摘要
目的探讨靶向autotaxin的短发夹RNA(shRNA)对人胃癌裸鼠移植瘤的生长抑制作用。方法将已构建成功的靶向autotaxin及非特异性短发夹RNA的质粒载体pSUPER-ATX、pSUPER-mock(阴性对照)和空白质粒pSUPER-control转染胃癌AGS细胞株,并将获得的相应各组细胞与野型细胞(WT)分组种植到裸鼠皮下,以建立人胃癌裸鼠移植瘤对比模型,随后观察监测各组裸鼠的肿瘤生长情况,HE染色检测移植瘤及各器官形态学改变。第8周裸鼠处死并剥离出肿瘤组织,HE染色观察肿瘤细胞的形态,免疫组化及Western blot方法检测autotaxin蛋白及相关蛋白MMP-2、MMP-9的表达。结果各组裸鼠的内脏未受到明显的毒害和损伤。第7周pSUPER-ATX组裸鼠移植瘤的体积和质量与其余组相比均有显著性缩小(P<0.05),抑制率大约在50%左右。与野型组、阴性对照组及空白质粒组相比,pSUPER-ATX组移植瘤的autotaxin、MMP-2、MMP-9表达均有显著性下调(P<0.05)。并且autotaxin和MMP-2的表达有明显相关性(r=0.869,P<0.01)。而阴性对照组pSUPER-mock的上述指标与野型组及空白对照组相比,其差异无统计学意义。结论 所构建的靶向autotaxin的shRNA能够抑制人胃癌AGS细胞裸鼠移植瘤的生长。
Objective To investigate the effect of short hairpin RNA（shRNA） targeting autotaxin on the growth of human gastric carcinoma xenografts in nude mice. Methods Three cell clones of gastric cancer cell line AGS were stable generated, which were estab- lished successfully by transfections of the constructed plasmid vectors of autotaxin shRNA （ pSUPER - ATX） , unspecific coding shRNA （ pSUPER - mock） and the empty plasmid （ pSUPER - control）. Tumor xenograft model was generated by subcutaneous injection of AGS cells into the dorsum of nude mice. The growth of xenograft tumor was monitored and measured. Change of the tumor morphology and mice organs were observed by HE staining. The expressions of autotaxin, MMP - 2 and MMP - 9 protein in xenograft tumor tissue were detected by immunohistochemistry and western blot. Results No obvious damage of the organs was found. The pSUPER - ATX significantly sup- pressed the tumor volume and weight since the 7th week after cell transplantation, when compared to the pSUPER - mock, pSUPER - con- trol and WT groups, and the inhibition rates were about 50% （P 〈 0.05）. However, no significant difference in these parameters were found among the WT, pSUPER - mock and pSUPER - control groups. Furthermore, autotaxin, MMP - 2, MMP - 9 were expressed at sig- nificantly lower level in pSUPER - ATX group than in other three groups （P 〈 O. 05）, and a significant correlation between ATX and MMP -2 expression was displayed （r = 0. 869, P 〈 0.01 ）. Conclusion The shRNA targeting autotaxin inhibited the growth of human gastric cancer xenograft in nude mice.
出处
《医学研究杂志》
2013年第3期41-45,共5页
Journal of Medical Research
基金
浙江省自然科学基金资助项目(Y207596)
