Effect of microRNA-135a on Cell Proliferation, Migration, Invasion, Apoptosis and Tumor Angiogenesis Through the IGF-1/PI3K/Akt Signaling Pathway in Non-Small Cell Lung Cancer.
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).
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.
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.
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.
Zhou Y
，Li S
，Li J
，Wang D
，Li Q
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Upregulation of lncRNA FER1L4 suppresses the proliferation and migration of the hepatocellular carcinoma via regulating PI3K/AKT signal pathway.
This study aimed to investigate the potential function of FER1L4 in the progression of hepatocellular carcinoma and uncover its underlying molecular mechanism.
In the current study, quantitative real-time polymerase chain reaction (qRT-PCR) was conducted to determine the expression profile of FER1L4 in normal liver tissues and hepatocellular carcinoma tissues of human, as well as hepatocellular carcinoma (HCC) cell lines including HL-7702[L-02], HepG-2, Hep3b, and SMMC-7721. Then, HepG-2 cells were transfected with pcDNA3.1-FER1L4 (pcDNA3.1-empty as negative control) for gain-of-function analysis, followed with cell functional abnormality tests. Specifically, colony formation analysis and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide experiment were taken advantage to measure the cell proliferation, while cell migration and invasion were evaluated by wound healing assay and transwell experiment respectively. Additionally, cell apoptosis was detected by flow cytometry. Moreover, the effect of FER1L4 on PI3K/AKT signal pathway activation was investigated through analyzing phosphorylation of related proteins, p-AKT/AKT and p-PI3K/PI3K, via Western blot assay.
Downregulation of FER1L4 in hepatocellular carcinoma tissues and cells was demonstrated by qRT-PCR analysis. Besides, FER1L4 overexpression evidently attenuated the cell proliferation, migration and invasion, but prompted cell apoptosis. Importantly, Western blot assays revealed that PII3K/AKT signal pathway were involved in mediating the progression regulation role of FER1L4 in HCC cells.
Our study suggested that FER1L4 might alleviate progression of hepatocellular carcinoma via blocking PI3K/AKT pathway, which encourages a better understanding of the pathogenesis of HCC and may provide a novel potential therapeutic target for clinical treatment.
Wang X
，Dong K
，Jin Q
，Ma Y
，Yin S
，Wang S
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ResearchGate
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