胃癌与癌旁组织miRNA差异表达谱的研究  

Differential Expression of miRNAs in Human Gastric Cancer Versus Non-Tumor Adjacent Tissues

作　　者：张岱[1] 王振宁[2] 罗阳[3] 徐岩[2] 刘彦山[1] 杨威[1] 张学[1,3] 

机构地区：[1]中国医学科学院基础医学研究所医学遗传学系医学分子生物学国家重点实验室,北京市100005 [2]中国医科大学附属第一医院肿瘤外科,沈阳市110001 [3]中国医科大学基础医学院医学基因组学教研室卫生部细胞生物学重点实验室,沈阳市110001

出　　处：《医学分子生物学杂志》2010年第4期283-290,共8页Journal of Medical Molecular Biology

基　　金：资助项目：国家杰出青年科学基金（No.30125017）,国家自然科学基金（No.30370640）,国家重点基础研究发展规划项目（973计划）（No.G1998051203）

摘　　要：目的研究胃癌中表达失调的miRNA及其生物学功能，从而进一步阐明miRNA在胃癌发生中的分子机制。方法将总RNA加ploy（A）尾后进行反转录PCR扩增特异miRNA，使用QuantityOne软件进行定量分析，计算每对样本胃癌与癌旁组织比值（T／NRatio），使用SAM软件进行统计分析。MTT法检测miR-21和miR-17-5p对胃癌细胞系生长的影响。结果在8对胃癌及癌旁组织样本中对237个miRNA进行了表达谱分析。对于检测到表达的161个miRNA，使用SAM软件进行统计分析，确认22个在胃癌中表达上调，2个在胃癌中表达下调（FDR=0．0963）。进一步通过生长抑制试验证实在胃癌组织中表达异常增高的miR-21和miR-17-5p对胃癌细胞的生长有明显的促进作用。结论这些在胃癌中异常表达的miRNAs具有成为新一代胃癌标记物的潜力，能够为胃癌的精确诊断分型提供依据，同时针对这些靶点可以开发新的核酸治疗技术，通过抑制或增强其功能来达到治疗胃癌的目的。Objective To investigate the miRNA expression profiles of gastric cancer, and try to unravel the biological mechanism of certain deregulated miRNAs on gastric tumorigenesis. Meth- ods Total RNA was polyadenylated by poly （A） polymerase, and then eDNA was synthesized by a specific reverse transcription （RT） primer and reverse transeriptase using the poly （A） -tailed total RNA as templates. Then, we used QuantityOne software to calculate the ratio between each pair of gastric tumors vs. non-tumor adjacent tissues （ T/N Ratio） to evaluate miRNA expression profiles, and analyzed the data by SAM. In addition, the knockdowns of the miR-21 and miR-17- 5p on gastric cancer cell lines were detected by MTT assay. Results We identified 22 overexpressed miRNAs and 2 decreased miRNAs in our 8 paired match gastric cancer specimens by SAM （FDR = 0. 0963）, from total 237 miRNAs panel with 161 miRNAs detected whereas other 76 miRNAs undetected. Knockdown of miR-21 and miR-17-5p in gastric cancer cell lines resulted in growth inhibition, which suggested that aberrantly expressed miR-21 and miR-17-5p may contribute to gastric tumorigenesis. Conclusion Our miRNA expression results reported here increase our understanding of the molecular basis of human gastric cancer and suggest that aberrant expression of miRNA genes may be important to the pathogenesis of this human neoplasm. New therapeutics will be developed to improve clinical outcomes of gastric cancer patients based on these targets.

关 键 词：胃癌 MIRNA 表达谱 miR-17-5p MIR-21 

分 类 号：R394.3[医药卫生—医学遗传学]