Yanfei Cao, Rui Ren, Ye Yang, et al. MicroRNA-185 inhibits proliferation and invasion of lung squamous cells via regulating E2F6. [J]. J Third Mil Med Univ 39(2):123-129(2017)
DOI:
Yanfei Cao, Rui Ren, Ye Yang, et al. MicroRNA-185 inhibits proliferation and invasion of lung squamous cells via regulating E2F6. [J]. J Third Mil Med Univ 39(2):123-129(2017) DOI: 10.16016/j.1000-5404.201607118.
MicroRNA-185 inhibits proliferation and invasion of lung squamous cells via regulating E2F6
To determine the effect of microRNA-185 (miR-185) on the proliferation and invasion of lung squamous cells and investigate the possible underlying mechanism.
Methods
2
Human lung squamous cancer cell lines (NCI-H520 and NCI-H226) and normal lung cell line (BEAS-2B) were used in the research. The miR-185 expression was detected by quantitative real-time PCR (qRT-PCR) in the above cells. The targeted relationship between miR-185 and E2F6 was detected by dual luciferase reporter assay. After MiRNA-NC and miR-185 mimics were transfected into the cells by using Turbofect
the protein expression of E2F6 was measured by Western blot assay
cell proliferation was detected by MTT assay
and cells invasion was measured by cell invasion kit. pcDNA.3.1-E2F6 recombinant plasmid was constructed
and transfected into lung squamous cells to over-express E2F6.
Results
2
miR-185 expression was significantly decreased in the lung squamous cells (
P
<0.01). miR-185 directly targeted E2F6 gene (
P
<0.05). miR-185 mimics transfection markedly up-regulated the content of miR-185 (
P
<0.01) and inhibited the protein expression of E2F6 (
P
<0.01)
and obviously suppressed the cell proliferation (
P
<0.05 or
P
<0.01) and invasion (
P
<0.05) of lung squamous cells. Transfection of pcDNA.3.1-E2F6 resulted in the over-expression of miR-185
the up-regulation of E2F6 protein (
P
<0.01)
and thus significantly promoted the cell proliferation (
P
<0.01) and invasion (
P
<0.05).
Conclusion
2
miR-185 inhibits the proliferation and invasion of the lung squamous cells
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