Establishment of a human laryngeal carcinoma Hep-2/5-Fu cell line and the screening of differentially expressed genes / 临床耳鼻咽喉头颈外科杂志

ABSTRACT

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 carcinoma 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, cisplatin and 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 6.5997 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.