Fig. 6

LncRNA PTTG3P regulates the level of multiple genes involved in cell cycle, cell apoptosis and EMT through PI3K/AKT pathway in HCC cells. (a-c) The relative levels of cell cycle associated genes, including C-myc, CyclinD1, CDK6, CDK4, Rb and phosphorylated Rb (p-Rb), were detected in HepG2 cells overexpressing PTTG3P and HepG2 cells with stably suppressed PTTG3P expression by qRT-PCR (a) and western blot with quantitative analysis (b and c). β-actin was used as a housekeeping gene for qRT-PCR and an internal control for western blotting analysis. (d-e)Western blot analysis of the levels of Caspase3 and cleaved Caspase3 in HepG2 cells with silenced or enhanced PTTG3P. β-tublin was used as an internal control. (f-h) Knockdown of endogenous PTTG3P in HepG2 cells reduced the mRNA (f) and protein levels (g and h) of several EMT-marker genes including Snail and Slug but enhanced E-cadherin expression. In contrast, the level of Snail and Slug increased and the mRNA (f) and protein (g and h) level of E-cadherin decreased in HepG2 cells with elevated PTTG3P expression. β-actin was used as a housekeeping gene for qRT-PCR. β-tublin was used as an internal control for western blot analysis. (i-j) The levels of PI3K, phosphorylated PI3K (p-PI3K), AKT, phosphorylated AKT (p-AKT) were examined by western blot analysis in HepG2 cells with silenced or enhanced PTTG3P. β-actin was used as an internal control. The experiments were performed in triplicate; the data are expressed as the mean ± SD. *, P < 0.05