MDR1 gene expression: its effect on drug resistance to doxorubicin in human hepatocellular carcinoma cell lines.
BACKGROUND
Hepatic tumors are resistant to many chemotherapeutic agents. Although elevated MDR1 (also known as PGY1) gene expression has been shown in such tumors, no direct association has been established between the gene expression and multidrug resistance.


OBJECTIVE
To evaluate the role of the MDR1 gene in the drug resistance of hepatoma, we tested nine human hepatoma cell lines for their expression of the MDR1 gene.


METHODS
We measured the MDR1 messenger RNA (mRNA) expression by RNA slot-blot analysis and by immunocytochemical staining with a P-glycoprotein-specific monoclonal antibody, MRK16. The in vitro chemosensitivity of these cell lines to fluorouracil, doxorubicin, mitomycin C, cisplatin, and etoposide (VP-16) was determined using the MTT (3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide) colorimetric assay. For doxorubicin cytotoxicity, we also tested the potentiating effect of several multidrug resistance-reversing agents.


RESULTS
Slot-blot analysis and immunocytochemistry showed that two cell lines expressed high levels of MDR1 mRNA, one expressed an intermediate level, and all others were low expressors. The MTT assay results showed that all cell lines tested were generally resistant to chemotherapeutic agents. The assay area under the curve (AUC) was within a clinically achievable range only for VP-16 in one of nine cell lines. When the IC50 values were compared among the cell lines, the results revealed a close association with the MDR1 gene expression only for doxorubicin resistance. Verapamil and quinidine lowered the IC50 values of doxorubicin for MDR1-positive cell lines. The lowered assay AUC levels for both reversing agents, however, were still higher than the clinically achievable range.


CONCLUSIONS
These results indicate that the MDR1 gene probably has a role in doxorubicin resistance in hepatocellular carcinoma and that the resistance can be overcome by some multidrug resistance-reversing agents.


CONCLUSIONS
Some widely used anticancer agents might be ineffective for treating hepatocellular carcinoma in clinical situations even when combined with reversing agents.
Park
J G
JG
Laboratory of Cell Biology, Seoul National University College of Medicine, Korea.
Lee
S K
SK
Hong
I G
IG
Kim
H S
HS
Lim
K H
KH
Choe
K J
KJ
Kim
W H
WH
Kim
Y I
YI
Tsuruo
T
T
Gottesman
M M
MM
D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1; D006528:Carcinoma, Hepatocellular / Q000188:drug therapy / Q000378:metabolism*; D002352:Carrier Proteins / Q000037:antagonists & inhibitors*; D002438:Cefoperazone / Q000494:pharmacology; D002443:Ceftriaxone / Q000494:pharmacology; D004317:Doxorubicin / Q000494:pharmacology*; D004351:Drug Resistance / Q000235:genetics*; D004357:Drug Synergism; D015870:Gene Expression / Q000187:drug effects; D006801:Humans; D007150:Immunohistochemistry; D008113:Liver Neoplasms / Q000188:drug therapy / Q000378:metabolism*; D008562:Membrane Glycoproteins / Q000037:antagonists & inhibitors*; D009363:Neoplasm Proteins / Q000037:antagonists & inhibitors*; D011802:Quinidine / Q000494:pharmacology; D012333:RNA, Messenger / Q000032:analysis; D014268:Trifluoperazine / Q000494:pharmacology; D014407:Tumor Cells, Cultured; D014700:Verapamil / Q000494:pharmacology
D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1; D002352:Carrier Proteins; D008562:Membrane Glycoproteins; D009363:Neoplasm Proteins; D012333:RNA, Messenger; D014268:Trifluoperazine; D002443:Ceftriaxone; D002438:Cefoperazone; D004317:Doxorubicin; D014700:Verapamil; D011802:Quinidine