| miR-133b对人胃癌细胞及其靶蛋白影响的研究 |
| Alternative Title | Effect of miR-133b on human gastric cancer cell line and target protein
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| 王莉 |
| Thesis Advisor | 乔梁
; 周永宁
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| 2014-05-22
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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 | 胃癌
miR-133b
靶基因
细胞凋亡
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| Abstract | 目的：研究miR-133b在胃癌细胞系SGC-7901和BGC-823中的表达情况以及对其靶标HORMAD2、SLC6A1及HIF-1α表达的影响。
方法：利用生物信息学方法，采用microRNA靶基因预测软件预测miR-133b靶基因，且进行基因功能的初步分析；构建RNA oligo-133b转染载体，瞬时转染进SGC-7901及BGC-823胃癌细胞中，采用实时荧光定量PCR检测转然后miR-133b的表达量， MTT及流式细胞术对其功能进行验证，并采用Western Blot检测miR-133b对细胞中HORMAD2，SLC6A1及HIF-1α蛋白表达的影响。
结果：miR-133b成功转染进SGC-7901及BGC-823胃癌细胞内；qRT-PCR检测出miR-133b M转染组miR-133b表达水平较对照组升高，差异有统计学意义(P<0.05)， 转染效率很高；SGC-7901和 BGC-823 M组凋亡率升高(P<0.05)；24h和48h细胞存活率无明显差异(P>0.05)，72h和96h存活率明显下降(P<0.05)；Western Blot结果显示HORMAD2，SLC6A1在胃癌细胞中为高表达，且与HIF-1α三者均可能是miR-133b在SGC-7901及BGC-823胃癌细胞中的作用靶点，并且miR-133b M组的表达水平明显下降 (P<0.05)。
结论：1. Has-miR-133b转染胃癌细胞系SGC-7901及BGC-823后可有效表达。2. HORMAD2、SLC6A1和HIF-1α可能为miR-133b的靶基因。3. HORMAD2在人胃癌细胞系SGC-7901和BGC-823高表达，SLC6A1在BGC-823和MGC-803高表达。4. SLC6A1和HIF-1α表达下调可促进SGC-7901和BGC-823的凋亡。 |
| Other Abstract | Objective：To study the expression of Has-miR-133b in human gastric cancer cell line SGC-7901and BGC-823 and its influence on the expression of its target gene HORMAD2, SLC6A1 and HIF-1α.
Methods：Using bioinformatics methods,the target genes of miR-133b and preliminary analysis of gene function were predicted by miRNA target genes prediction software, RNA oligo-133b was structured and transfected transiently into SGC-7901 and BGC-823 gastric cancer cells, The expression of miR-133b was detected by real-time fluorescent quantitative PCR(qRT-PCR); The influence of HORMAD2, SLC6A1 and HIF-1α protein expression to miR-133b was detected by Western Blot and its function was determined by MTT and flow cytometry.
Results：miR-133b was transfected into SGC-7901 and BGC-823 successfully; the expression leve of miR-133b which was detected by qRT-PCR mimics transfection group was higher than the control group, the difference was statistically significant(P<0.05); the apoptosis rate in SGC-7901 and BGC-823 M group increased(P<0.05); cell survival rate has no obvious difference in 24 h and 48 h(P>0.05),but 72h and 96h decreased evidently(P<0.05); According to the results of Western Blot indicated HORMAD2，SLC6A1 and HIF-1α may be all of the targets of miR-133b in gastric cancer cells SGC-7901 and BGC-823, and miR-133b M group was reduced(P<0.05).
Conclusions: 1.miR-133b can effectively express in gastric cancer cell line SGC-7901 and BGC-823 after transfection. 2. HORMAD2, SLC6A1 and HIF-1α may be the target genes of miR-133b. 3. HORMAD2 is high expression in human gastric cancer SGC-7901 and BGC-823，and SLC6A1 is high expression in BGC-823 and MGC-803. 4. Cell apoptosis of SGC-7901and BGC-823 is promoted by SLC6A1 and HIF-1α lower expression. |
| 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/262010/224051
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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 |
王莉. miR-133b对人胃癌细胞及其靶蛋白影响的研究[D]. 兰州. 兰州大学,2014.
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