Establishment of a GIST-T1 gastrointestinal stromal tumour cell line resistant to imatinib mesylate

Abstract

In the present study, imatinib mesylate (IM) was used to induce resistance in the gastrointestinal stromal tumour (GIST) cell line, GIST-T1, to establish a stable resistant cell line. The growth characteristics and expression profile of the established cell line were compared with those of the parental cell line. Additionally, the resistance mechanism of the gastrointestinal stromal tumours was preliminarily investigated. The GIST-T1 cells were cultured in vitro, and the drug was administered in the logarithmic phase of cell growth using intermittent dosing with increasing concentrations to obtain a drug-resistant cell line by repeated induction. Differences in the biological behaviours of the parental cells and drug-resistant cells were examined, and changes in the expression profiles were compared in the two cell lines. The results showed that the IM-resistant GIST-T1 cell line (GIST-T1 IR) was successfully established. Analysis of the biological behaviours of the two cell lines revealed that the average doubling times of the parental cells and drug-resistant cells were 26.59 and 33.63 h, respectively. The results of a scratch migration assay revealed that the migration ability was enhanced in the GIST-T1 IR cells. The results of CCK-8 detection indicated that the half maximal inhibitory concentration values of the two types of cells were 10.5 and 42.0 µM, respectively, which represented an increase of ~4-fold in the GIST-T1 IR cells. Flow cytometric cell cycle analysis indicated that the numbers of cells in the G0/G1, S and G2 phases increased following the induction treatment. Taken together, an IM-resistant GIST T1 cell line was successfully established, which opens novel avenues for individualized tumour chemotherapy.

HIF-1α regulates cellular metabolism, and Imatinib resistance by targeting phosphogluconate dehydrogenase in gastrointestinal stromal tumors

“…IM-resistant cell lines, GIST-T1R and GIST-882R, were obtained from sensitive parental GIST-T1 and GIST-882. To establish IM resistant cell lines, drug-containing medium with intermittently increasing IM (Glivec, Novartis AG, Switzerland) concentrations (from 5 μM and gradually increasing to 50 μM) was added to culture medium when GIST cells in the exponential phase 46 . After culturing for 48 h, the IM-containing medium was replaced.…”

SPK1/S1P axis confers gastrointestinal stromal tumors (GISTs) resistance of imatinib

“…For cellular morphological observation, the GIST-T1 and GIST-T1/IMR cells were displayed in the medium for the morphological observation under a light microscope from 10 to 40 amount of magnification. As indicated by previous report [ 32 ], cell growth curve and calculation of resistance index were carried out. Afterwards, to verify if SPK1 inhibitor N , N -dimethylsphingosine (DMS), could confer the sensitivity of IM treatment, these two cells were introduced onto the bottom of the 6-well culture plate with 5 × 10 5 /mL cells per well for cell scratch assay.…”