Original Article

Effects of reactive oxygen species on lymphokine-activated killer cells in patients with bladder cancer.

Authors: Zhi-Ping WANG, Yi-Rong CHEN, Rong-Liang ZHENG, Jian-Zhong LU, Jia-Ji WANG

Abstract

AIM: To investigate the effect of reactive oxygen species on the proliferation of
lymphokine-activated killer (LAK) cells in patients with bladder cancer and their
cytolysis to bladder tumor cells.
METHODS: Sodium nitroprusside (SNP) was used as nitric oxide (NO) donor. The
superoxide anion (O2-.) was generated in the complete medium (CM) supplemented
with N-methylphenazonium methyl sulfate (PMS) 3-120 micromol/L and nicotinamide
adenine dinucleotide (NADH) 18 - 600 micromol/L. The hydroxyl radical (.OH) was
produced by adding ascorbic acid (AA) 0.5 - 400 micromol/L and ferrous sulfate
(Fe2+) 0.05 - 40 micromol/L in CM. LAK cell proliferation and cytotoxicity were
assayed in the presence of NO, .OH, or O2-. Bladder cancer cell lines BIU-87 and
EJ were cultured as target cells and cytotoxicity of LAK cells were determined by
MTT assay.
RESULTS: The proliferation of LAK cells induced by interleukin-2 (IL-2) was
inhibited by hydroxyl radical from 48 h to 96 h in a dose-dependent fashion and
was inhibited to 34.5 % compared with control at 96 h in the concentration of
ascorbic acid 400 micromol/L and ferrous sulfate 40 micromol/L. The inhibition
induced by.OH can be overcome by certain concentrations of mannitol or editic
acid. On the contrary, the proliferation of LAK cells induced by IL-2 was
stimulated by certain concentrations of NO or O2-. The stimulation induced by
O2-. can be overcome to control level by superoxide dismutase (SOD) 3 10(5) U/L.
Exogenous O2-. resulted in an increase in cytotoxicity of LAK cells against BIU87
and EJ cells. However, the LAK cells cytotoxicity treated with hydroxyl radical
or SOD showed no difference as compared with the control.
CONCLUSION: NO and O2-. enhanced the proliferation and activation and O2-.
up-regulated antitumor cytotoxicity of LAK cells in patients with bladder cancer.
The growth of LAK cells induced by IL-2 was down-regulated by hydroxyl radical.
The effects of these reactive oxygen species on the proliferation of LAK cells
induced by IL-2 were different.
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