miR-144在胃癌细胞系AGS中的表达及与KLF12表达相关性

The expression and correlation of miR-144 and KLF12 AGS cells

目的研究miR-144在胃癌细胞系AGS中的表达及与KLF12表达的相关性。方法应用RPMI1640培养液培养人胃癌细胞系AGS和正常胃上皮细胞系GES-1,应用qRT-PCR法检测两种细胞系中miR-144的表达,qRT-PCR和Western blot法检测KLF12的表达;在AGS细胞系中分别转染miR-144模拟物和miR-144抑制物,qRT-PCR法检测miR-144的表达,qRT-PCR和Westernblot法检测KLF12的表达。通过在线预测工具(TargetscanHuman 7.2,micro RNA.org,miRDB)预测KLF12是否为miR-144的靶基因。结果miR-144在AGS细胞系表达水平低于GES-1细胞系,而KLF12在AGS细胞系表达水平高于GES-1细胞系。AGS细胞miR-144模拟物组,miR-144的表达水平明显升高,KLF12则下降;miR-144抑制剂组中,miR-144的表达水平明显降低,而KLF12则升高。在线预测网站预测结果显示,在KLF12基因的3’UTR区域,有三处miR-144的结合位点。结论miR-144在胃癌AGS细胞系的表达与KLF12的表达呈负相关,miR-144可能通过作用于KLF12的3’UTR区域影响其表达。

Objective To study the expression of microRNA-144 in gastric cancer cell line AGS and its correlation with KLF12 expression.Methods RPMI 1640 medium was used to culture human gastric cancer cell line AGS and normal gastric epithelial cell line GES-1.The expression of microRNA-144 was detected by q-PCR.The expression of KLF12 was detected by qRT-PCR and Western blot.The mimic of microRNA-144 and the inhibitor of microRNA-144 were transfected into AGS cells respectively,qRT-PCR and Western blot were used to detect the expression of KLF12,to predict whether KLF12 is the target gene of micro RNA-144 by online prediction tools(Targets can Human 7.2,microRNA.org,microDB).Results The expression level of miR-144 was decreased in AGS cells than in GES-1 cells,but increased for KLF12.The expression level of miR-144 was significantly increased,but decreased for KLF12 in mimic group,However,miR-144 expression level was significantly decreased,but increased for KLF12 expression in inhibitor group.The online prediction website predicted that there were three binding sites of microRNA-144 in the 3’UTR region of KLF12 gene.Conclusion The expression of miR-144 in AGS cells is negatively correlated with KLF12,miR-144 affects KLF12 expression via 3’UTR region of KLF12.