Establishment of the Clone of Gastrointestinal Stromal Tumor Cells with the Signs of Multiple Drug Resistance and Assessment of Its Properties

Abstract

The aim of the investigation was to obtain a tumor cell clone of gastrointestinal stromal tumors (GISTs) possessing resistance to chemotherapeutic agents of different mode of action, and to assess its sensitivity to various groups of chemotherapeutic agents.Materials and Methods. GIST cell line T-1 was used to obtain a clone of chemoresistant GIST cells. The viability assessment of tumor cells was carried out by using i-CELLigence RTCA cell analyzer (ACEA Biosciences, USA). The sensitivity of GIST T-1-29R cell clone to type II topoisomerase inhibitors (doxorubicin and etoposide); chemotherapeutic agents affecting the dynamic state of the spindle microtubules (vinblastine and paclitaxel); hydroxyurea; cisplatin; targeted drug imatinib were evaluated by using the MTT-based assay. Expression of apoptosis, the markers of DNA damage and chemoresistance was examined by immunoblotting.Results. A clone of GIST T-1 tumor cell line with the signs of chemoresistance (T-1-29R) was obtained after 8-month of cultivation of GIST tumor cells in the presence of the gradually increasing doses of paclitaxel. T-1-29R cells were found to possess resistance to paclitaxel, and cross-resistance to doxorubicin and etoposide, as well. Sensitivity of T-1-29R cells to other chemotherapeutic agents, including imatinib, did not change. Some multiple drug resistance proteins, e.g. MDR-1, were revealed to have an increased expression level in T-1-29R tumor cells when compared to parent T-1 cells.Conclusion. A clone of GIST T-1-29R cell line possesses phenotypical features of multiple drug resistance, that makes its perspective use for the assessment of GIST chemosensitivity, and for screening of new compounds for their cytotoxic and antitumor activities in vitro and in vivo, as well.

2-APCAs, the Novel Microtubule Targeting Agents Active against Distinct Cancer Cell Lines

“…The IM-resistant GIST T-1R subline was established in our laboratory after a continuous induction from 0.4 to 1000 nM IM in a stepwise increasing concentration manner [ 55 ]. PTX-resistant HCC1806-Tх-R and GIST T-1-Тx-R sublines were established in our laboratory after a continuous induction from 0.001 to 300 nM PTX in a stepwise increasing concentration manner [ 56 , 57 ]. All cell lines were maintained in Dulbecco’s modified Eagle’s medium or RPMI-1640 medium, supplemented with 10–15% fetal bovine serum, 1% L-glutamine, 50 U/mL penicillin, and 50 μg/mL streptomycin, and cultured in a humidified atmosphere of 5% CO2 at 37 °C (LamSystems, Myass, Russia).…”

Infigratinib (BGJ 398), a Pan-FGFR Inhibitor, Targets P-Glycoprotein and Increases Chemotherapeutic-Induced Mortality of Multidrug-Resistant Tumor Cells

“…GIST T-1 was established from a metastatic pleural tumor from a stomach GIST and contained a heterozygous 57-base pair deletion (V570-Y578) in the KIT exon 11 [44]. PTX-resistant ABCB1-overexpressing GIST T-1R subline was also established in our laboratory via stepwise treatment with increasing concentrations of PTX, as shown elsewhere [43]. The cell lines indicated above were maintained in RPMI-1640 medium (Paneco, Moscow, Russia) supplemented with 15% fetal bovine serum (Gibco; Thermo Fisher Scientific, Inc., Waltham, MA, USA), 50 U/mL penicillin and 50 µg/mL streptomycin.…”

“…Given that a broad spectrum of tumorous tissues acquires MDR phenotype due to the repeated exposure to chemotherapeutic agents, we also expanded our study to the cancer cell lines that originated from the epithelium of the gastrointestinal tract and bone tissues. Thus, we also included in our study ABCB1-overexpressing gastrointestinal stromal tumor (GIST T-1 Tx-R) cell subline, which was also generated in our laboratory, as shown elsewhere [43].…”