VSports app下载 - Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The . gov means it’s official. Federal government websites often end in . gov or VSports app下载. mil. Before sharing sensitive information, make sure you’re on a federal government site. .

Https

The site is secure V体育官网. The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely. .

. 2016 Mar;30(3):761-4.
doi: 10.1038/leu.2015.184. Epub 2015 Jul 15.

"V体育安卓版" BH3 profiling identifies heterogeneous dependency on Bcl-2 family members in multiple myeloma and predicts sensitivity to BH3 mimetics

C Touzeau  1   2   3 , J Ryan  1 , J Guerriero  1 , P Moreau  2   3 , T N Chonghaile  4 , S Le Gouill  2   3 , P Richardson  1 , K Anderson  1 , M Amiot  3 , A Letai  1
Affiliations

BH3 profiling identifies heterogeneous dependency on Bcl-2 family members in multiple myeloma and predicts sensitivity to BH3 mimetics

C Touzeau et al. Leukemia. 2016 Mar.
No abstract available

PubMed Disclaimer

Conflict of interest statement

CONFLICT-OF-INTEREST DISCLOSURE

AL has been a paid advisor to AbbVie. AL’s laboratory has received sponsorship for research with AbbVie VSports手机版.

V体育ios版 - Figures

Figure 1
Figure 1. BH3 profiling of Myeloma cell lines demonstrates heterogeneous Bcl-2 dependancy and correlates with in vitro sensitivity to BH3 mimetics
(a) Binding affinity of BH3 peptides, ABT-199 and ABT-263 for the anti-apoptotic proteins. Green and red colors indicate high and low affinity, respectively,,. (b) KMS12-PE, XG-5, SKMM2, LP-1, L363, NCI-H929, RPMI-8226 and MM1-S identity was confirmed by DNA fingerprinting or HLA typing. Intracellular BH3 (iBH3) profiling was performed using BAD (100µM), HRK (100µM) and MS-1 (10µM) peptides. As previously described , tumor cells were pelleted and resuspended in DTEB buffer with addition of each BH3 peptide treatment with 0.002% w/v digitonin. BAD, HRK and MS1 peptides sequences have been previously described,. Mitochondria in the permeabilized cells were exposed to peptides for 45 min at 26°C before fixation with 2% formaldehyde at room temperature for 15 min. After addition of neutralizing buffer, cells were stained with anti-cytochrome c–Alexa488 (#560263, BD Biosciences) 1:100 in 0.1% Saponin/1%BSA/PBS overnight at 4 C. The quantification of cytochrome c loss induced by each peptide was analyzed by FACS (LSR Fortessa flow cytometer, BD Bioscience). Values indicate the percentage of cytochrome c negative cells. Indicated values are the mean of 3 independent experiments. (c) Sensitivity of Myeloma cell lines to the BH3 mimetics ABT-199 and ABT-263 was assessed using Cell Titer Glow® assay. Cell lines were cultured with increasing doses of ABT-199 or ABT-263 during 24 hours. Indicated values are the mean of 3 independent experiments. Light grey line indicates IC50 <100 nM, Dark grey line indicates IC50 between 100 and 5000 nM, black lines indicate IC50 > 5000 nM (d) IC50 of myeloma cell lines were correlated with the mitochondrial response to BAD (100µM), BAD (100µM) – HRK (100µM) peptides and with ABT-199 or 263 (1µM). All correlations were tested using a one-tailed Spearman r correlation using GraphPad Prism software.
Figure 2
Figure 2. BH3 profiling of primary myeloma cells predicts lines in vitro sensitivity to BH3 mimetics
(a) Mitochondrial priming of multiple myeloma primary cells. Primary cells were obtained after informed consent from MM patients treated at the Dana Farber Cancer Institute, Boston, USA. Intracellular BH3 (iBH3) profiling was performed using BAD (100µM), HRK (100µM) and MS-1 (10µM) peptides. Abreviations:Iso, immunoglobulin isotype; Diag, Newly diagnosed myeloma; Rel, relapsed myeloma. HD: hyperdiploidy. 1All samples were screened by FISH for the t(11;14), t(4;14), 17p deletion and caryotype was performed for hyperdiploidy status. 2Values indicate the percentage of cytochrome c negative cell. 3Values indicate the percentage of apoptotic cells. Cells were cultured with/without the drugs during 16 hours. Cell death was assessed by flow cytometry after annexin V staining Dark grey: ≥50%, Intermediate grey: 20–49, light grey: <20% (b) Primary samples were cultured for 16 hours with/without ABT-199 or ABT-263 (100 nM). Cell death was measured by FACS using Annexin V staining. Percentages of apoptotic cells were correlated with the BAD (100µM) peptide. All correlations were tested using a one-tailed Spearman r correlation using GraphPad Prism software.
See this image and copyright information in PMC

References

    1. Letai AG. Diagnosing and exploiting cancer’s addiction to blocks in apoptosis. Nat Rev Cancer. 2008;8:121–132. - PubMed
    1. Certo M, Del Gaizo Moore V, Nishino M, Wei G, Korsmeyer S, Armstrong SA, et al. Mitochondria primed by death signals determine cellular addiction to antiapoptotic BCL-2 family members. Cancer Cell. 2006;9:351–365. - PubMed
    1. BC-2 inhibitor yields high response in CLL and SLL. Cancer Discov. 2014;4:OF5. - PubMed
    1. Davids MS, Roberts AW, Anderson MA, Pagel JM, Kahl BS, Gerecitano JF, et al. The BCL-2-Specific BH3-Mimetic ABT-199 (GDC-0199) Is Active and Well-Tolerated in Patients with Relapsed Non-Hodgkin Lymphoma: Interim Results of a Phase I Study. ASH Annu Meet Abstr. 2012;120:304.
    1. Souers AJ, Leverson JD, Boghaert ER, Ackler SL, Catron ND, Chen J, et al. ABT-199, a potent and selective BCL-2 inhibitor, achieves antitumor activity while sparing platelets. Nat Med. 2013 - PubMed

MeSH terms