增强的细胞凋亡介导姜黄素对EB病毒转化的淋巴母细胞系增殖的抑制作用。

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

作者信息

Ranjan D, Johnston T D, Reddy K S, Wu G, Bondada S, Chen C

机构信息

Department of Surgery, University of Kentucky, Lexington, Kentucky 40536, USA.

出版信息

J Surg Res. 1999 Nov;87(1):1-5. doi: 10.1006/jsre.1999.5719.

DOI:10.1006/jsre.1999.5719

PMID:10527697

Abstract

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 EBV was promoted by cyclosporin A (CyA) and that curcumin (Cur), a natural phenol with known antioxidant and antitumor properties, blocked EBV-induced B cell immortalization. In the following experiments we show that Cur inhibits the proliferation of EBV-transformed lymphoblastoid cell lines (LCL) via enhanced apoptosis.

METHODS

LCL were generated by infecting freshly isolated human B cells with EBV (B95-8) for 12 h and coculturing with predetermined optimal concentrations of CyA (500 ng/ml) for 4 weeks. LCL were then either frozen for future use or propagated for immediate experiments. These cells were then plated in 96-well plates with 20 microM Cur or 0.1% DMSO (vehicle control). The number of immortalized colonies/well, cell count, and (3)H uptake were used as an index of immortalization. To assess apoptosis rate LCL were cultured with 0.1% DMSO or Cur (20 microM) for 0, 18, and 42 h in culture flasks and then stained with MC540 and H33342, as markers for apoptosis, and analyzed by FACS.

RESULTS

A profound inhibition of proliferation was seen in the LCL with 20 microM 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 +/- 12,093 to 3750 +/- 1500/well (P = 0.002), and (3)H uptake reduced from 40,889 +/- 3669 to 70 +/- 5.2/well (P = 0.001). The apoptosis rate of LCL in the DMSO control at 24.07 and 16.87% increased significantly with 20 microM 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 appears to be mediated through enhanced apoptosis. A further investigation of this effect may be useful in prevention and therapy of B-cell lymphoma in immunodeficient patients.

摘要

背景

爱泼斯坦-巴尔病毒（EBV）相关的B细胞淋巴瘤在免疫缺陷状态下，如器官移植和HIV感染时更常见。我们之前报道过，环孢素A（CyA）可促进EBV介导的B细胞永生化，而姜黄素（Cur），一种具有已知抗氧化和抗肿瘤特性的天然酚类物质，可阻断EBV诱导的B细胞永生化。在接下来的实验中，我们表明Cur通过增强凋亡来抑制EBV转化的淋巴母细胞系（LCL）的增殖。

方法

通过用EBV（B95-8）感染新鲜分离的人B细胞12小时，并与预定的最佳浓度的CyA（500 ng/ml）共培养4周来生成LCL。然后将LCL冷冻以备将来使用或传代用于即时实验。然后将这些细胞接种到含有20 microM Cur或0.1% DMSO（溶剂对照）的96孔板中。永生化集落/孔的数量、细胞计数和³H摄取用作永生化的指标。为了评估凋亡率，将LCL在培养瓶中用0.1% DMSO或Cur（20 microM）培养0、18和42小时，然后用MC540和H33342染色，作为凋亡的标志物，并通过流式细胞术进行分析。

结果

与0.1% DMSO对照相比，20 microM姜黄素对LCL的增殖有显著抑制作用。集落计数从34.5±3.4降至0/孔（P = 0.005），细胞数量从101,250±12,093降至3750±1500/孔（P = 0.002），³H摄取从40,889±3669降至70±5.2/孔（P = 0.001）。DMSO对照中LCL的凋亡率在24.07%和16.87%，在20 microM Cur作用下，分别在18小时和42小时显著增加至76.4%和95.1%（P = 0.02）。