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. 2018 May;15(5):7589-7594.
doi: 10.3892/ol.2018.8283. Epub 2018 Mar 16.

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

Yongjian Zhou  1 , Jiabi Chen  2 , Xiaoyuan Weng  1 , Guosheng Lin  1 , Zicheng Huang  3 , Hanli Shui  1
Affiliations

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

Yongjian Zhou et al. Oncol Lett. 2018 May.

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 VSports手机版. Taken together, an IM-resistant GIST T1 cell line was successfully established, which opens novel avenues for individualized tumour chemotherapy. .

Keywords: GIST-T1; cell line; gastrointestinal stromal tumour; imatinib mesylate; resistance mechanism V体育安卓版. .

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Figures

Figure 1.
Figure 1.
GIST-T1 and GIST-T1 IR cells. (A) GIST-T1 cell density was adjusted to 5, 000 cells/100 µl, and the cells were cultured in 2.5, 10, 15, 20, 25, 30, 40 and 50 µM IM. (B) GIST-TI IR cell medium contained 5, 25, 30, 35, 40, 50, 62.5 and 75 µM IM, and the cells were cultured at the same cell density. GIST, gastrointestinal stromal tumour; IM, imatinib mesylate; IR, IM-resistant.
Figure 2.
Figure 2.
GIST-T1 and GIST-T1 IR hematoxylin and eosin staining (×400 magnification). (A) GIST-T1 cells were elongated, spindle-shaped, mononuclear or binuclear, and had a relatively large volume. (B) GIST-T1 IR cells had a smaller volume, darkly stained nuclei, were predominantly binuclear, triple-nuclear or multiple-nuclear, and had large nuclei. IR, imatinib mesylate-resistant; IC50, half maximal inhibitory concentration.
Figure 3.
Figure 3.
GIST-T1 and GIST-T1 IR cell growth curves. The GIST-T1 and GIST-T1 IR cells were seeded into 24-well plates at a concentration of 5×104 cells/ml and counted every 24 h for 5 days consecutively. IR, imatinib mesylate-resistant.
Figure 4.
Figure 4.
GIST-T1 and GIST-T1 IR cell cycle. The GIST-T1 and GIST-T1 IR cell cycles were measured by flow cytometry. IR, imatinib mesylate-resistant.
Figure 5.
Figure 5.
GIST-T1 and GIST-T1 IR cell scratch assay. The GIST-T1 and GIST-T1 IR cells were treated with IM at a concentration of 0 (−) or 25 (+) µM. Migration was observed following incubation for 24 h. IM, imatinib mesylate; IR, IM-resistant; T1, non-resistant.
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