Wang Zheng,Zheng Lu,Huang Xiaobing,Yang Tonghan,Li Jing,Liang Ping.Mechanism underlying drug resistance of HepG2/ADM cells silently reversed by lentivirus-mediated BC047440 gene during chemotherapy[J].J Amry Med Univ (J Third Mil Med Univ),2012,34(08):705-709.
Wang Zheng,Zheng Lu,Huang Xiaobing,Yang Tonghan,Li Jing,Liang Ping.Mechanism underlying drug resistance of HepG2/ADM cells silently reversed by lentivirus-mediated BC047440 gene during chemotherapy[J].J Amry Med Univ (J Third Mil Med Univ),2012,34(08):705-709.DOI:
Mechanism underlying drug resistance of HepG2/ADM cells silently reversed by lentivirus-mediated BC047440 gene during chemotherapy
Objective To observe the effect of RNA interference (RNAi)-silenced BC047440 gene on multidrug resistance of HepG2/ADM cells and its molecular mechanism. Methods BC047440 protein siRNA interference model was established in Hep
G2/ADM cells. Expression of BC047440 protein was detected by Western blotting. Experiment was conducted in BC047440-shRNA group
control-shRNA group and HepG2/ADM group. Effect of adriamycin on cell growth was analyzed with CCK-8 method. Cell cycle and apoptosis were assayed by flow cytometry. Expression of NF-κB protein
Survivin mRNA and CCNL1 mRNA was detected by Western blotting and RT-PCR
respectively
after BC047440 was silenced. Results The siRNA interference model was successfully established. The NF-κB protein was expressed about 71.5% in BC047440-shRNA group and HepG2/ADM group
and about 70.0% in control-shRNA group. The cytotoxicity of adriamycin at 24 and 48 h was significantly higher in BC047440-shRNA group. Flow cytometry showed that the number of apoptotic cells was significantly greater in BC047440-shRNA group
the cell cycle was maintained at the G
14
/G
0
phase
and the number of HepG2/ADM cells was reduced at the S phase. RT-PCR showed that the Survivin mRNA was expressed about 37% in control-shRNA group and about 42% in HepG2/ADM group
while the expression of CCNL1 mRNA was about 3.5-fold higher in BC047440-shRNA group than in control-shRNA group and about 2.4-fold higher than in HepG2/ADM group. Conclusion Silencing BC047440 gene can reverse the multidrug resistance of HepG2/ADM cells though the NF-κB
and downstream survivin and CCNL1 signaling pathways
which indicates that BC047440 gene may be one of the important molecules for the multidrug resistance of liver cancer.