Abstract
The objective of the present study was to investigate the multicellular resistance of human hepatocarcinoma cells BEL-7402 to pharmorubicin. Cells (1 x 10(4)) and 200 microcarrier Cytodex-3 beads were seeded onto a 24-well plate and cultured in RPMI 1640 medium. After the formation of multicellular aggregates, morphology and cell viability were analyzed by scanning electron microscopy, transmission electron microscopy and flow cytometry, respectively. The IC50 was determined by flow cytometry and MTT assay after the cells cultured in aggregates and monolayers were treated with pharmorubicin. The culture products exhibited structural characteristics somewhat similar to those of trabecular hepatocarcinoma in vivo. Among the microcarriers, cells were organized into several layers. Intercellular spaces were 0.5-2.0 microm wide and filled with many microvilli. The percent of viable cells was 87%. The cells cultured as multicellular aggregates were resistant to pharmorubicin with IC50 4.5-fold and 7.7-fold that of monolayer culture as determined by flow cytometry and MTT assay, respectively. This three-dimensional culture model may be used to investigate the mechanisms of multicellular drug resistance of hepatocarcinoma and to screen new anticancer drugs.
Highlights
Intrinsic or acquired resistance to chemotherapeutic drugs is one of the major obstacles in the treatment of solid tumors
Human hepatocarcinoma cells BEL-7402 were cultured with Cytodex-3 microcarrier beads to form 3-D aggregates
The results indicated that the cells were oval spheroid or polyhedric
Summary
Intrinsic or acquired resistance to chemotherapeutic drugs is one of the major obstacles in the treatment of solid tumors. A number of possible mechanisms have been proposed to account for drug resistance [1]. Previous research demonstrated that tumor cells cultured in three-dimensional (3-D) aggregates were resistant to cytotoxic drugs. The resistance was lost when the cells were disaggregated and cultured in monolayers. Resistance could be acquired again when cells were cultured in 3-D aggregates [2,3]. Many kinds of tumor cells become resistant to cytotoxic drugs when cultured in 3-D aggregates. Three-dimensional culture makes it possible to directly relate structure to function and to reveal the response of cells to chemotherapeutic drugs similar to that observed in vivo. Three-dimensional culture of hepatocarcinoma cells has been reported [4,5,6,7], but hepatocarcinoma cells with different characteristics need to be investigated
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Schedule a callMore From: Brazilian journal of medical and biological research = Revista brasileira de pesquisas medicas e biologicas
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