Abstract
All-trans retinoic acid (RA) was previously shown to regulate the growth of gastric cancer cells derived from the cell line SC-M1. This study was designed to investigate the effect of RA on the sensitivity of SC-M1 cells to lymphokine-activated killer (LAK) activity. RA at the concentration range of 0.001-10 microM was shown to induce SC-M1 cells to exhibit resistance to LAK activity in a dose-dependent manner. A kinetics study indicated that a significantly increased resistance was detected after 2 days of co-culturing SC-M1 cells with RA and reached a maximum after 6 days of culture. Similar results were obtained from two other cancer cell lines: promyelocytic leukaemia HL-60 and hepatic cancer Hep 3B. A binding assay demonstrated that the binding efficacy between target SC-M1 cells and effector LAK cells was not altered by RA. Flow cytometric analyses revealed that RA exhibited no effect on the expression of cell surface molecules, including HLA class I and class II antigens, intercellular adhesion molecule-1 and -2, and lymphocyte function antigen-3. Cell cycle analysis revealed that culture of SC-M1 cells with RA resulted in an increase in G0/G1 phase and a decrease in S phase, accompanied by a decrease in cyclin A and cyclin B1 mRNA as determined by Northern blot analysis. Additionally, RA was shown to enhance the expression of retinoic acid receptor alpha (RAR alpha) in SC-M1 cells, and to have no effect on the expression of RARbeta or RARgamma. Taken together, these results indicate that RA can significantly increase gastric cancer cells SC-M1 to resist LAK cytotoxicity by means of a cytostatic effect through a mechanism relating to cell cycle regulation. The prevailing ideas, such as a decrease in effector to target cell binding, a reduced MHC class I antigen expression or an altered RARbeta expression, are not involved.
Highlights
SC-Mi cells with retinoic acid (RA) resulted in an increase in GJG, phase and a decrease in S phase, accompanied by a decrease in cyclin A and cyclin Bi
SC-Mi cells with RA resulted in an increase in GJG, phase and a decrease in S phase, accompanied by a decrease in cyclin A and cyclin Bi mRNA as determined by Northern blot analysis
RA was shown to enhance the expression of retinoic acid receptor a (RARa) in SC-Mi cells, and to have no effect on the expression of RARfi or RARy
Summary
Target cellsThree cancer cell lines, including gastric cancer SC-MI, promyelocytic leukaemia HL-60 and hepatic cancer Hep 3B, were used as target cells in the cytotoxicity assay. Tumour cells were maintained in RPMI-1640 medium with 10% fetal calf serum (FCS) (Gibco, Grand Island, NY, USA). Peripheral blood mononuclear cells (PBMCs) were obtained from normal healthy volunteers and incubated at 37°C in complete medium containing 3000 IU ml-' IL-2, under a moist atmosphere with 5% carbon dioxide in culture flasks at a cell concentration of 2-3 x 106 ml'. Complete medium contained RPMI-1640 with 10% FCS, 0.3 mg ml-1 L-glutamine, 100 gg ml-' streptomycin and 100 U ml-' penicillin. Activated killer cells were harvested after culture for 4 days and washed twice with RPMI- 1640. The viable cells were counted and resuspended in RPMI-1640 containing 10% FCS for the standard 4-h 5'Cr-release assay (Chao et al, 1990, 1995a), and for surface marker studies with flow cytometry (Chao et al, 1995b)
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