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目的筛选胃癌中相关miRNAs,并验证其作用靶点。方法应用基因芯片技术检测3份正常胃组织标本、24份胃癌组织标本、胃癌细胞SGC7901和正常胃黏膜细胞GES-1中328个miRNAs的表达情况。采用实时荧光定量PCR对结果进行验证,并用基因克隆和Western blot方法分析miR-9的作用靶点。结果共有26个miRNAs在胃癌标本(包括24份胃癌组织和SGC7901细胞)中异常表达。其中19个下调,7个上调。实时荧光定量PCR检测出miR-9在胃癌标本中的表达水平显著下调,该结果与基因芯片检测结果一致。miR-9与RAS癌基因家族成员RAB34的表达呈负相关。结论已初步筛选了胃癌相关miRNAs。miR-9可能是胃癌中的标记性miRNAs之一,RAB34是其作用靶点。
Objective To screen the related miRNAs in gastric cancer and validate its target. Methods The expression of 328 miRNAs in 3 normal gastric tissue specimens, 24 gastric cancer tissue specimens, gastric cancer cell line SGC7901 and normal gastric mucosal cell line GES-1 was detected by gene chip technique. The results were verified by real-time fluorescence quantitative PCR, and the target of miR-9 was analyzed by gene cloning and Western blot. Results A total of 26 miRNAs were abnormally expressed in gastric cancer specimens (including 24 gastric cancer tissues and SGC7901 cells). Among them, 19 were down and 7 were up. The expression of miR-9 in gastric cancer was significantly down-regulated by real-time fluorescence quantitative PCR, which was consistent with the results of microarray. miR-9 and RAS oncogene family member RAB34 expression was negatively correlated. Conclusion Gastric cancer related miRNAs have been initially screened. miR-9 may be one of the marker miRNAs in gastric cancer, RAB34 is its target.