[Establishment of human multidrug-resistant hepatocellular carcinoma cell line (HepG2/Adm) and biological characteristics evaluation].
OBJECTIVE
Multidrug resistance (MDR) is considered to be the major obstacle for chemotherapy. In order to reverse tumor MDR in vitro, we designed this study to establish human multidrug-resistant hepatocellular carcinoma cell line (HepG2/Adm) and to investigate its biological characteristics.


METHODS
An adriamycin-resistant human hepatocellular carcinoma cell subline (HepG2/Adm) was established in vitro using gradually increased concentration of adriamycin (ADM) in culture. Cell growth was measured and multidrug resistance to multianticancer agents was evaluated by MTT assay. Flow cytometry (FCM) was performed to determine cell cycle,and to assess the expression of P-glycoprotein (P-gp), multidrug resistance-associated protein (MRP),lung-related protein (LRP),and glutathione S-transferase (GST), respectively. MDR mRNA expression related to these proteins were determined with reverse transcription polymerase chain reaction (RT-PCR).


RESULTS
Compared to HepG2, HepG2/Adm had a prolonged doubling time of 30.1 hours. The number of cells in S-phase was significantly decreased (5.6+/-0.03)% in HepG2/Adm while those in G1 and G2-phase increased [(4.2+/-0.09)% and (1.5+/-0.08)%, respectively]. Furthermore, HepG2/Adm was resistant to many anti-tumor agents, and its IC(50) of ADM was 26 times higher than that of parent cell line HepG2. Significant overexpression of P-gp, MRP, and GST were detected. RT-PCR showed that mRNA expression of P-gp, MRP, LRP, and GST were significantly increased in HepG2/Adm.


CONCLUSIONS
HepG2/Adm is human multidrug-resistant, and its resistance may be closely related to the overexpression of P-gp, MRP, and GST.
Zhai
Bao-Jin
BJ
The Center for Tumor Therapy, The Second Affiliated Hospital, Chongqing University of Medical Science, Chongqing, 400016, PR China. zhaibaojin@yahoo.com.cn
Wu
Feng
F
Shao
Ze-Yong
ZY
Hu
Kai
K
Wang
Zhi-Biao
ZB
D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1 / Q000032:analysis; D006528:Carcinoma, Hepatocellular / Q000188:drug therapy / Q000473:pathology*; D002453:Cell Cycle; D002455:Cell Division; D045744:Cell Line, Tumor; D002869:Chromosome Aberrations; D018432:Drug Resistance, Multiple; D019008:Drug Resistance, Neoplasm; D005434:Flow Cytometry; D006801:Humans; D008113:Liver Neoplasms / Q000188:drug therapy / Q000473:pathology*; D009363:Neoplasm Proteins / Q000032:analysis; D020394:Vault Ribonucleoprotein Particles; D000099187:Major Vault Protein
D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1; D009363:Neoplasm Proteins; D020394:Vault Ribonucleoprotein Particles; D000099187:Major Vault Protein