To establish a cytologic expressing system of rat glutathione S-transferase pi (GST-pi) cDNA for detecting the resistance of HeLa cells to anticancer drugs. Methods The assessment was made with various anticancer drugs (adriamycin, mitomycin, cisplatinum and vincristine) that showed different cytotoxicities in transfectant HeLa cells with pSV-GT containing rat GST-pi cDNA (HeLa/pSV-GT) or control pSV-neo (HeLa/pSV-neo). Expression levels of GST-pi mRNA in HeLa/pSV-GT and HeLa/pSV-neo were measured by in situ hybridization using Digoxin-labelled cDNA probe. Results HeLa/pSV-GT expressed significantly high degree of GST-pi mRNA, whereas both HeLa/pSV-neo and HeLa cells had very low expression. Cytotoxicities of HeLa/pSV-GT and HeLa/pSV-neo with 4 anticancer drugs were measured by MTT assay. Drug concentrations for yielding 50% inhibition (IC50) in HeLa/pSV-GT by adriamycin, mitomycin and cisplatinum were 70.13 靏/mL, 10.95 靏/mL and 16.52 靏/mL, respectively. In contrast, IC50 in HeLa/pSV-neo was 10.34 靏/mL, 7.48 靏/mL and 13.70 靏/mL, respectively. The cytotoxicities of vincristine on both HeLa/pSV-GT and HeLa/pSV-neo were not significantly different. Conclusions Our findings suggest that HeLa/pSV-GT containing rat GST-pi cDNA is resistant to some anticancer drugs due to overexpression of GST-pi. Also, HeLa/pSV-GT cell line could serve as a useful cytogenetic model for further research. To establish a cytologic expressing system of rat glutathione S-transferase pi (GST-pi) cDNA for detecting the resistance of HeLa cells to anticancer drugs. Methods The assessment was made with various anticancer drugs (adriamycin, mitomycin, cisplatinum and vincristine) different cytotoxicities in transfectant HeLa cells with pSV-GT containing rat GST-pi cDNA (HeLa / pSV-GT) or control pSV-neo (HeLa / pSV- neo). Expression levels of GST-pi mRNA in HeLa / pSV-GT and HeLa / pSV-neo were measured by in situ hybridization using Digoxin-labeled cDNA probe. Results HeLa / pSV-GT expressed significantly high degree of GST-pi mRNA, while both HeLa / pSV-neo and HeLa cells had very low expression. Cytotoxicities of HeLa / pSV-GT and HeLa / pSV-neo with 4 anticancer drugs were measured by MTT assay. Drug concentrations for yielding 50% inhibition (IC50) in HeLa / pSV-GT by adriamycin, mitomycin and cisplatinum were 70.13 靏 / mL, 10.95 靏 / mL and 16.52 靏 / mL, respectively. In contrast, IC50 in The cytotoxicities of vincristine on both HeLa / pSV-GT and HeLa / pSV-neo were not significantly different. Conclusions Our findings suggest that that HeLa / pSV-neo was 10.34 靏 / mL, 7.48 靏 / mL and 13.70 靏 / mL, respectively. HeLa / pSV-GT containing rat GST-pi cDNA is resistant to some anticancer drugs due to overexpression of GST-pi. Also, HeLa / pSV-GT cell line could serve as a useful cytogenetic model for further research.