Comparative analysis of apoptosis in HL60 detected by annexin-V and fluorescein-diacetate

Bartkowiak, Detlef; Högner, Stephanie; Baust, Heinrich; Nothdurft, Wilhelm; Röttinger, E. M.

doi:10.1002/(sici)1097-0320(19991101)37:3<191::aid-cyto5>3.0.co;2-u

Cited by 49 publications

(22 citation statements)

References 30 publications

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“…Apoptotic cell enumeration was evaluated by the amount of cells with the cell membrane impermeable for PI and decreased amount of fluorescein [23]. Approximately 5 x 10 5 untreated or treated cells were resuspended in 400 μl of PBS/0.2% BSA containing 10 nM of FDA at room temperature, for 30 min.…”

Section: Methodsmentioning

confidence: 99%

Proteasome inhibition leads to altered signaling in the proteome of cisplatin-resistant human ovarian carcinoma cell line

Duraj¹,

Pastorek²,

Vitkovska³

et al. 2014

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To address a precise view into molecular mechanisms of apoptotic signaling pathways after single-or combinatory treatments with specific NF-κB-or proteasome inhibitors and/or cisplatin (CDDP), flow cytometry and western blotting of the cell proteome in human ovarian chemosensitive-and CDDP-resistant cell lines were used. We report here that proteasome inhibition (but not NF-κB inhibition) caused marked alterations in the cell proliferation and cell cycle, as well as in the levels of signaling anti-and pro-apoptotic proteins PARP, NF-κB, IκB-α, Bcl-2, Bax, and lysosome-associated LAMP-1 and ATP-7B molecules in particular proteome fractions. These findings refer to the possibility of regulation of CDDP resistance, inclusive the capacity of lysosomes to export CDDP in certain human ovarian cancer cells by proteasome inhibition.

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Section: Methodsmentioning

confidence: 99%

Proteasome inhibition leads to altered signaling in the proteome of cisplatin-resistant human ovarian carcinoma cell line

Duraj¹,

Pastorek²,

Vitkovska³

et al. 2014

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“…Apoptotic cell enumeration was done according to the amount of cells with the cell membrane impermeable for PI with low amount of fluorescein, or cells with sub-G 1 amount of DNA after detergent permeabilization. As for FDA/PI staining [26], both cell lines were collected as described above. Pooled cells were washed twice with cold PBS.…”

Section: Methodsmentioning

confidence: 99%

Synthetic isothiocyanate indole-3-ethyl isothiocyanate (homoITC) enhances sensitivity of human ovarian carcinoma cell lines A2780 and A2780/CP to cisplatin

Stehlik

Paulíková²,

Hunáková³

2010

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Isothiocyanates (ITCs), popular chemopreventive agents present in cruciferous vegetables, prove growth-inhibiting and apoptosis-inducing activities in cancer cell lines in vitro. Our study presents a new synthetic ITC derivate indol-3-ethyl isothiocyanate (homoITC) as an effective modulator of cellular proliferation and inducer of apoptosis with potential utility as an anticancer drug or a sensitizer to routinely used chemotherapeutic agent cisplatin (cis-Pt).We analyzed the growth inhibitory effects of homoITC in the human ovarian carcinoma cell line A2780 and its cisplatinresistant variant A2780/CP using MTT-test and its apoptosis-inducing properties by flow cytometry and caspase 3 activation. Combination index (CI) values from Calcusyn software were used to characterize the interactions of homoITC and cis-Pt as synergistic (CI<1), additive (CI=1), or antagonistic (CI>1). Significant synergistic effect in growth inhibition of homoITC (5 -15 µM) and cis-Pt (2.5 -10 µM) on A2780 parental cell line (CI from 0.42 to 0.85) was also observed on A2780/CP resistant subline (CI from 0.18 to 0.73) for 10-50 µM cis-Pt concentrations and the same concentrations of homoITC. Synergy in growth inhibition correlated with the potential of homoITC to stimulate apoptosis induced by cis-Pt.We conclude that homoITC may be worth of further studies assessing its value in the ovarian cancer treatment and elucidating mechanisms of its action.

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“…On the test day, adherent cells were harvested by trypsinization and recombined with the culture supernatant, containing detached, i.e., presumably dead cells. The samples were stained with PI (5 µg/ml) and fluorescein diacetate (FDA, 10 nM) and analyzed by flow cytometry as described previously [3].…”

Section: Genementioning

confidence: 99%

Radiation- and Chemoinduced Multidrug Resistance in Colon Carcinoma Cells

et al. 2009

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Comparative analysis of apoptosis in HL60 detected by annexin-V and fluorescein-diacetate

Cited by 49 publications

References 30 publications

Proteasome inhibition leads to altered signaling in the proteome of cisplatin-resistant human ovarian carcinoma cell line

Proteasome inhibition leads to altered signaling in the proteome of cisplatin-resistant human ovarian carcinoma cell line

Synthetic isothiocyanate indole-3-ethyl isothiocyanate (homoITC) enhances sensitivity of human ovarian carcinoma cell lines A2780 and A2780/CP to cisplatin

Radiation- and Chemoinduced Multidrug Resistance in Colon Carcinoma Cells

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