[Establishment of a human laryngeal carcinoma Hep-2/5-Fu cell line and the screening of differentially expressed genes].

Objective:To establish a Hep-2/5-Fu of human drug-resistant laryngeal carcinoma cell line,and to screen the possible drug resistance-associated genes.Method:Hep-2/5-Fu of a human drug resistant laryngeal carcinoma cell line was induced by continuously exposing human laryngeal carcinom cells to gradually increasing concentrations of 5-Fu.The growth law was observed and the growth curve was protracted.The drug resistance of Hep-2/5-Fu was measured by MTT assay and the drug resistant index RI was calculated.Genes expressed differentially between Hep-2/5-Fu and its parent cell line Hep-2 were screened using a gene chip,and several selected drug resistance associated genes were confirmed by reverse transcription-polymerase chain reaction(RT-PCR).Result:Compared with its parental cells,the drug resistance cell line had slower growth rate and larger size.The Hep-2/5-Fu cell line showed cross drug resistance to 5-Fu,cisplatinand vincristine.There were 1210 differentially expressed genes possibly associated with drug resistance by the gene chip screening method.The possible drug resistance-related genes included Cyclin D,IGF-BP3,CASP9,and CDK4/6.The expression of Cyclin D in the Hep-2/5-Fu cell line was 6.599 7 times of that in the parent cell line.RT-PCR results were consistent with the gene chip results.Conclusion:The altered biological properties of Hep-2/5-Fu may be related to its drug resistance phenotype.Several genes,such as Cyclin D,are possibly involved in the mechanism of drug resistance in this cell line.