PMID- 30397244
OWN - NLM
STAT- MEDLINE
DCOM- 20191022
LR  - 20191022
IS  - 2045-2322 (Electronic)
IS  - 2045-2322 (Linking)
VI  - 8
IP  - 1
DP  - 2018 Nov 5
TI  - Comparison of cell-based assays to quantify treatment effects of anticancer drugs 
      identifies a new application for Bodipy-L-cystine to measure apoptosis.
PG  - 16363
LID - 10.1038/s41598-018-34696-x [doi]
LID - 16363
AB  - Cell-based assays that measure anticancer drug effects are essential for 
      evaluating chemotherapeutic agents. Many assays targeting various cellular 
      mechanisms are available, leading to inconsistent results when using different 
      techniques. We critically compared six common assays, as well as a new assay 
      using Bodipy.FL.L-cystine (BFC), to identify the most accurate and reproducible 
      in measuring anticancer drug effects. We tested three common chemotherapies 
      (methotrexate, paclitaxel, and etoposide) in two cell lines (Ln229 and 
      MDA-MB231). Spectroscopic assays such as Cell Titer Blue, and 
      2',7'-dichlorofluorescin diacetate (DCFDA) yielded a strong drug dose response, 
      especially for paclitaxel and etoposide (R(2) = 0.9). MTT and Calcein-AM 
      fluorescent dye-based assays were less consistent in that regard. Among three 
      flow cytometry assays, Propidium Iodide (PI)-based DNA content analysis and a new 
      BFC-based glutathione-redox (GSH) assay produced drug dose dependent results. 
      Compared to PI, BFC showed a better correlation (R(2) = 0.7-0.9) in depicting 
      live and apoptotic cells. We found that the combination of Cell Titer Blue 
      spectroscopy and BFC flow cytometry assays were most accurate in assessing 
      anticancer drug effects by clear distinction between live and apoptotic cells, 
      independent of drug mechanism of action. We present a new application of BFC as 
      an agent for measuring cellular apoptosis.
FAU - Kumar, Nita
AU  - Kumar N
AD  - Cellular Pathway Imaging Laboratory (CPIL), Molecular Imaging Program at 
      Stanford, Stanford University School of Medicine, 3155 Porter Drive, Palo Alto, 
      CA, 94305, USA.
FAU - Afjei, Rayhaneh
AU  - Afjei R
AD  - Laboratory of Experimental and Molecular Neuroimaging (LEMNI), Molecular Imaging 
      Program at Stanford, Stanford University School of Medicine, 300 Pasteur Drive, 
      Grant S-031, Stanford, CA, 94305, USA.
FAU - Massoud, Tarik F
AU  - Massoud TF
AD  - Laboratory of Experimental and Molecular Neuroimaging (LEMNI), Molecular Imaging 
      Program at Stanford, Stanford University School of Medicine, 300 Pasteur Drive, 
      Grant S-031, Stanford, CA, 94305, USA.
FAU - Paulmurugan, Ramasamy
AU  - Paulmurugan R
AD  - Cellular Pathway Imaging Laboratory (CPIL), Molecular Imaging Program at 
      Stanford, Stanford University School of Medicine, 3155 Porter Drive, Palo Alto, 
      CA, 94305, USA. paulmur8@stanford.edu.
LA  - eng
GR  - R01 CA209888/CA/NCI NIH HHS/United States
GR  - R21 EB022298/EB/NIBIB NIH HHS/United States
GR  - R25 CA217729/CA/NCI NIH HHS/United States
PT  - Comparative Study
PT  - Journal Article
PT  - Research Support, N.I.H., Extramural
DEP - 20181105
PL  - England
TA  - Sci Rep
JT  - Scientific reports
JID - 101563288
RN  - 0 (Amino Acid Transport Systems, Acidic)
RN  - 0 (Antineoplastic Agents)
RN  - 0 (BODIPY-FL L-cystine)
RN  - 0 (Boron Compounds)
RN  - 0 (Tumor Suppressor Protein p53)
RN  - 48TCX9A1VT (Cystine)
SB  - IM
MH  - Amino Acid Transport Systems, Acidic/metabolism
MH  - Antineoplastic Agents/*pharmacology
MH  - Apoptosis/*drug effects
MH  - Boron Compounds/*pharmacology
MH  - Cell Line, Tumor
MH  - Cell Survival/drug effects
MH  - Cystine/*analogs & derivatives/pharmacology
MH  - Dose-Response Relationship, Drug
MH  - Drug Screening Assays, Antitumor/*methods
MH  - Humans
MH  - Time Factors
MH  - Tumor Suppressor Protein p53/metabolism
PMC - PMC6218539
COIS- The authors declare no competing interests.
EDAT- 2018/11/07 06:00
MHDA- 2019/10/23 06:00
PMCR- 2018/11/05
CRDT- 2018/11/07 06:00
PHST- 2018/05/01 00:00 [received]
PHST- 2018/10/18 00:00 [accepted]
PHST- 2018/11/07 06:00 [entrez]
PHST- 2018/11/07 06:00 [pubmed]
PHST- 2019/10/23 06:00 [medline]
PHST- 2018/11/05 00:00 [pmc-release]
AID - 10.1038/s41598-018-34696-x [pii]
AID - 34696 [pii]
AID - 10.1038/s41598-018-34696-x [doi]
PST - epublish
SO  - Sci Rep. 2018 Nov 5;8(1):16363. doi: 10.1038/s41598-018-34696-x.