Clinical Investigation
Rosiglitazone Reverses Mitomycin C Resistance in Human Gastric Cancer Cells

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Abstract

Introduction

To explore the mechanisms of rosiglitazone (ROS), a selective peroxisome proliferator-activated receptor gamma ligand, in reversing mitomycin C (MMC) resistance in a human drug-resistant gastric cancer cell line.

Methods

The vincristine-resistant human gastric cancer cell line SGC7901/VCR and its parental cell line SGC7901 were treated with ROS, MMC (negative control), cyclosporine A+MMC (positive control) or ROS+MMC. A tetrazolium blue (methyl thiazolyl tetrazolium) assay was used to evaluate the sensitivity to these treatments. Flow cytometry analysis and acridine orange-ethidium bromide (AO-EB) fluorescent staining were used to determine the effects of ROS on MMC-induced apoptosis. Reverse transcription polymerase chain reaction and western blotting were used to measure the expression of multidrug resistant 1 (MDR1), Livin and P-glycoprotein (P-gp).

Results

ROS administration dose dependently increased the reversal index in MMC-treated SCG7901/VCR cells. ROS increased apoptosis in SGC7901/VCR cells compared with the blank group and MMC group. ROS+MMC also increased apoptosis in SGC7901/VCR cells compared with other groups (P < 0.05 or P < 0.01). The mRNA expression of MDR1 and Livin and the protein expression of P-gp in SGC7901/VCR cells were significantly higher than those in SGC7901 cells (P < 0.01). However, ROS or ROS+MMC treatment markedly upregulated the mRNA expression of MDR1 and Livin and the protein expression of P-gp in SGC7901/VCR cells (P < 0.01).

Conclusions

ROS reverses MMC resistance in human gastric cancer SGC7901/VCR cells by reducing expression of MDR1, Livin and P-gp and increasing apoptosis.

Section snippets

Materials

RPMI 1640 culture medium and fetal bovine serum (without Mycoplasma) was obtained from Gibco (Gibco BRL, New York, NY). VCR, methyl thiazolyl tetrazolium (MTT), trypsin and mitomycin C (MMC) were obtained from Sigma (Sigma, St. Louis, MO). The AMV first-strand cDNA synthesis kit, the ready-to-use reverse transcription polymerase chain reaction (RT-PCR) kit and primers for MDR1, including Livin and glyceraldehyde 3-phosphate dehydrogenase (GAPDH), were purchased from the Biological Engineering

ROS Reversed the Resistance of SGC7901/VCR to MMC as Determined by the MTT Assay

As shown in Figure 1, ROS reversed the resistance of SGC7901/VCR cells to MMC in a concentration-dependent manner. When the concentration of ROS increased progressively from 0 to 80 μM, the IC50 of MMC for SGC7901/ VCR cell line decreased from 9.13 to 0.87 mg/L. At 40 and 80 μM, ROS reversed the drug resistance of the SGC7901/ VCR cells, with RIs of MMC for SGC7901/VCR cells being 9.6 and 10.5, respectively, which were equal to the results with CSA (RI = 9.8, P > 0.05). These findings suggest

DISCUSSION

MDR is a significant problem in the treatment of gastric carcinomas. Many methods have been used to attempt to reverse MDR, including reversing agents, cytokines, monoclonal antibodies and various nucleic acid technologies. Reversing agents such as verapamil, cyclosporine and its derivative PSC-833, a non-immunosuppressive cyclosporine D analog and 2nd generation ABCBl/P-gp/MDRl specific reversal agent, can reverse MDR to reduce chemotherapeutic resistance to some degree. However, the severe

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    This work was supported by grants from the Foundation of Natural Science of Hunan Province (no. 08JJ5002), the Foundation of Science and Technology of Hunan Province (no. 2009SK3138) and the Scientific Research Foundation from Health Department of Hunan Province (no. 2007110).

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