Objective: This study explored the ability of microRNA-135a (miR-135a) to influence cell proliferation, migration, invasion, apoptosis and tumor angiogenesis through the IGF-1/PI3K/Akt signaling pathway in non-small cell lung cancer (NSCLC). Methods: NSCLC tissues and adjacent normal tissues were collected from 138 NSCLC patients. Quantitative real-time polymerase chain reaction (qRT-PCR) was used to detect the expression levels of miR-135a and IGF-1, PI3K, Akt, VEGF, bFGF and IL-8 mRNA; western blotting was used to determine the expression levels of IGF-1, PI3K and Akt protein; and enzyme-linked immunosorbent assay (ELISA) was used to analyze the expression levels of VEGF, bFGF and IL-8 protein. Human NSCLC cell lines (A549, H460, and H1299) and the human bronchial epithelial cell line (HBE) were selected. A549 cells were assigned to blank, negative control (NC), miR-135a mimics, miR-135a inhibitors, IGF-1 siRNA and miR-135a inhibitors + IGF-1 siRNA groups. The following were performed: an MTT assay to assess cell proliferation, a scratch test to detect cell migration, a Transwell assay to measure cell invasion, and a flow cytometry to analyze cell apoptosis. Results: The expression level of miR-135a was lower while those of IGF-1, PI3K and Akt mRNA were higher in NSCLC tissues than in the adjacent normal tissues. Dual-luciferase reporter assay indicated IGF-1 as a target of miR-135a. The in vitro results showed that compared with the blank group, cell proliferation, migration and invasion were suppressed, mRNA and protein levels of IGF-1, PI3K, Akt, VEGF, bFGF and IL-8 were reduced, and cell apoptosis was enhanced in the miR-135a mimics and IGF-1 siRNA groups. Compared with the IGF-1 siRNA group, cells in the miR-135a inhibitors + IGF-1 siRNA group demonstrated increased cell proliferation, migration and invasion, elevated mRNA and protein levels of IGF-1, PI3K, Akt, VEGF, bFGF and IL-8 and reduced cell apoptosis. Conclusion: These findings indicated that miR-135a promotes cell apoptosis and inhibits cell proliferation, migration, invasion and tumor angiogenesis by targeting IGF-1 gene through the IGF-1/PI3K/Akt signaling pathway in NSCLC.

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a b s t r a c tDuring direct laser deposition process, rapid melting-solidification and addition of multicomponent powder lead to complex transport phenomena in the melt pool. The thermal behavior and mass transport significantly affect the solidified microstructure and properties of fabricated layer. In this paper, an improved 3D numerical model is proposed to simulate the heat transfer, fluid flow, solidification and multicomponent mass transport in direct laser deposition of Co-base alloy on steel. The solidification characteristics, including temperature gradient (G), solidification growth rate (R) and cooing rate (G Â R), can be obtained by transient thermal distribution to predict the morphology and scale of the solidification microstructure. Multicomponent transport equation based on a mixture-averaged approach is combined with other conservation equations. The calculated melt pool geometry and the composition profiles of iron (Fe), carbon (C), cobalt (Co) and chromium (Cr) are compared with the experimental results. The results show that in the initial stage of direct laser deposition, the rapidly mixture of substrate material and added material occur in the melt pool and conduct plays an important role in heat transfer due to the low Peclet number. As the melt pool is developed, the heat and mass transfer in the melt pool are dominated by strong Marangoni convection. An unmixed zone is observed near the bottom of melt pool where the convection is frictionally dissipated due to the presence of solidified dendrites. Since the G/R decreases and G Â R increases from the bottom to the top of the solidified track, the morphology of the microstructure changes from planar front to columnar dendrites to equiaxed dendrites and the grain size decreases.

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