Anti-cell growth and anti-cancer stem cell activity of the CDK4/6 inhibitor palbociclib in breast cancer cells

Kishino, Emi; Ogata, Ryohei; Saitoh, Wataru; Koike, Yoshikazu; Ohta, Yusuke; Kanomata, Naoki; Kurebayashi, Junichi

doi:10.1007/s12282-019-01035-5

Cited by 14 publications

(21 citation statements)

References 19 publications

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“…For the E2-sensitivity assay, the cells were cultured in the estrogen-deprived medium plus the indicated concentrations of E2 for 3 days. Then, the cells were harvested and counted using a Coulter counter (Coulter Electronics, Harpenden, UK) [7].…”

Section: Cell Growth Assaymentioning

confidence: 99%

“…To investigate cell cycle progression, harvested cells were stained with propidium iodide using the CycleTest Plus DNA Reagent kit (Becton-Dickinson, San Jose, CA, USA) according to the manufacturer's recommendations. Flow cytometry was performed using a FACSCalibur flow cytometer (Becton-Dickinson) and the DNA histogram was analyzed using CELLQuest version 6.0 (Becton-Dickinson) [7].…”

Section: Cell Cycle and Cell Senescence Assaysmentioning

confidence: 99%

“…Cells were then washed with PBS, fixed and stained with the SA-β gal solution (Millipore) for 4 hours or overnight. Senescent cells were quantified by counting 100 cells in 3 different fields for each replicate [7].…”

Section: Cell Cycle and Cell Senescence Assaysmentioning

confidence: 99%

“…Labeled protein was visualized using the ECL Prime Western Blotting Detection Reagent (GE Healthcare Japan, Tokyo, Japan), with the expression of β-actin as the internal standard. The expression levels were measured using Quantity One 1-D software ver.4.5 (BIO-RAD, Tokyo, Japan) [7].…”

Section: Western Blot Analysismentioning

confidence: 99%

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Resistance to cyclin-dependent kinase (CDK) 4/6 inhibitors confers cross-resistance to other CDK inhibitors but not to chemotherapeutic agents in breast cancer cells

et al. 2020

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Background Combined endocrine therapy with a cyclin-dependent kinase (CDK) 4/6 inhibitor has been indicated to improve not only progression-free survival, but also overall survival in patients with hormone receptor (HR)-positive, HER2-negative advanced breast cancer. However, resistance to this combination therapy inevitably develops. How to manage this resistant breast cancer is one of the most important clinical issues. To investigate the mechanisms of action responsible for resistance, we developed breast cancer cells resistant to CDK4/6 inhibitors, and analyzed their biological characteristics and sensitivity to different anticancer agents. Methods HR-positive, HER2-negative MCF-7 and KPL-1 breast cancer cells were cultivated in palbociclib (PAL) or abemaciclib (ABE)-added culture medium for over 5 months, and we successfully developed PAL-or ABE-resistant cells. The effects of PAL or ABE on the cell growth, basal RB expression, RB phosphorylation, cell cycle and cell senescence were compared between resistant and parental cells. Effects of the other CDK4/6 inhibitor, different chemotherapeutic agents and estrogen on the cell growth were also examined. The expression levels of cyclin D1, CDK2, CDK4, CDK6, cyclin E1 and estrogen receptor (ER)-ɑ were measured using RT-PCR. Results Long-term exposure to up to 200 nM PAL or ABE resulted in the development of PAL-or ABE-resistant MCF-7 or KPL-1 breast cancer cells. Basal expression levels of RB in both resistant cells were down-regulated. Inhibitory effects of either PAL or ABE on RB phosphorylation were reduced in both resistant cells. Accordingly, G1-S cell cycle retardation and cell senescence induced by either inhibitor were also attenuated in both resistant cells. Both resistant cells were crossresistant to the other CDK4/6 inhibitor but almost as equally sensitive to different chemotherapeutic agents (5-fluorouracil, gemcitabine, paclitaxel, docetaxel, doxorubicin and eribulin) as the parental cells. The mRNA expression level of CDK6 significantly increased in the resistant MCF-7 cells and that of Rb1 significantly decreased in the resistant KPL-1 cells. Although both resistant cells were less sensitive to estrogen than the parental cells, the expression levels of ER-ɑ did not significantly change in either. Conclusions Our study suggests that acquired resistance to PAL or ABE confers cross-resistance to the other CDK4/6 inhibitor but not to chemotherapeutic agents in HR-positive, HER2-negative breast cancer cells. Down-regulation of basal RB expression and normalized RB phosphorylation reduced by CDK4/6 inhibitors may be responsible for the attenuated anti-cell growth effects of the inhibitors.

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Section: Cell Growth Assaymentioning

confidence: 99%

Section: Cell Cycle and Cell Senescence Assaysmentioning

confidence: 99%

Section: Cell Cycle and Cell Senescence Assaysmentioning

confidence: 99%

Section: Western Blot Analysismentioning

confidence: 99%

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Resistance to cyclin-dependent kinase (CDK) 4/6 inhibitors confers cross-resistance to other CDK inhibitors but not to chemotherapeutic agents in breast cancer cells

et al. 2020

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“…Several studies have shown that gastrointestinal reactions, including abdominal pain, diarrhea, bloating, are the most common adverse reactions to tyrosine kinase inhibitors [13][14][15][16]. Therefore, tyrosine kinase inhibitors are often recommended to be administrated with food.…”

mentioning

confidence: 99%

Effect of high-fat diet on the pharmacokinetics and safety of flumatinib in healthy Chinese subjects

Kuang

Song

Yang

et al. 2020

Cancer Chemother Pharmacol

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Purpose To evaluate the effect of a high-fat diet on the pharmacokinetics and safety of flumatinib mesylate tablets in healthy Chinese subjects. Methods This study was a randomized, open-label, single-dose, two-period crossover trial in which subjects were randomly assigned to take 400 mg of flumatinib mesylate after a high-fat diet or a fasted state. After a 14-day washout period, the two groups were administered flumatinib mesylate under opposite conditions. Blood samples were collected at baseline 0 and 0.5, 1, 1.5, 2, 2.5, 3, 4, 5, 6, 8, 10, 12, 24, 48, 72, and 96 h, respectively. Plasma concentrations of flumatinib and its metabolites (M1 and M3) were analyzed using liquid chromatography-mass spectrometry. Pharmacokinetic parameters were calculated using the non-compartmental module of the Phoenix WinNonlin Version 7.0 software. BE module of WinNonLin was used for statistical analysis of AUC0–t, AUC0–∞ and Cmax in plasma. Results Twelve healthy subjects, half male and half female, were enrolled. One subject withdrew due to a treatment-emergent adverse event. Eleven subjects were administered drugs on fasting and 12 were administered drugs after a high-fat diet. On high-fat diet/fasting, the least square geometric mean (LSGM) ratios of flumatinib, M1, M3, and their 90% confidence interval (CI) were as follows: for flumatinib, Cmax, AUC0–t and AUC0–∞ were 281.65% (225.80–351.31%), 167.43% (143.92–194.79%), and 166.87% (143.47–194.09%); for M1, Cmax, AUC0–t, and AUC0–∞ were 188.59% (145.29–244.79), 163.94% (149.11–180.24%), and 164.48% (150.36–179.94%); for M3, Cmax, AUC0–t, and AUC0–∞ were 63.47% (54.02–74.57%), 85.23% (74.72–97.22%), and 96.73% (86.63–108.02%). Conclusion Among the subjects, oral administration of 400 mg of flumatinib was safe and well tolerated. High-fat diet significantly increases the exposure to flumatinib, therefore, fasting may be recommended. Clinical trial registration The study was registered at chictr.org Identifier: ChiCTR-IIR-17013179.

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Role of cyclins and cyclin-dependent kinases in pluripotent stem cells and their potential as a therapeutic target

Jirawatnotai

Dalton

Wattanapanitch

2020

Seminars in Cell & Developmental Biology

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Anti-cell growth and anti-cancer stem cell activity of the CDK4/6 inhibitor palbociclib in breast cancer cells

Cited by 14 publications

References 19 publications

Resistance to cyclin-dependent kinase (CDK) 4/6 inhibitors confers cross-resistance to other CDK inhibitors but not to chemotherapeutic agents in breast cancer cells

Resistance to cyclin-dependent kinase (CDK) 4/6 inhibitors confers cross-resistance to other CDK inhibitors but not to chemotherapeutic agents in breast cancer cells

Effect of high-fat diet on the pharmacokinetics and safety of flumatinib in healthy Chinese subjects

Role of cyclins and cyclin-dependent kinases in pluripotent stem cells and their potential as a therapeutic target

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