Heat-induced membrane damage combined with adriamycin on prostate carcinoma PC-3 cells: correlation of cytotoxicity, permeability and P-glycoprotein or metallothionein expression.
OBJECTIVE
To assess heat-induced membrane damage in a prostate cancer cell line when combined with adriamycin treatment.


METHODS
The changes in intracellular adriamycin accumulation, cell proliferation and cell-cycle fractions were examined after human prostate carcinoma PC-3 cells were exposed to heat and/or further treatment with adriamycin. Proliferation and the cell cycle were determined using adherent cell analysis and sorting laser cytometry (ACAS) or flow cytometry. P-glycoprotein (PGP) and metallothionein (MT) expression, which may have a physiological role in the transport of or reduction in cytotoxicity of some anticancer drugs, were also analysed after cells were exposed to heat, using immunohistochemical or flow cytometric methods.


RESULTS
There was a significant increase in intracellular adriamycin accumulation, related to both influx (P<0.05) and efflux (P<0.01), in cells treated with adriamycin, especially after heating them at 44 degrees C for 1 h. There was a significant decrease in cell proliferation of preheated cells when exposed to adriamycin, especially at 44 degrees C (P<0.05). In the cell-cycle analysis, cells preheated at 44 degrees C showed partial accumulation in the debris or apoptotic fraction at 24 h, and many cells accumulated in these fractions at 48 h. There was significantly less PGP or MT expression in cells preheated at 44 degrees C than in control cells or cells preheated at 41 degrees C (P<0.01). This reduction in PGP or MT level by heating may inhibit drug efflux and thus increase intracellular drug level at elevated temperatures.


CONCLUSIONS
These results suggest that hyperthermia may damage the drug-exclusion mechanism in these cells and thus increase the effectiveness of drug action.
Moriyama-Gonda
N
N
Department of Urology, Shimane Medical University, Izumo, Japan.
Igawa
M
M
Shiina
H
H
Wada
Y
Y
D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1 / Q000378:metabolism; D000970:Antineoplastic Agents / Q000378:metabolism / Q000627:therapeutic use*; D002455:Cell Division; D002462:Cell Membrane; D002463:Cell Membrane Permeability; D004273:DNA, Neoplasm / Q000032:analysis; D004317:Doxorubicin / Q000378:metabolism / Q000627:therapeutic use*; D005434:Flow Cytometry; D006358:Hot Temperature / Q000627:therapeutic use*; D006801:Humans; D007150:Immunohistochemistry; D008297:Male; D008668:Metallothionein / Q000378:metabolism; D009363:Neoplasm Proteins / Q000378:metabolism; D011471:Prostatic Neoplasms / Q000188:drug therapy* / Q000378:metabolism / Q000473:pathology
D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1; D000970:Antineoplastic Agents; D004273:DNA, Neoplasm; D009363:Neoplasm Proteins; D004317:Doxorubicin; D008668:Metallothionein