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不同转移潜能人大细胞肺癌细胞株转移相关microRNAs的筛选研究

ISSN号：1009-3419
期刊名称：《中国肺癌杂志》
时间：0
分类：R734.2[医药卫生—肿瘤;医药卫生—临床医学]
作者机构：[1]四川大学华西医院,成都610041, [2]天津医科大学总医院天津市肺癌研究所天津市肺癌转移与肿瘤微环境重点实验室,天津300052
相关基金：国家自然科学基金重点项目（No.30430300）; 国家“863”重大项目（No.2006AAOZA401）; 国家“973”重大项目（No.2010CB529405）; 天津市科技支撑计划重点项目（No.06YFSZSF05300）; 天津市创新体系建设项目（No.07SYSYSF05000）资助

作者：鲁为山[1], 李书军[2], 刘彬[2], 李洋[2], 罗猛[1], 孙丽亚[2], 尤嘉琮[2], 周清华[1,2]

关键词：肺肿瘤, 细胞系, 转移, 微小RNA, Lung neoplasms, Cell line, Metastasis, MicroRNAs

中文摘要：

背景与目的微小RNA（microRNAs,miRNAs）参与调节肿瘤发生发展的多个过程,包括细胞的分裂增殖、细胞周期、凋亡、血管形成、侵袭和转移等。本研究应用miRNA芯片检测具有高低不同转移潜能人大细胞肺癌细胞株L9981和NL9980的miRNA表达谱,从中筛选出与大细胞肺癌转移相关的miRNAs。方法收集L9981和NL9980细胞,抽提总RNA进行CY3标记,将标记RNA在miRNA芯片上进行杂交反应。通过数据统计分析,筛选出表达明显差异的miRNAs。应用Real-timePCR验证芯片结果,并应用生物信息学方法预测靶基因。结果在不同转移潜能人大细胞肺癌L9981和NL9980细胞株中共筛选到22个表达明显差异的miRNAs。与NL9980相比,在L9981中有13个miRNAs表达上调,9个表达下调。Real-timePCR验证miR-125a-3p在细胞中的表达水平与芯片结果趋势一致,预测其靶基因可能为胰岛素样生长因子2。结论筛选得到与大细胞肺癌转移相关的miRNA表达谱。

英文摘要：

Background and objective MicroRNAs （miRNAs） regulate a wide range of cancer-associated processes, including cell division, proliferation, cell cycle, apoptosis, angiogenesis, invasion, and metastasis. A microarray was performed to analyze metastasis-related miRNAs with di erent metastastic potentials and to further elucidate their mechanism in the large-cell lung cancer cell lines. Methods L9981 and NL9980 cells were harvested, and total RNA was extracted for CY3. RNA hybridization was then performed on the chip with marked miRNAs. MiRNAs with signi cantly di erent expression were selected through statistical analysis. A real-time polymerase chain reaction （PCR） was employed to validate the results of the microarray, and target genes were predicted using bioinformatics. Results Expressions of 22 miRNAs were signi cantly di erent in the L9981/NL9980 cell lines. Compared with the NL9980, 13 miRNAs were upregulated in the L9981 cell lines, whereas 9 miRNAs were downregulated. e result of miR-125a-3p expression based on real-time PCR was consistent with the microarray. Insulin-like growth factor 2 might be a target gene of miR-125a-3p. Conclusion e metastatic miRNA pro le in large-cell lung cancer was successfully screened out.

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