Objective      To study the mechanism of miR-4686 regulating the proliferation of A549 spheres.       Methods      CD133

+

CD44

+

A549 cells were sorted by FACS

and then cultured in serum-free medium. Positive rate of CD133

+

CD44

+

A549 spheres were detected by FACS. Wnt5a and Wnt2 protein expression of A549 cells and A549 spheres were assayed by Western blotting. MiR-4686 inhibitors and negative co

ntrol were synthesized. A549 spheres were treated with miR-4686 inhibitors and negative control separately for 48 h

and then the expression of miR-4686 were detected by real-time PCR. Wnt5a

Wnt2 and β-catenin mRNA and protein expression was assayed by RT-PCR and Western blotting

respectively. The proliferation of A549 spheres was detected by MTT assay at 12

24 and 48 h.       Results      CD133

+

CD44

+

A549 cells were sorted. The positive rate of CD133

+

CD44

+

was 69.800% in A549 spheres. Wnt5a and Wnt2 protein expression was significant decreased in A549 cells

as compared to A549 spheres (

P

0.05). After treatment with miR-4686 inhibitors

the expression of miR-4686 as well as the mRNA and protein expression of Wnt5a

Wnt2 and β-catenin were significant down-regulated

compared to the control group and miR-4686 negative control group (P0.05). What’s more

OD values at 12

24 and 48 h were decreased after miR-4686 inhibitor treatment (

P

0.05).       Conclusion      MiR-4686 plays an important role in regulation of A549 sphere proliferation through Wnt signal pathway

which may become a novel target for treatment of non-small-cell lung cancer.