Establishment And Characterization Of Tongual Chemoresistant Squamous Cell Carcinoma Cell Strain

Establishment and characterization oftongual chemoresistantsquamous cell carcinoma cell strainObjective:The chemoresistant cell lines are established by expose tongual squamous cell carcinoma parent cell line to chemotherapeutical drug. Compare their characterization and gene expression profile to the parent cell line. make a primary reseaching of the mechanisms of drug resistance of head and neck squamous cell Carcinoma in order to provid of basis chemotherapy.Methods:l. The cell line of tongual squamous cell carcinoma Tca8113 is cultured in vitro. The gradient acute toxitical experiment of cell line Tca8113 is took on by MTT array in order to research the effect of different concentration of chemotherapeutical drug to the tongual squamous cell carcinoma cell line Tca8113, to look for adaptive initial drug concentration and upgrad extent of screening and induction of chemoresistant cell line;2. Exposing cell line Tca8113 to carboplatin or 5- flurouracil periodically to kill all most of sensitive cell, then the remnant cells are cultured in drug-free medium to recover their normal proliferation and passage. Repeating the course, finally, two chemoresistant cell are established and are named Tca8113/Ca and Tca8113/Fu respectively.3. Comparing cell morpholopy, population doubling time and cell cycle distribution of Tca8113/Ca and Tca8113/Fu to parent cell line Tca8113 respectively;4. Isolating totalRNA from Tca8113/Ca, Tca8113/Fu and parent cell line Tca8113, then synthesis double-stranded cDNA, biotin-labeled cRNA, oligonucleotide microarray prepare, microarray hybridization, Washing, Staining, and Scanning, finally, data analyselResults:1. Two chemoresistant cell lines to carboplatin or 5- flurouracil are established from parent cell line Tca8113 by exposed it to carboplatin or 5- flurouracil periodically and are named Tca8113/Ca (RI:2.86) and Tca8113/Fu (RI:2.31);2. Comparing to parent cell line Tca8113, morphology changes, doubling period longers and cell cycle distributing changes in Tca8113/Ca; morphologychanged in Tca8113/Fu, but not doubing period and cell cycle distributing;3. In global human genewhich be detected by genechip, 67 genes displays different expression of Tca8113/Ca:21gene are up-regulated, 46 are down-regulated; 33 genes displays different expression of Tca8113/Fu:14genes are up-regulated and 19 are down-regulated;Conclusion:1. Two chemoresistant line of tongual squamous cell carcinoma Tca81113/Ca to carboplatin or Tca8113/Fu to 5- flurouracil are establelished successfully;2. The gene expression profile are compared by global gene microarray, some gene cluster about DNA-dependent regulation of transcription and transcription display different expression both two drug resistant cell line. we hypothesize that these gene cluster may have something to with chemoresistance, the cell motility related'to genes of gene expressing profiling of Tca8113/Ca change.