| RNAi沉默galectin-1基因对肝癌细胞生物学行为的影响 |
| Alternative Title | Effects of silencing galectin-1 gene by RNAi on the behavior of hepatocyte carcinoma cells
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| 张晖汉 |
| Thesis Advisor | 张晖汉
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| 2010-05-20
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| Degree Grantor | 兰州大学
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| Place of Conferral | 兰州
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| Degree Name | 硕士
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| Keyword | 肝细胞癌
RNA干扰
细胞凋亡
肿瘤迁移
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| Abstract | 目的 建立小干扰RNA（siRNA）抑制HePG2细胞galectin-1 基因表达的实验条件，观察利用siRNA选择性抑制galectin-1基因表达对人肝癌细胞系HePG2生长、凋亡及迁移能力的影响。
方法 用RT-PCR技术检测siRNA对HePG2细胞galectin-1基因沉默的效率; 用流式细胞术检测HePG2galectin-1基因沉默前后细胞凋亡率； 以四甲基偶氮唑盐法（MTT）检测galectin-1基因沉默对HePG2细胞增殖的影响；用Millecell小室检测galectin-1基因沉默对HePG2细胞侵袭人工基底膜能力的影响。
结果 与空白对照组相比，转染48及72小时后人肝癌细胞系HePG2细胞中 galectin-1 基因表达水平明显下降（100% vs. 15.7%，12.3%，p<0.01），HePG2细胞凋亡率明显上升（0 vs. 22.5%，p<0.01），HePG2细胞增殖率显著下降（100% vs. 57.2%，p<0.01）， HePG2细胞的迁移能力明显受抑（100% vs. 53.1%，p<0.01）。
结论 基于mRAN的siRNA干扰技术可以有效抑制galectin-1 基因在人肝癌细胞系HePG2中的表达；galectin-1基因沉默对人肝癌细胞系HePG2的生物学行为具有明显的阻遏效应。由此推测，galectin-1基因表达在肝细胞癌的发生发展中起重要作用。 |
| Other Abstract | Objectives To establish the experiment condition of RNA interference to galectin-1 mRNA and investigate the roles of galectin-1 in the regulation of cell biological behaviour of hepatocyte carcinoma cells.
Methods Human hepatocyte carcinoma cells of HePG2 was cultured at 37℃ in a CO2 incubator. Special stealth RNAi was transfected to hepatocyte carcinoma cells to restrain the expression of galectin-1 mRNA. RT-PCR was used to test the silencing rate of galectin-1 gene expression.Flow cytometry was used to examine the apoptosis of HePG2 cell. The cell proliferation was examined by MTT assay and the invasion ability was tested by millicell chambers.
Results After the transfection for 48 and 72 hours, compared to the blank control, the expression of galectin-1 mRNA in HePG2 cells decreased to 15.7% and 12.3%. The cell proliferation rate was reduced to 57.2% and the number of cells undergoing apoptosis significantly increased to 22.5% after the transfection for 48 hours. Meanwhile, the galectin-1 mRNA expression restraining also resulted in an inhibition of cell migration rate to 53.1%.
Conclusion RNA interference based on siRNA can effectively inhibit galectin-1 gene expression in HePG2 cells. Silencing galectin-1 gene has an obviously deterrent effect on the cell biological behavior of HePG2 cells. The results suggested that the galectin-1 gene expression in hepatocyte carcinoma cells may play an important role in the tumor occurrence and development. |
| URL | 查看原文
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| Language | 中文
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| Document Type | 学位论文
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| Identifier | https://ir.lzu.edu.cn/handle/262010/223948
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| Collection | 学院待认领
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| Affiliation | 临床医学院
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Recommended Citation
GB/T 7714 |
张晖汉. RNAi沉默galectin-1基因对肝癌细胞生物学行为的影响[D]. 兰州. 兰州大学,2010.
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