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

Dinesh Ranjan, Thomas D. Johnston, Kunam Sudhakar Reddy, Guanghan Wu, Subbarao Bondada, Changguo Chen

Research output: Contribution to journalArticle

24 Citations (Scopus)

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 μM Cur or 0.1% DMSO (vehicle control). The number of immortalized colonies/well, cell count, and 3H uptake were used as an index of immortalization. To assess apoptosis rate LCL were cultured with 0.1% DMSO or Cur (20 μM) 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 μM 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 3H 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 μM 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.

Original languageEnglish (US)
Pages (from-to)1-5
Number of pages5
JournalJournal of Surgical Research
Volume87
Issue number1
DOIs
StatePublished - Nov 1999
Externally publishedYes

Fingerprint

Transformed Cell Line
Curcumin
Human Herpesvirus 4
Cell Proliferation
Apoptosis
Dimethyl Sulfoxide
Cell Line
B-Lymphocytes
B-Cell Lymphoma
Cyclosporine
Cell Count
Organ Transplantation
Phenol
HIV Infections
Antioxidants

Keywords

  • Apoptosis
  • B cell lymphoma
  • Curcumin
  • Lymphoblastoid cell line
  • PTLD

ASJC Scopus subject areas

  • Surgery

Cite this

Enhanced apoptosis mediates inhibition of EBV-transformed lymphoblastoid cell line proliferation by curcumin. / Ranjan, Dinesh; Johnston, Thomas D.; Reddy, Kunam Sudhakar; Wu, Guanghan; Bondada, Subbarao; Chen, Changguo.

In: Journal of Surgical Research, Vol. 87, No. 1, 11.1999, p. 1-5.

Research output: Contribution to journalArticle

Ranjan, Dinesh ; Johnston, Thomas D. ; Reddy, Kunam Sudhakar ; Wu, Guanghan ; Bondada, Subbarao ; Chen, Changguo. / Enhanced apoptosis mediates inhibition of EBV-transformed lymphoblastoid cell line proliferation by curcumin. In: Journal of Surgical Research. 1999 ; Vol. 87, No. 1. pp. 1-5.
@article{f0db6366160a4faeae1e1a9c722635ca,
title = "Enhanced apoptosis mediates inhibition of EBV-transformed lymphoblastoid cell line proliferation by curcumin",
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 μM Cur or 0.1{\%} DMSO (vehicle control). The number of immortalized colonies/well, cell count, and 3H uptake were used as an index of immortalization. To assess apoptosis rate LCL were cultured with 0.1{\%} DMSO or Cur (20 μM) 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 μM 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 3H 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 μM 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.",
keywords = "Apoptosis, B cell lymphoma, Curcumin, Lymphoblastoid cell line, PTLD",
author = "Dinesh Ranjan and Johnston, {Thomas D.} and Reddy, {Kunam Sudhakar} and Guanghan Wu and Subbarao Bondada and Changguo Chen",
year = "1999",
month = "11",
doi = "10.1006/jsre.1999.5719",
language = "English (US)",
volume = "87",
pages = "1--5",
journal = "Journal of Surgical Research",
issn = "0022-4804",
publisher = "Academic Press Inc.",
number = "1",

}

TY - JOUR

T1 - Enhanced apoptosis mediates inhibition of EBV-transformed lymphoblastoid cell line proliferation by curcumin

AU - Ranjan, Dinesh

AU - Johnston, Thomas D.

AU - Reddy, Kunam Sudhakar

AU - Wu, Guanghan

AU - Bondada, Subbarao

AU - Chen, Changguo

PY - 1999/11

Y1 - 1999/11

N2 - 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 μM Cur or 0.1% DMSO (vehicle control). The number of immortalized colonies/well, cell count, and 3H uptake were used as an index of immortalization. To assess apoptosis rate LCL were cultured with 0.1% DMSO or Cur (20 μM) 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 μM 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 3H 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 μM 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.

AB - 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 μM Cur or 0.1% DMSO (vehicle control). The number of immortalized colonies/well, cell count, and 3H uptake were used as an index of immortalization. To assess apoptosis rate LCL were cultured with 0.1% DMSO or Cur (20 μM) 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 μM 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 3H 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 μM 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.

KW - Apoptosis

KW - B cell lymphoma

KW - Curcumin

KW - Lymphoblastoid cell line

KW - PTLD

UR - http://www.scopus.com/inward/record.url?scp=0032719741&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0032719741&partnerID=8YFLogxK

U2 - 10.1006/jsre.1999.5719

DO - 10.1006/jsre.1999.5719

M3 - Article

C2 - 10527697

AN - SCOPUS:0032719741

VL - 87

SP - 1

EP - 5

JO - Journal of Surgical Research

JF - Journal of Surgical Research

SN - 0022-4804

IS - 1

ER -