Role and Mechanism of miR-1269a on Targeted Regulation of HOXD10 in Invasion of Cholangiocarcinoma Cells

Objective To explore the possible mechanism of microRNA-1269a(miR-1269a) on the targeted regulation of HOXD10 in the invasion of human cholangiocarcinoma (CCC) cells.

Methods We detected six miRNAs levels in human CCC samples and cells for screening miR-1269a as the research object.miR-1269a mimic and inhibitor were transfected into four CCC cells by Lipofectamine liposome respectively.The expressions of HOXD10 mRNA and protein were detected by real-time quantitative PCR (qPCR) and Western blot.The effect of miR-1269a on the invasion of CCC cells was observed.Double luciferase reporter assay was applied to verify the targeting relationship between miR-1269a and HOXD10.

Results miR-1269a expression was significantly upregulated in CCC tissues, compared with adjacent normal tissues (P=0.0023).The mRNA and protein levels of HOXD10 in miR-1269a mimic transfection group were lower than those in control group (Mz-CHA-1:P=0.0025;RBE:P=0.0038).miR-1269a mimic significantly elevated the invasion capacity of CCC cells (Mz-CHA-1:P=0.004;RBE:P=0.004), while miR-1269a inhibitor remarkably inhibited the invasion (QBC939:P=0.16;HCCC9810:P=0.13).Double luciferase reporter gene test showed that miR-1269a could significantly inhibit the luciferase activity of wild-type HOXD10-3'UTR, but had no effect on the luciferase activity of mutant plasmid transfected cells.

Conclusion  miR-1269a may regulate the invasion of CCC cells by targeting HOXD10, and could be used as an effective target for the molecular therapy of CCC.