Sapitinib Reverses Anticancer Drug Resistance in Colon Cancer Cells Overexpressing the ABCB1 Transporter

Gao, Hailing; Gupta, Pranav; Cui, Qingbin; Ashar, Yunali V.; Wu, Zhuo‐Xun; Zeng, Leli; Lei, Zi‐Ning; Teng, Quincy; Ashby, Charles R.; Guan, Yingjun; Chen, Zhe‐Sheng

doi:10.3389/fonc.2020.574861

Cited by 21 publications

(14 citation statements)

References 59 publications

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“…The elevated sensitivity to paclitaxel from MCTSs generated from SW620/Ad300‐ABCB1ko and SW620‐ABCB1ko cells were shown by loss of integrity and loosely associated dead cells at the outer layer after 48 and 72 h of exposure, which was consistent with the appearance of paclitaxel‐treated MCTSs from SW620/Ad300 cells reported by Gao et al. 37 …”

Section: Discussionsupporting

confidence: 87%

“…Cells were prepared for antibody binding by undergoing fixation, permeabilization, and serum blocking processes as previously described. 37 Cells were then incubated with the monoclonal antibody against ABCB1 (1:1000 dilution) overnight at 4°C followed by incubation with Alexa Flour 488‐linked secondary antibody (1:1000 dilution) for 2 h at room temperature. The nuclei of cells were visualized by counterstaining using DAPI.…”

Section: Methodsmentioning

confidence: 99%

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Overcoming multidrug resistance by knockout of ABCB1 gene using CRISPR/Cas9 system in SW620/Ad300 colorectal cancer cells

Lei

Teng

et al. 2021

MedComm

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Multidrug resistance (MDR) has been extensively reported in colorectal cancer patients, which remains a major cause of chemotherapy failure. One of the critical mechanisms of MDR in colorectal cancer is the reduced intracellular drug level led by the upregulated expression of the ATP‐binding cassette (ABC) transporters, particularly, ABCB1/P‐gp. In this study, the CRISPR/Cas9 system was utilized to target ABCB1 in MDR colorectal cancer SW620/Ad300 cell line with ABCB1 overexpression. The results showed that stable knockout of ABCB1 gene by the CRISPR/Cas9 system was achieved in the MDR cancer cells. Reversal of MDR against ABCB1 chemotherapeutic drugs increased intracellular accumulation of [ 3 H]‐paclitaxel accumulation, and decreased drug efflux activity was observed in MDR SW620/Ad300 cells after ABCB1 gene knockout. Further tests using the 3D multicellular tumor spheroid model suggested that deficiency in ABCB1 restrained tumor spheroid growth and restore sensitivity to paclitaxel in MDR tumor spheroids. Overall, the CRISPR/Cas9 system targeting the ABCB1 gene can be an effective approach to overcome ABCB1‐mediated MDR in colorectal cancer SW620/Ad300 cells.

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Section: Discussionsupporting

confidence: 87%

Section: Methodsmentioning

confidence: 99%

Overcoming multidrug resistance by knockout of ABCB1 gene using CRISPR/Cas9 system in SW620/Ad300 colorectal cancer cells

Lei

Teng

et al. 2021

MedComm

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“…Notably, four out of the six FDA-approved quinazolines are EGFR inhibitors: gefitinib, erlotinib, afatinib, and lapatinib [ 33 , 34 ]. It was also reported that the EGFR and HER2/3 inhibitor sapitinib, which contains a quinazoline moiety, showed chemosensitizing effect on resistant CRC cells by downregulation of the ABCB1 transporter [ 35 ]. A previously synthetized novel quinazoline derivative [ 36 ] will be used in this study (named HAA 2020 , Figure 1 C).…”

Section: Introductionmentioning

confidence: 99%

Chemosensitization of HT29 and HT29-5FU Cell Lines by a Combination of a Multi-Tyrosine Kinase Inhibitor and 5FU Downregulates ABCC1 and Inhibits PIK3CA in Light of Their Importance in Saudi Colorectal Cancer

et al. 2021

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Colorectal cancer (CRC) remains one of the main causes of death worldwide and in Saudi Arabia. The toxicity and the development of resistance against 5 fluorouracil 5FU pose increasing therapeutic difficulties, which necessitates the development of personalized drugs and drug combinations. Objectives: First, to determine the most important kinases and kinase pathways, and the amount of ABC transporters and KRAS in samples taken from Saudi CRC patients. Second, to investigate the chemosensitizing effect of LY294002 and HAA2020 and their combinations with 5FU on HT29, HT29-5FU, HCT116, and HCT116-5FU CRC cells, their effect on the three ABC transporters, cell cycle, and apoptosis, in light of the important kinase pathways resulting from the first part of this study. Methods: The PamChip® peptide micro-array profiling was used to determine the level of kinase and targets in the Saudi CRC samples. Next, RT-PCR, MTT cytotoxicity, Western blotting, perturbation of cell cycle, annexin V, and immunofluorescence assays were used to investigate the effect on CRC, MRC5, and HUVEC cells. Results: The kinase activity profiling highlighted the importance of the PI3K/AKT, MAPK, and the growth factors pathways in the Saudi CRC samples. PIK3CA was the most overexpressed, and it was associated with increased level of mutated KRAS and the three ABC transporters, especially ABCC1 in the Saudi samples. Next, combining HAA2020 with 5FU exhibited the best synergistic and resistance-reversal effect in the four CRC cells, and the highest selectivity indices compared to MRC5 and HUVEC normal cells. Additionally, HAA2020 with 5FU exerted significant inhibition of ABCC1 in the four CRC cells, and inhibition of PIK3CA/AKT/MAPK7/ERK in HT29 and HT29-5FU cells. The combination also inhibited EGFR, increased the preG1/S cell cycle phases, apoptosis, and caspase 8 in HT29 cells, while it increased the G1 phase, p21/p27, and apoptosis in HT29-5FU cells. Conclusion: We have combined the PamChip kinase profiling of Saudi CRC samples with in vitro drug combination studies in four CRC cells, highlighting the importance of targeting PIK3CA and ABCC1 for Saudi CRC patients, especially given that the overexpression of PIK3CA mutations was previously linked with the lack of activity for the anti-EGFRs as first line treatment for CRC patients. The combination of HAA2020 and 5FU has selectively sensitized the four CRC cells to 5FU and could be further studied.

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“…The vast majority of reports specified structure-based retrospective computational approaches, in which observed biological effects of compounds were underpinned mostly through molecular docking experiments with cryo-EM structures or homology models. Most pronounced are, again, ABCB1 [37] , [38] , [39] , [40] , [41] , [42] and ABCG2 [43] , [44] , [45] , [46] , [47] , [48] , [49] , [50] , [51] , [52] , [53] . Less- or under-studied ABC transporters are barely reflected in the literature.…”

Section: Introductionmentioning

confidence: 96%

Scaffold fragmentation and substructure hopping reveal potential, robustness, and limits of computer-aided pattern analysis (C@PA)

Namasivayam

Silbermann

Pahnke

et al. 2021

Computational and Structural Biotechnology Journal

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Sapitinib Reverses Anticancer Drug Resistance in Colon Cancer Cells Overexpressing the ABCB1 Transporter

Cited by 21 publications

References 59 publications

Overcoming multidrug resistance by knockout of ABCB1 gene using CRISPR/Cas9 system in SW620/Ad300 colorectal cancer cells

Overcoming multidrug resistance by knockout of ABCB1 gene using CRISPR/Cas9 system in SW620/Ad300 colorectal cancer cells

Chemosensitization of HT29 and HT29-5FU Cell Lines by a Combination of a Multi-Tyrosine Kinase Inhibitor and 5FU Downregulates ABCC1 and Inhibits PIK3CA in Light of Their Importance in Saudi Colorectal Cancer

Scaffold fragmentation and substructure hopping reveal potential, robustness, and limits of computer-aided pattern analysis (C@PA)

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