Inhibitory effect of miR-34a on lungcancerstem cellsvia Notch1 signaling pathway / 中国组织工程研究

ABSTRACT

BACKGROUND:It has been proved that miR-34a plays an inhibitory role in the growth of lung cancer stem cels, but the underlying mechanism remains unclear. OBJECTIVE:To explore the inhibitory effect of miR-34a on lung cancer stem celsand the underlying mechanism. METHODS:The CD133+lung cancer stem cels were separated from lung cancer A549 cel lines using magnetic activated cel sorting method. And miR-34a-overexpressing CD133+lung cancer stem cels were established by liposome transfection technology. Besides,the targeted relationship between miR-34a and Notch1 was analyzed by the dual-luciferase reporter. Afterwards, Notch1 silencing was performed by gene knockout, and its effect on lung cancer stem cels was determined. RESULTS AND CONCLUSION:After sorted and detected by immunomagetic selection and flow cytometry assay respectively, a high rate of CD133+lung cancer stem cel was obtained. And qRT-PCR detected that the expression level of miR-34a in CD133+lung cancer stem cels was significantly lower than that in CD133-lung cancer stem cels. Moreover, miR-34a-overexpressing CD133+lung cancer stem cels were successfuly constructedandmiR-34a significantly inhibited proliferation and induced apoptosis of lung cancer stemcels. Dual-luciferase reporter assay indicated that Notch1 mRNA was a target of miR-34a. In addition, Notch1 silencing obviously inhibited proliferation and induced apoptosis of lung cancer stem cels. These findings suggest that miR-34a can inhibite lungcancer stem celsviathe Notch1 signaling pathway.