Figure 8
Overexpression of miR-2909 in HeLa cells confirms its oncogenic properties. (A) Endogenous levels of KLF4 protein in indicated cell lines. (B) Representative fluorescent microscopic image indicating GFP expression in HeLa cells transfected with pMIRH-2909 expression vector compared with control null vector containing scrambled sequence. (C-F) Protein expression levels of KLF4 (C), P21CIP(D), MYC (E) and CCND1 (F) in HeLa cells transfected with PMIRH-2909 compared with controls. Densitometric analysis of protein bands was performed using Scion Image Analysis Software. β-actin was used as an invariant control; Each bar represents mean ± S.D. of the experiment performed in triplicate; *P < 0.05 relative to control. (G) Schematic model to summarize the critical role of miR-2909 dependent pathway in B-ALL and T-ALL. As shown, miR-2909 is functionally active and regulates KLF4 and other genes involved in cell cycle progression and apoptosis in pediatric B-ALL while its control over the regulation of KLF4 and other genes is lost because of mutations in the KLF4 3′UTR, which includes the miR-2909 binding site and the altered third zinc finger motif sequence of KLF4 in pediatric T-ALL.