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| Comparison of multi-drug resistant human hepatocellular carcinoma cell line Bel-7402/DOX model established by two methods |
| ZHONG Xingguo,GONG Renhua,SUN Dengqun,CAO Baoqiang,FAN Yulin,JIANG Shitao,CAI Jun,HE Xinmiao,LUO Huilai |
| Department of General Surgery, Anhui Provincial Corps Hospital,Chinese People’s Armed Police Forces, Hefei 230041, China |
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Abstract Objective To compare the biological characteristics of two types of human hepatocellular carcinoma multi-drug resistant cell sub-lines Bel-7402/DOX models established by two methods. Methods We established human hepatocellular carcinoma doxorubicin multi-drug resistant cell sub-lines models Bel-7402/DOXV and Bel-7402/DOXL by in vitro concentration gradient increased induction and nude mice liver-implanted induction, respectively. Phase contrast microscopy was used to observe the cells and MTT (methyl thiazolyl tetrazolium) method was used to detect drug resistance of the two different sub-lines of cells. The ingestion and excretion of cellular doxorubicin (DOX) and the expression of P-glycoprotein (P-gp), multi-drug resistance-sociated protein (MRP) and glutathione S-transfer enzyme system (GSH/GST) were detected by flow cytometry. Results The Bel-7402/DOXV and Bel-7402/DOXL generated cross-resistance to DOX and CDDP (cis-Diaminedichloroplatinum), they showed a significant difference in resistance to Bel-7402 IC50 value (P<0.01). The doubling times were significantly extended, compared with the parent cell line (39 h), and were 65 h (Bel-7402/DOXV) and 46 h (Bel-7402/DOXL). The excretion rates of DOX were significantly increased, compared with the parent cell (34.14%) line and were 81.06% (Bel-7402/DOXV) and 66.56% (Bel-7402/DOXL). Expressions of P-gp and MRP in the two groups of resistant sub-lines cells were significantly enhanced (P＜0.01). There was no significant variation in the expression of GSH/GST (P>0.05). Conclusions Stable resistance is involved in the resistant cell line model established by the two methods above. Liver implantation is a good simulation of human hepatocellular and proves to be an ideal model with characteristics similar to human hepatocellular biology and the pharmacokinetics of anticancer drugs.
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Received: 28 February 2014
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2014, Vol. 25 
