈 0. 01 ）, The tumors in 9 of the 10 mice injected with the HT29 ceils of Group B disappeared and the tumor in the only one mice in Group B was only less than 1 mm^3, significantly smaller then those in Group

关 键 词：整合素β6 RNA 反义 结肠肿瘤 

分 类 号：R735.35[医药卫生—肿瘤]