Enhanced apoptosis mediates inhibition of EBV-transformed lymphoblastoid cell line proliferation by curcumin

Background. Epstein-Barr virus (EBV)-associated B-cell lymphomas occur more frequently in immunodeficient states such as organ transplantation and HIV infection. We have previously reported that B cell immortalization with EB V was promoted by cyclosporin A (CyA) and that curcumin (Cur), a natural ph enol with known antioxidant and antitumor properties, blocked EBV-induced B cell immortalization. In the following experiments we show that Cur inhibi ts the proliferation of EBV-transformed lymphoblastoid cell lines (LCL) via enhanced apoptosis. Methods. LCL were generated by infecting freshly isolated human B cells wit h EBV (B95-8) for 12 h and coculturing with predetermined optimal concentra tions of CyA (500 ng/ml) for 4 weeks. LCL were then either frozen for futur e use or propagated for immediate experiments. These cells were then plated in 96-well plates with 20 mu M Cur or 0.1% DMSO (vehicle control). The num ber of immortalized colonies/well, cell count, and H-3 uptake were used as an index of immortalization. To assess apoptosis rate LCL were cultured wit h 0.1% DMSO or Cur (20 mu M) for 0, 18, and 42 h in culture flasks and then stained with MC540 and H33342, as markers for apoptosis, and analyzed by F AGS. Results. A profound inhibition of proliferation was seen in the LCL with 20 mu M curcumin compared to 0.1% DMSO control. The colony count reduced from 34.5 +/- 3.4 to 0/well (P = 0.005), cell number reduced from 101,250 +/- 1 2,093 to 3750 +/- 1500/well (P = 0.002), and 3H uptake reduced from 40,889 +/- 3669 to 70 +/- 5.2/well (P = 0.001). The apoptosis rate of LCL in the D MSO control at 24.07 and 16.87% increased significantly with 20 mu M Cur to 76.4 and 95.1% at 18 and 42 h, respectively (P = 0.02). Conclusion. Cur is a potent inhibitor of EBV-transformed LCL. This effect a ppears to be mediated through enhanced apoptosis. A further investigation o f this effect may be useful in prevention and therapy of B-cell. lymphoma i n immunodeficient patients. (C) 1999 Academic Press.