PMID- 30422397
OWN - NLM
STAT- MEDLINE
DCOM- 20200817
LR  - 20211204
IS  - 1552-4930 (Electronic)
IS  - 1552-4922 (Linking)
VI  - 95
IP  - 6
DP  - 2019 Jun
TI  - Concurrent detection of lysosome and tissue transglutaminase activation in 
      relation to cell cycle position during apoptosis induced by different anticancer 
      drugs.
PG  - 683-690
LID - 10.1002/cyto.a.23652 [doi]
AB  - Described is the new cytometric approach do detect either stimulation or a 
      collapse of lysosomal proton pump (lysosomes rupture) combined with activation of 
      transglutaminase 2 (TG2) during induction of apoptosis. Apoptosis of human 
      lymphoblastoid TK6 cells was induced by combination of 2-deoxyglucose with the 
      isoquinoline alkaloid berberine, by DNA topoisomerase I inhibitor camptothecin, 
      its analog topotecan, topoisomerase II inhibitors etoposide or mitoxantrone, as 
      well as by the cytotoxic anticancer ribonuclease ranpirnase (onconase). Activity 
      of the proton pump of lysosomes was assessed by measuring entrapment and 
      accumulation of the basic fluorochrome acridine orange (AO) resulting in its 
      metachromatic red luminescence (F(>640) ) within these organelles. Activation of 
      TG2 was detected in the same cell subpopulation by the evidence of crosslinking 
      of cytoplasmic proteins revealed by the increased intensity of the side light 
      scatter (SSC) as well as following cell lysis by detergent, by its red 
      fluorescence after staining by sulforhodamine 101. Because at low AO 
      concentration nuclear DNA of the lysed cells was stoichiometrically stained green 
      (F(530) ) its quantity provided information on effects of the drug treatments on 
      cell cycle in relation to activation of TG2. The data reveal that activation of 
      lysosomal proton pump was evident in subpopulations of cells treated with 
      2-deoxyglucose plus berberine, topotecan, etoposide and mitoxantrone but not with 
      ranpirnase. The collapse of lysosomal proton pump possibly reporting rupture of 
      these organelles was observed in definite cell subpopulations after treatment 
      with each of the studied drugs. Because regardless of the inducer of apoptosis 
      TG2 activation invariably was correlated with lysosomes rupture it is likely that 
      it was triggered by calcium ions or protons released from the ruptured lysosomes. 
      This new methodological approach offers the means to investigate mechanisms and 
      factors affecting autophagic lysosomes proton pump activity vis-a-vis TG2 
      activation that are common in several pathological states. (c) 2019 International 
      Society for Advancement of Cytometry.
CI  - (c) 2018 International Society for Advancement of Cytometry.
FAU - Halicka, H Dorota
AU  - Halicka HD
AD  - Department of Pathology, New York Medical College, Brander Cancer Research 
      Institute, Valhalla, New York.
FAU - Li, Jiangwei
AU  - Li J
AD  - Department of Pathology, New York Medical College, Brander Cancer Research 
      Institute, Valhalla, New York.
FAU - Zhao, Hong
AU  - Zhao H
AD  - Department of Pathology, New York Medical College, Brander Cancer Research 
      Institute, Valhalla, New York.
FAU - Darzynkiewicz, Zbigniew
AU  - Darzynkiewicz Z
AD  - Department of Pathology, New York Medical College, Brander Cancer Research 
      Institute, Valhalla, New York.
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20181113
PL  - United States
TA  - Cytometry A
JT  - Cytometry. Part A : the journal of the International Society for Analytical 
      Cytology
JID - 101235694
RN  - 0 (Antineoplastic Agents)
RN  - 0 (Proton Pumps)
RN  - EC 2.3.2.13 (Protein Glutamine gamma Glutamyltransferase 2)
RN  - EC 2.3.2.13 (Transglutaminases)
RN  - EC 3.6.1.- (GTP-Binding Proteins)
RN  - F30N4O6XVV (Acridine Orange)
SB  - IM
CIN - Cytometry A. 2019 Jul;95(7):712-713. PMID: 30382613
MH  - Acridine Orange/metabolism
MH  - Antineoplastic Agents/*pharmacology
MH  - Apoptosis/*drug effects
MH  - Autophagosomes/drug effects/enzymology
MH  - Cell Cycle/drug effects
MH  - Fluorescence
MH  - GTP-Binding Proteins/*metabolism
MH  - HL-60 Cells
MH  - Humans
MH  - Image Cytometry/*methods
MH  - Lysosomes/drug effects/*enzymology/metabolism
MH  - Protein Glutamine gamma Glutamyltransferase 2
MH  - Proton Pumps/*drug effects/metabolism
MH  - Transglutaminases/*metabolism
OTO - NOTNLM
OT  - acridine orange
OT  - autophagy
OT  - lysosomes rupture
OT  - protein crosslinking
OT  - proton pump
OT  - sulforhodamine 101
EDAT- 2018/11/14 06:00
MHDA- 2020/08/18 06:00
CRDT- 2018/11/14 06:00
PHST- 2018/07/30 00:00 [received]
PHST- 2018/09/18 00:00 [revised]
PHST- 2018/09/28 00:00 [accepted]
PHST- 2018/11/14 06:00 [pubmed]
PHST- 2020/08/18 06:00 [medline]
PHST- 2018/11/14 06:00 [entrez]
AID - 10.1002/cyto.a.23652 [doi]
PST - ppublish
SO  - Cytometry A. 2019 Jun;95(6):683-690. doi: 10.1002/cyto.a.23652. Epub 2018 Nov 13.