Survivin反义核酸治疗人大肠癌裸鼠皮下种植瘤实验探究宫向前.docVIP

Survivin反义核酸治疗人大肠癌裸鼠皮下种植瘤实验探究宫向前 　 作者：乔广东 李玉华 邢建华 寿楠海 丁印鲁 【摘要】目的：探讨Survivin反义寡核苷酸对裸鼠人大肠癌皮下种植瘤体生长的抑制作用及其机制。方法：接种人大肠癌细胞制作裸鼠皮下大肠癌种植瘤模型，分别用Survivin反义寡核苷酸、正义核苷酸和生理盐水对皮下瘤体内直接进行注射治疗，检测肿瘤的生长状况、瘤体体积及瘤重变化，计算抑制率。病理切片免疫组化检测瘤体ECD表达。结果：经Survivin硫代反义寡核苷酸治疗组裸鼠皮下移植瘤生长受到抑制，肿瘤重量明显小于对照组，肿瘤体积明显小于对照组和正义组（P＜0.05），裸鼠皮下移植瘤ECD表达增强。结论：Survivin反义核酸可以抑制大肠癌裸鼠移植瘤的生长，Survivin反义核酸抑制大肠癌裸鼠移植瘤生长的机理可能与ECD基因表达变化有关。 【关键词】寡核苷酸类，反义#12539;基因，Survivin#12539;结直肠肿瘤#12539;小鼠，裸 【ABSTRACT】Objective:To study the inhibitory effect of survivin antisense oligodeoxynucleotides on growth of human colorectal carcinoma in nude mice.Methods:Human colorectal carcinoma transplanted subcutaneously in nude mice model was estabished.A total of 24 mice were divided into three groups,namely control group,survivinSODN group and survivinASODN group.Different treatments were given respectively,continuous infusions over 16 days.The weight and volume of the subcutaneous tumors was measured,and the morphology of tumor was observed.The expression of Ecadherin was examined immunohistochemically in the human colorectal cancer transplanted subcutaneously in nude mice.Results:The growth inhibitory rate was 57%in ASODN group,while only 6.67% was in SODN group.The weight and volume of the subcutaneous tumors in survivinASODN group is smaller than in control group.And the expression of Ecadherin is stonger in the ASDON group than in control group.Conclusion:Survivin antisense oligodeoxynucleotides are powerful of inhibiting the growth of human colorectal carcinoma in nude mice.The change of Ecadherin,another tumorrelated gene,may play an important role in this progress. 【KEY WORDS】Oligodeoxynucleotides antisense#12539;Genes,survivin#12539;Colorectal neoplasms#12539;Mice,node 研究表明，肿瘤的发生发展与基因缺失、突变或异常表达有关，因此基因疗法为肿瘤治疗提供了新的途径［1］。我们以Survivin基因为靶基因，探讨了其反义寡核苷酸对裸鼠人大肠癌生长的抑制作用及机制。 1 材料与方法 1.1 细胞培养 HT29结肠腺癌细胞株为中分化Ⅱ级上皮样细胞。于含10%小牛血清的RPMI1640培养液（PH7.27.4,Gibico,U.S.A)培养瓶中培养，内含青霉素和链霉素各100U/ml，单层培养于37℃，5%CO2饱和湿度培养箱中。0.25%胰蛋白酶消化，传代后处于指数生长期备用。实验用细胞经0.4%台盼