Silencing microRNA-16 induces drug resistance of hepatoma cell lines via nuclear factor-κB pathway

Abstract

Objective Study on the mechanism of sliencing microRNA (miRNA, miR)-16 on chemoresistance in SMMC-7721. Methods The lentiviral vectors were used to construct stable cell lines. To determine whether constructed successfully, the expression of miR-16 was detected by real-time reverse transcription-polymerase chain reaction (RT-qPCR) and inhibitor of kappaB kinase beta (IKBKB), the target gene of miR-16, was detected by Western blotting. The sensitivity and resistance mechanism of the stable cell lines to paclitaxel were detected by Western blotting and thiazole blue (MTT). Results The expression of miR-16 in 7721-miR16 group was significantly higher than SMMC-7721 and 7721-mock groups (4.63±1.32 vs. 1.00±0.30 vs. 0.93±0.37, P =0.000). The expression of IKBKB in 7721-shmiR16 was 25.73 times than the controls (P=0.000). The expression of multidrug resistance protein 1 (MDR1) in 7721-shmiR16 was increased and the half maximal inhibitory concentration (IC50) to paclitaxel was 4.903 μmol/L, 10.64 times than the control group (0.461 μmol/L) (P=0.000). The results of western showed that silencing miR-16 induced resistance mainly through activating nuclear factor-kappaB (NF-κB) pathway. Conclusion Silencing miR-16 in SMMC-7721 significantly increases IKBKB expression leadind paclitaxel resistance. Key words: Hepatoma; MicroRNA-16; Paclitaxel; Drug resistance; Inhibitor of kappaB kinase beta