论文标题:反义寡核苷酸抑制β-catenin 及 C-myc基因对肝癌细胞VEGF表达影响的研究
Study of Inhibitory Effects of Antisense Oligonucleotides on b-Catenin c-myc VEGF Gene Expression in Human Hepatocellular Carcinoma Cell
论文作者 周颖
论文导师 杜芳腾,论文学位 硕士,论文专业 消化内科
论文单位 江西医学院,点击次数 107,论文页数 37页File Size472k
2005-05-01论文网 http://www.lw23.com/lunwen_726692772/ 反义寡核苷酸;肝癌;β-catenin;C-myc;VEGF
Antisense oligodeoxynucleotide; Hepatocellular carcinoma; β-catenin; C-myc; VEGF
目的: 研究针对β-catenin、C-myc 基因的反义寡核苷酸对VEGF 表达水平的影响,探讨SMMC7721 肝癌细胞株中VEGF 的表达是否受到C-myc 基因的调控以及针对β-catenin、C-myc 的反义基因治疗在肝癌基因治疗中的应用前景。方法:(1)常规培养肝癌SMMC7721 细胞株并接种于6 孔板。(2)根据Genebank 收录的β-catenin、C-myc mRNA 序列设计针对翻译起始部位的反义寡核苷酸。(3)分别用针对β-catenin 翻译起始部位的反义、错配寡核苷酸以及针对C-myc 翻译起始部位的反义、错配寡核苷酸处理细胞,同时设空白对照组,缺氧条件下培养24 小时后收集细胞提取总RNA。(4)用RT-PCR 方法及FR200 图像分析系统半定量分析β-catenin、C-myc 及VEGF 的相对表达量,以β-actin表达为各基因表达的内对照。(5)应用X2检验进行数据统计学分析。结果: (1)针对β-catenin 翻译起始部位的反义寡核苷酸明显抑制β-catenin mRNA的表达,反义组和错义组抑制率分别为0.73±0.06 和0.10±0.07。同时VEGF 的表达也受到了抑制,VEGF 的抑制率为0.80±0.05 和0.09±0.06。(2)针对C-myc 翻译起始部位的反义寡核苷酸抑制C-myc mRNA 的表达,反义组和错义组抑制率分别为0.75±0.04和0.11±0.05,同时VEGF的表达也受到了抑制,VEGF 的抑制率分别为: 0.60±0.07 和0.08±0.03。结论:(1)我们根据Genebank 收录的β-catenin、C-myc mRNA
Objective: Studying the inhibitory effects of antisenseoligonucleotides on β-catenin 、C-myc and VEGF gene expression inhuman hepatocellular carcinoma cell, To investigate whether VEGF isregulated by C-myc .METHODS: SMMC7721 cells were cultured under hypoxicconditions for 24h, followed by being transfected with antisenseoligonucleotides 、scramble sequence on β-catenin 、c-myc gene. Thetotal RNAs were extracted from the cells, and the expression ofβ-catenin 、C-myc and VEGF were examined by reversetranscription-polymerase chain reaction(RT-PCR)and FR200 analysissystem , using X2 text as the statistics analysis method.RESULTS : The ASODN had obviously sequence-specificinhibitory effect on β-catenin gene expression, with the ratio on 0.73±0.06 and 0.10±0.07 on mRNA level, The expressions of VEGF geneshowed to be inhibited, with the ratio 0.80±0.05 and 0.09±0.06 . TheASODN had inhibitory effect on C-myc gene expression,with the ratioon 0.75±0.04 and 0.11±0.05. The expressions of VEGF gene showed tobe inhibited too, with the ratio on 0.60±0.07 and 0.08±0.03.CONCLUSION: (1)Our ASODNs designed for β-catenin gene and C-myc gene can inhibit effectively the mRNA expression of β-catenin gene 、C-myc gene and VEGF gene. (2)Comparing withthe ASODN designed for C-myc gene, the ASODN designed forβ-catenin gene can reduce the expression of VEGF more strongly .
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