Cisplatin induces hepatitis B virus reactivation via CREB1： An experimental study

Abstract:

Objective：To investigate the mechanism of cisplatin-induced hepatitis B virus（HBV） reactivation. Methods：HepAD38 hepatoma cells with stable expression of HBV were divided into phosphate buffered saline（PBS） control group and cisplatin treatment group. The expression of cAMP responsive element binding protein 1（CREB1） and HBV proteins（HBsAg and HBcAg） and the level of HBV replication（HBV DNA and HBV mRNA） were mea－sured. HepAD38 cells with stable knock-down of CREB1（shCREB1） were compared with those with stable expression of HBV，and the level of HBV replication（HBV DNA and HBV mRNA） and protein expression level after cisplatin treatment were measured. Results：As for HepAD38 hepatoma cells with stable expression of HBV，Western blot showed that compared with the PBS control group，the cisplatin treatment group had a relative expression level of CREB1 of 1.355±0.049（P=0.010），a relative expression level of HBsAg protein of 1.679±0.060（P=0.003），and a relative expression level of HBcAg protein of 1.488±0.047（P=0.005）. The results of qRT-PCR showed that com－pared with the PBS control group，the cisplatin treatment group had an absolute expression level of HBV DNA of 249.600±54.400（P=0.006） and a relative expression level of HBV mRNA of 3.084±0.256（P=0.000），suggesting that compared with the PBS control group，the cisplatin treatment group had significantly higher expression levels of HBV proteins and level of HBV replication. In addition，HepAD38 cells transfected with shCREB1 plasmid（shCREB1 cells） and HepAD38 cells transfected with control plasmid with normal CREB1 expression（shControl cells） were divided into PBS control group and cisplatin treatment group，and the expression of HBV proteins and the level of HBV replication were measured. The results of qRT-PCR showed that compared with the cisplatin treat－ment group of shCREB1 cells，the cisplatin treatment group of shControl cells had significantly higher relative expression levels of HBV DNA（518.300±3.458 vs. 265.300±3.125，P=0.000） and HBV mRNA（2.713±0.318 vs. 1.263±0.056，P=0.000）. According to the results of Western blot，the cisplatin treatment group of shControl cells had a relative expression level of HBsAg of 1.254±0.001 and a relative expression level of HBcAg of 2.238±0.041，while the cisplatin treatment group of shCREB1 cells had a relative expression level of HBsAg of 1.121±0.036 and a relative expression level of HBcAg of 1.681±0.015；there were significant differ－ences in the relative expression levels of HBsAg and HBcAg between the two groups（P=0.021 and 0.000）. Conclusion：Cisplatin can increase the level of HBV replication and the expression levels of HBV proteins via CREB1.