MicroRNA-29a regulation SH-SY5Y cells proliferation and neurite growth through interaction with phosphatase and tensin homologue deleted on chromosome ten-phosphatidylinositol 3 kinase/protein kinase B/mammalian target of rapamycin signaling pathway
Objective To observe the effect of microRNA(miRNA)-29a and phosphatase and tensin homologue deleted on chromosome ten(PTEN)regulation on the proliferation and axon growth of nerve cells through cultured nerve cells(SH-SY5Y)in vitro.Methods Different expression levels of miRNA-29a in the cells were constructed by transfected miRNA-29a mimic and inhibitor(40 nmol/L)in vitro cultured SH-SY5Y cells(4 × 104/ml).The effects of miRNA-29a transfected(24 h after transfection)and the mRNA expression levels of PTEN,protein kinase B(Akt),and mammalian target of rapamycin(mTOR)(48 h after transfection)were detected by qPCR.The expression levels of PTEN,Akt,and mTOR protein and the phosphorylation levels of Akt and mTOR in different cells were detected by Western blotting 72 h after transfection.The proliferation activity and axonal length(μm)of different cells were measured by methyl thiazolyl tetrazolium(MTT)and neuromorphology 72 h after transfection.The data of each group were tested and compared by one-way ANOVA and LSD-t method.Results The expression level of miRNA-29a was significantly increased(9.03±0.12)in the mimic miRNA-29a group,while the expression level of PTEN gene and protein were significantly decreased(0.31±0.02,0.30±0.06),and the phosphorylation level of Akt and mTOR(3.26±0.13,2.57±0.13)in the cells with miRNA-29a high expression were also significantly increased(t=241.70,45.09,58.79,P＜0.05).The expression level of miRNA-29a was significantly decreased(0.31±0.03)in the cells transfected with inhibitor miRNA-29a,while the expression levels of PTEN gene and protein were significantly increased(3.26±0.23,3.36±0.15),and the phosphorylation levels of Akt and mTOR were significantly decreased(0.42± 0.03,0.31±0.04)in the cells with miRNA-29a depression expression(t=71.45,51.55,P＜0.05).The proliferation activity(0.86±0.07)and axonal length[(157.56±11.13)µm]were significantly in-creased in the cells with up-regulated miRNA-29a expression,which were significantly higher than those in the cells with miRNA-29a depression expression[0.44±0.03,(43.48±4.92)μm,t=13.89,31.11,P＜0.05).Conclusion MiRNA-29a can promote proliferation and axonal growth of SH-SY5Y cells by in-terfering expression of PTEN and regulating the phosphorylation levels of Akt and mTOR in the PI3K/Akt/mTOR signaling pathway.
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