2014•Chin J Hepat Surg(Electronic Edition)Requires access
Impact of multidrug resistance-related protein 3 gene silenced by siRNA on the drug resistance of liver cancer cells
Zheng Su, Bo Liu, Jianping Liu, Huayao Zhang, Zejian Lyu
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Abstract
Objective
To observe the impact of multidrug resistance-related protein (MRP) 3 gene silenced by small interfering RNA (siRNA) (MRP3-siRNA) on the drug resistance of primary liver cancer cells.
Methods
The drug resistance cell lines HepG2/Adriamycin(ADM) were developed by exposing parental cells to stepwise increasing concentrations of ADM and then MRP3-siRNAs were transfected in HepG2/ADM cells with lipofectamine 2000 liposomes. Three groups were assigned: HepG2 cell group (control group), HepG2/ADM cell group (resistance group) and MRP3-siRNA transfected HepG2/ADM cell group (interference group). The MRP3 mRNA contents of 3 groups were measured by real-time fluorescent quantitative polymerase chain reaction (PCR). The expression of MRP3 protein was detected by Western blot. The 50% inhibitory concentrations (IC50) of ADM, fluorouracil (5-FU), vincristine, oxaliplatin were determined by methyl thiazolyl tetrazolium (MTT) and the drug resistance indexes (RI) were calculated. The experimental data of 3 groups were compared by one-way analysis of variance, while pairwise comparisons were conducted using LSD-t or t test.
Results
The mean of MRP3 mRNA content in resistance group (5.16±0.31) was significantly higher than those in control group (3.08±0.27) and interference group (2.85±0.23) (LSD-t=8.765, 10.363; P<0.05). The MRP3 protein content in resistance group (21 063±274) was significantly higher than those in control group (14 476±217) and interference group (6 660±153) (LSD-t=21.836, 79.578; P<0.05). The RI of ADM, 5-FU,vincristine, oxaliplatin in resistance group were 14.40±0.31, 26.68±0.22, 28.70±0.49, 20.23±0.54, and were 3.55±0.16, 9.60±0.27, 2.11±0.17, 3.15±0.13 respectively in interference group. The RI in interference group were significantly lower than those in resistance group (t=53.873, 84.933, 88.811, 53.258; P<0.05).
Conclusions
MRP3 gene of liver cancer cells silenced by siRNA can improve the cells' sensitivity to chemotherapy drugs and reverse its chemotherapy drug resistance.
Key words:
Liver neoplasms, experimental; RNA interference; Multidrug resistance-associated proteins