GAS5 Regulates RECK Expression and Inhibits Invasion Potential of HCC Cells by Sponging miR-135b

Figure 3

GAS5 regulates miR-135b expression and is a direct target of miR-135b. (a) Huh-7 and HepG2 cells were infected by lentivirus (LV) expressing GAS5. Huh-7 and HepG2 cells were transfected with GAS5 siRNAs. 72h later, the expression of miR-135b was detected by qRT-PCR. (b) Putative miR-135b binding sites in GAS5 lncRNA sequence by starBase v2.0 online software. Wild-type (WT) binding sequences were highlighted in green, while mutant (MT) binding sequences were highlighted in red. (c) Huh-7 and HepG2 cells were cotransfected with wild-type or mutant GAS5 reporters and miR-135b overexpressing or control lentivirus. 72h later, cells were harvested and firefly luciferase activity was normalized to Renilla luciferase activity. (d) Huh-7 and HepG2 cells were infected by lentivirus (LV) expressing miR-135b. 72h later, the expression of GAS5 was detected by qRT-PCR. (e) Huh-7 and HepG2 cells were infected by lentivirus (LV) expressing GAS5, without or with miR-135b-LV. 72h later, the expression of miR-135b was detected by qRT-PCR. (f) Cell invasion assay was performed to investigate the invasion ability of miR-135b in GAS5-overexpressing cells as described in (e). Data represent the mean±SD from three independent experiments. : P<0.05, : P<0.01, and NS: no significance. Scale bar: 50μm.

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