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Pit cells (hepatic natural killer cells) of the rat induce apoptosis in colon carcinoma cells by the perforin/granzyme pathway
Author(s) -
Vermijlen David,
Luo Dianzhong,
Robaye Bernard,
Seynaeve Carine,
Baekeland Marijke,
Wisse Eddie
Publication year - 1999
Publication title -
hepatology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 5.488
H-Index - 361
eISSN - 1527-3350
pISSN - 0270-9139
DOI - 10.1002/hep.510290143
Subject(s) - perforin , cytotoxic t cell , apoptosis , granzyme b , granzyme , biology , programmed cell death , fas ligand , cancer cell , interleukin 21 , cell culture , microbiology and biotechnology , lymphokine activated killer cell , dna fragmentation , cancer research , in vitro , biochemistry , cancer , genetics
The high mortality of colon cancer is to a large extent caused by the frequent occurrence of liver metastasis. This is remarkable, because the liver harbors two distinct cell populations that can eliminate invading cancer cells, namely hepatic natural killer (NK) cells and Kupffer cells. These hepatic NK cells, known as pit cells, are the most cytotoxic cells of the naturally occurring NK cells. However, the mechanism by which pit cells eliminate tumor cells is largely unknown. Because we recently found an indication that apoptosis is involved, we tried to assess the role of this mode of cell death using an in vitro system with isolated pure pit cells (>90%) and CC531s cells, a rat colon carcinoma (CC) cell line. Pit cells induced apoptosis in CC531s cells as shown by quantitative DNA fragmentation, agarose gel electrophoresis, and different modes of microscopy. When extracellular Ca 2+ was chelated by ethylene glycol‐bis(β‐aminoethyl ether)‐ N , N ,‐tetraacetic acid (EGTA) during coincubation or when the pit cells were preincubated with the granzyme inhibitor 3,4‐dichloroisocoumarin (DCI), the induction of apoptosis was abolished. These results show that pit cells use the Ca 2+ ‐dependent perforin/granzyme pathway to induce apoptosis in the CC531s cells, and not the alternative Ca 2+ ‐independent Fas pathway. To further exclude the possibility of the involvement of the Fas pathway, we treated CC531s cells with recombinant Fas ligand. This treatment did not result in the induction of apoptosis, indicating that CC531s cells are resistant to Fas‐mediated apoptosis. We conclude therefore that pit cells induce apoptosis in CC cells in vitro by the perforin/granzyme pathway.