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

Lei, Zi‐Ning; Teng, Quincy; Wu, Zhuo‐Xun; Ping, Fengfeng; Song, Peng; Wurpel, John N.D.; Chen, Zhe‐Sheng

doi:10.1002/mco2.106

Cited by 30 publications

(19 citation statements)

References 39 publications

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“…Treatment with P-gp modulators can provide more significant cytotoxicity by reducing effective concentrations by 100-fold (paclitaxel) or 351-fold (vinblastine) in MDR colorectal cancer SW620/Ad300 cells [31,32] .…”

Section: Discussionmentioning

confidence: 99%

Multimodal 4-arylchromene derivatives with microtubule-destabilizing, anti-angiogenic, and MYB-inhibitory activities

Köhler¹,

Reich²,

Yusenko³

et al. 2023

Cancer Drug Resist

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Aim: Efficient and readily available anticancer drugs are sought as treatment options. For this reason, chromene derivatives were prepared using the one-pot reaction and tested for their anticancer and anti-angiogenic properties. Methods: 2-Amino-3-cyano-4-(aryl)-7-methoxy-4H-chromene compounds (2A-R) were repurposed or newly synthesized via a three-component reaction of 3-methoxyphenol, various aryl aldehydes, and malononitrile. We performed assays to study the inhibition of tumor cell growth (MTT assay), effects on microtubules (immunofluorescence), cell cycle (flow-activated cell sorting analysis), angiogenesis (zebrafish model), and MYB activity (luciferase reporter assay). Fluorescence microscopy was applied for localization studies via copper-catalyzed azide-alkyne click reaction of an alkyne-tagged drug derivative. Results: Compounds 2A-C and 2F exhibited robust antiproliferative activities against several human cancer cell lines (50% inhibitory concentrations in the low nanomolar range) and showed potent MYB inhibition. The alkyne derivative 3 was localized in the cytoplasm after only 10 min of incubation. Substantial microtubule disruption and G2/M cell-cycle arrest were observed, where compound 2F stood out as a promising microtubule-disrupting agent. The study of anti-angiogenic properties showed that 2A was the only candidate with a high potential to inhibit blood vessel formation in vivo. Conclusion: The close interplay of various mechanisms, including cell-cycle arrest, MYB inhibition, and anti-angiogenic activity, led to identifying promising multimodal anticancer drug candidates.

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Section: Discussionmentioning

confidence: 99%

Multimodal 4-arylchromene derivatives with microtubule-destabilizing, anti-angiogenic, and MYB-inhibitory activities

Köhler¹,

Reich²,

Yusenko³

et al. 2023

Cancer Drug Resist

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“…Second, although artificially engineered nucleases have greatly improved the efficiency of genome editing, homology-directed repair (HDR)-mediated mutation replacement remains difficult in human cells, especially iPSCs without selection markers [ 22 , 23 , 24 ]. The efficiency of nonhomologous end joining (NHEJ) is much higher than that of HDR [ 25 , 26 , 27 ]. Therefore, Jin-jing et al described Cas9-mediated splicing regulatory element disruption with 54 analyzed clones and 13 clones with NHEJ.…”

Section: Discussionmentioning

confidence: 99%

Targeted-Deletion of a Tiny Sequence via Prime Editing to Restore SMN Expression

Zhou

Tang

Duan

et al. 2022

IJMS

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Spinal muscular atrophy (SMA) is a devastating autosomal recessive motor neuron disease associated with mutations in the survival motor neuron 1 (SMN1) gene, the leading genetic cause of infant mortality. A nearly identical copy gene (SMN2) is retained in almost all patients with SMA. However, SMN2 fails to prevent disease development because of its alternative splicing, leading to a lack of exon 7 in the majority of SMN2 transcripts and yielding an unstable truncated protein. Several splicing regulatory elements, including intronic splicing silencer-N1 (ISS-N1) of SMN2 have been described. In this study, targeted-deletion of ISS-N1 was achieved using prime editing (PE) in SMA patient-specific induced pluripotent stem cells (SMA-iPSCs) with a high efficiency of 7/24. FL-SMN expression was restored in the targeted-deletion iPS clones and their derived motor neurons (iMNs). Notably, the apoptosis of the iMNs, caused by the loss of SMN protein that leads to the hyperactivity of endoplasmic reticulum (ER) stress, was alleviated in targeted-deletion iPSCs derived-iMNs. Thus, this is the first study to demonstrate that the targeted-deletion of ISS-N1 via PE for restoring FL-SMN expression holds therapeutic promise for SMA.

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“…The human colon cancer cell line, SW620, was used as the parental cell line, and the doxorubicin-selected subline, SW620/Ad300, which overexpresses the ABCB1 transporter ( Lai et al, 1991a ; Lai et al, 1991b ), was used as another drug-resistant cell line. We also used SW620 parental and SW620/Ad300 drug-resistant cell sublines, where the gene for the ABCB1 transporter was knocked out using the CRISPR/Cas9 system ( Lei et al, 2021 ). The human large cell lung cancer cell line, NCI-H460, was used as the parental cell line and its mitoxantrone-selected subline, NCI-H460/MX20, which overexpresses the ABCG2 transporter ( Henrich et al, 2007 ), was used as another drug-resistant cell line.…”

Section: Methodsmentioning

confidence: 99%

The Resistance of Cancer Cells to Palbociclib, a Cyclin-Dependent Kinase 4/6 Inhibitor, is Mediated by the ABCB1 Transporter

Lei

et al. 2022

Front. Pharmacol.

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Palbociclib was approved by the United States Food and Drug Administration for use, in combination with letrozole, as a first-line treatment for estrogen receptor-positive/human epidermal growth factor receptor 2-negative (ER+/HER2-) postmenopausal metastatic breast cancer. However, recent studies show that palbociclib may be an inhibitor of the ABCB1 transporter, although this remains to be elucidated. Therefore, we conducted experiments to determine the interaction of palbociclib with the ABCB1 transporter. Our in vitro results indicated that the efficacy of palbociclib was significantly decreased in the ABCB1-overexpressing cell lines. Furthermore, the resistance of ABCB1-overexpressing cells to palbociclib was reversed by 3 μM of the ABCB1 inhibitor, verapamil. Moreover, the incubation of ABCB1-overexpressing KB-C2 and SW620/Ad300 cells with up to 5 μM of palbociclib for 72 h, significantly upregulated the protein expression of ABCB1. The incubation with 3 µM of palbociclib for 2h significantly increased the intracellular accumulation of [3H]-paclitaxel, a substrate of ABCB1, in ABCB1 overexpressing KB-C2 cells but not in the corresponding non-resistant parental KB-3-1 cell line. However, the incubation of KB-C2 cells with 3 μM of palbociclib for 72 h decreased the intracellular accumulation of [3H]-paclitaxel due to an increase in the expression of the ABCB1 protein. Palbociclib produced a concentration-dependent increase in the basal ATPase activity of the ABCB1 transporter (EC50 = 4.73 μM). Molecular docking data indicated that palbociclib had a high binding affinity for the ABCB1 transporter at the substrate binding site, suggesting that palbociclib may compete with other ABCB1 substrates for the substrate binding site of the ABCB1. Overall, our results indicate that palbociclib is a substrate for the ABCB1 transporter and that its in vitro anticancer efficacy is significantly decreased in cancer cells overexpressing the ABCB1.

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

Cited by 30 publications

References 39 publications

Multimodal 4-arylchromene derivatives with microtubule-destabilizing, anti-angiogenic, and MYB-inhibitory activities

Multimodal 4-arylchromene derivatives with microtubule-destabilizing, anti-angiogenic, and MYB-inhibitory activities

Targeted-Deletion of a Tiny Sequence via Prime Editing to Restore SMN Expression

The Resistance of Cancer Cells to Palbociclib, a Cyclin-Dependent Kinase 4/6 Inhibitor, is Mediated by the ABCB1 Transporter

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