胃癌相关miRNAs的筛选及其作用靶点的验证

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。

Screening of Gastric Cancer-associated miRNAs and Verification of Its Target

Objective To screen gastric cancer-associated miRNAs and verify its target. Methods Three normal gastric tis-sue and 24 gastric cancer tissue specimens as well as gastric cancer SGC7901 cells and normal gastric mucosa GES-1 cells were test-ed for expression of 328 miRNAs by microarray technique, and the results were verified by real-time fluorescent quantitative PCR. The target of miR-9 was analyzed by gene cloning and Western blot. Results The expressions of 26 miRNAs were abnormal in gas-tric tissue specimens and SGC7901 cells, of which 19 were down-regulated and 7 were up-regulated. Real-time fluorescent quantita-tive PCR proved that the expression of miR-9 in gastric cancer tissue specimens was down-regulated significantly, which was consistent with the test result by microarray technique. miR-9 was negatively related to the expression of RAB34, a member of RAS gene family. Conclusion Gastric cancer-associated miRNAs were screened primarily, of which miR-9 targeting RAB34 might be one of the marker miRNAs in gastric cancer.