CoChat
Sign Up
Cited by 76

The DOI matches its Crossref record; 2 of the 2 references it cites were found in a registry or an index.

Autophagy-deficient breast cancer shows early tumor recurrence and escape from dormancy

Aqbi, Hussein F., Tyutyunyk-Massey, Liliya, Keim, Rebecca C., Butler, Savannah E., Thekkudan, Theresa, Joshi, Supriya, Smith, Timothy M., Bandyopadhyay, Dipankar, Idowu, Michael O., Bear, Harry D., Payne, Kyle K., Gewirtz, David A., Manjili, Masoud H.·2018·Oncotarget
DOI 10.18632/oncotarget.25197PMC PMC5955162
Open this paper in CoChat — free account

Overview

The claims and statistics of this paper haven’t been checked yet.

Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.

A few free checks a day · papers already verified open instantly · results are public

Integrity signals

Registry

Crossref · 10.18632/oncotarget.25197

Cites

2 of 2 references verified0 citation errors · 0 not checkable

Statistics

Figures

Figure 1 CQ blocks ADR-induced autophagy MMC tumor cells received three daily doses of ADR alone (1 µM ADR for 2 hrs) (ADR) or in the presence of CQ (10 µM 3 hrs before ADR and 2 hrs during ADR treatment) (ADR+CQ), washed after each daily treatment and analyzed by acridine orange (AO) one day after the last treatment. Untreated MMC (Medium) or MMC treated with CQ (CQ) served as controls. (A) Acridine orange (AO) staining was analyzed for acidic vesicles (red) using image cytometry. Data represent triplicate experiments. (B) Levels of p62/SQSTM1 and LC.3B after treatment with ADR ± CQ indicative of autophagy induction in the absence of autophagic flux (B).
Figure 1 CQ blocks ADR-induced autophagy MMC tumor cells received three daily doses of ADR alone (1 µM ADR for 2 hrs) (ADR) or in the presence of CQ (10 µM 3 hrs before ADR and 2 hrs during ADR treatment) (ADR+CQ), washed after each daily treatment and analyzed by acridine orange (AO) one day after the last treatment. Untreated MMC (Medium) or MMC treated with CQ (CQ) served as controls. (A) Acridine orange (AO) staining was analyzed for acidic vesicles (red) using image cytometry. Data represent triplicate experiments. (B) Levels of p62/SQSTM1 and LC.3B after treatment with ADR ± CQ indicative of autophagy induction in the absence of autophagic flux (B).CC BY · PubMed Central
Figure 2 ADR-induced dormant tumor cells remain dormant in the presence of CQ MMC tumor cells were treated with 3 daily doses of ADR (1 uM for 2 hrs), with one group receiving CQ (10 uM) 3 hrs prior to and during ADR treatment. Both groups remained untreated for 3 weeks and 6 weeks, in vitro. (A) Adherent viable cells were counted using trypan blue exclusion at various time points. Data represent 3 replicates ± SEM. (B) At weeks 3 and 6 post-treatment, Ki-67 expression (upper panel) and viability (lower panel) were quantified within the population of adherent tumor cells. Data represent 2–3 replicates ± SEM. Four independent experiments have been carried out which have shown similar results.
Figure 2 ADR-induced dormant tumor cells remain dormant in the presence of CQ MMC tumor cells were treated with 3 daily doses of ADR (1 uM for 2 hrs), with one group receiving CQ (10 uM) 3 hrs prior to and during ADR treatment. Both groups remained untreated for 3 weeks and 6 weeks, in vitro. (A) Adherent viable cells were counted using trypan blue exclusion at various time points. Data represent 3 replicates ± SEM. (B) At weeks 3 and 6 post-treatment, Ki-67 expression (upper panel) and viability (lower panel) were quantified within the population of adherent tumor cells. Data represent 2–3 replicates ± SEM. Four independent experiments have been carried out which have shown similar results.CC BY · PubMed Central
Figure 3 Dormant tumor cells established by ADR or ADR+CQ remain susceptible to immunotherapy The in vitro tumor dormancy was established three weeks after three daily treatments of MMC with ADR or ADR+CQ. Untreated MMC cells were used as control. (A) Apoptosis was determined by FVS viability staining in MMC (control), ADR-treated dormant MMC (ADR), ADR+CQ-treated dormant MMC (ADR+CQ), as well as control MMC cultured with three daily doses of IFN-g and analyzed two days later (50 ng/ml) (IFN-g), ADR-treated dormant MMC cultured with three daily doses of IFN-g (50 ng/ml) and analyzed two days later (ADR > IFN-g), or ADR+CQ-treated dormant MMC cultured with three daily doses of IFN-g (50 ng/ml) and analyzed two days later (ADR+CQ > IFN-g). (B) Apoptosis was determined by FVS viability staining of MMC (control), MMC cultured with MMC-sensitized T cells for 48 hrs (T cells), ADR-treated dormant MMC (ADR), ADR-treated dormant MMC cultured with MMC-sensitized T cells for 48 hrs (ADR > T cells), ADR+CQ-treated dormant MMC (ADR+CQ), or ADR+CQ-treated dormant MMC cultured with MMC-sensitized T cells for 48 hrs (ADR+CQ > T cells). Splenic T cells were collected from MMC tumor-bearing FVBN202 mice.
Figure 3 Dormant tumor cells established by ADR or ADR+CQ remain susceptible to immunotherapy The in vitro tumor dormancy was established three weeks after three daily treatments of MMC with ADR or ADR+CQ. Untreated MMC cells were used as control. (A) Apoptosis was determined by FVS viability staining in MMC (control), ADR-treated dormant MMC (ADR), ADR+CQ-treated dormant MMC (ADR+CQ), as well as control MMC cultured with three daily doses of IFN-g and analyzed two days later (50 ng/ml) (IFN-g), ADR-treated dormant MMC cultured with three daily doses of IFN-g (50 ng/ml) and analyzed two days later (ADR > IFN-g), or ADR+CQ-treated dormant MMC cultured with three daily doses of IFN-g (50 ng/ml) and analyzed two days later (ADR+CQ > IFN-g). (B) Apoptosis was determined by FVS viability staining of MMC (control), MMC cultured with MMC-sensitized T cells for 48 hrs (T cells), ADR-treated dormant MMC (ADR), ADR-treated dormant MMC cultured with MMC-sensitized T cells for 48 hrs (ADR > T cells), ADR+CQ-treated dormant MMC (ADR+CQ), or ADR+CQ-treated dormant MMC cultured with MMC-sensitized T cells for 48 hrs (ADR+CQ > T cells). Splenic T cells were collected from MMC tumor-bearing FVBN202 mice.CC BY · PubMed Central
Figure 4 Autophagy knockdown tumor cells become less susceptible to ADR-induced apoptosis Autophagy-deficient MMC (ATG5KD MMC) or autophagy-competent MMC (MMC) were treated with a single dose of ADR alone (1 uM ADR for 2 hrs). Tumor cells were analyzed by Annexin V/PI staining prior to treatment (Day 0) or three days after the treatment (Day 4). Experiments were performed in triplicates.
Figure 4 Autophagy knockdown tumor cells become less susceptible to ADR-induced apoptosis Autophagy-deficient MMC (ATG5KD MMC) or autophagy-competent MMC (MMC) were treated with a single dose of ADR alone (1 uM ADR for 2 hrs). Tumor cells were analyzed by Annexin V/PI staining prior to treatment (Day 0) or three days after the treatment (Day 4). Experiments were performed in triplicates.CC BY · PubMed Central
Figure 5 ADR-induced tumor dormancy in autophagy knockdown tumor cells with polyploid-like morphology compared with autophagy competent tumor cells, in vitro MMC or ATG5KD MMC tumor cells (3 million cells, Day 0) were treated with 3 daily doses of ADR (1uM for 2 hrs), and viable cells were counted at week 3 using trypan blue exclusion. Data represent triplicate experiments (A). Dot plots from each experimental group gated for cell cycle phase based upon DNA content (7-AAD) and Ki-67 expression. Events falling to the left of the G1/G0 gates are considered apoptotic cells (AP). Events falling to the far right of the G2/M gate are considered polyploid-like cells (Poly) (B). Three independent experiments have been performed and data represent 3 replicates ± SEM.
Figure 5 ADR-induced tumor dormancy in autophagy knockdown tumor cells with polyploid-like morphology compared with autophagy competent tumor cells, in vitro MMC or ATG5KD MMC tumor cells (3 million cells, Day 0) were treated with 3 daily doses of ADR (1uM for 2 hrs), and viable cells were counted at week 3 using trypan blue exclusion. Data represent triplicate experiments (A). Dot plots from each experimental group gated for cell cycle phase based upon DNA content (7-AAD) and Ki-67 expression. Events falling to the left of the G1/G0 gates are considered apoptotic cells (AP). Events falling to the far right of the G2/M gate are considered polyploid-like cells (Poly) (B). Three independent experiments have been performed and data represent 3 replicates ± SEM.CC BY · PubMed Central
Figure 6 Earlier relapse of autophagy knockdown tumor cells with polyploid morphology compared with autophagy competent tumor cells, in vivo (A) FVBN202 mice (n = 7) were challenged i.v. with 106 cells ADR-treated dormant control MMC (MMC), or ADR-treated dormant ATG5KD MMC (ATG5KD MMC). Animals were euthanized as soon as they became moribund. Representative tumor relapse in the lung and survival curve are shown. (B) Relapsed tumors were collected and immunohistochemistry slides were prepared by either staining samples with hematoxylin and eosin (H&E) or by Ki67 staining followed by subsequent digitization and analysis with NDP View software (Hamamatsu Photonics). At twenty-times magnification, three representative 0.02 mm2 areas were chosen from the H&E slides containing approximately 100 cells to measure nuclear envelope size. Cells containing a nuclear envelope equal to or greater than 16 um with visible multi-nuclei were considered polyploid-like or high grade cells. The corresponding cell was then analyzed on the Ki67 stained slide to determine Ki67 expression levels. Data was collected from three biological samples. Significance is based on a two-tailed t-test of p < 0.05.
Figure 6 Earlier relapse of autophagy knockdown tumor cells with polyploid morphology compared with autophagy competent tumor cells, in vivo (A) FVBN202 mice (n = 7) were challenged i.v. with 106 cells ADR-treated dormant control MMC (MMC), or ADR-treated dormant ATG5KD MMC (ATG5KD MMC). Animals were euthanized as soon as they became moribund. Representative tumor relapse in the lung and survival curve are shown. (B) Relapsed tumors were collected and immunohistochemistry slides were prepared by either staining samples with hematoxylin and eosin (H&E) or by Ki67 staining followed by subsequent digitization and analysis with NDP View software (Hamamatsu Photonics). At twenty-times magnification, three representative 0.02 mm2 areas were chosen from the H&E slides containing approximately 100 cells to measure nuclear envelope size. Cells containing a nuclear envelope equal to or greater than 16 um with visible multi-nuclei were considered polyploid-like or high grade cells. The corresponding cell was then analyzed on the Ki67 stained slide to determine Ki67 expression levels. Data was collected from three biological samples. Significance is based on a two-tailed t-test of p < 0.05.CC BY · PubMed Central
Figure 7 Autophagy knockdown tumor cells become resistant to T cell-induced apoptosis Neu overexpressing autophagy-deficient MMC (ATG5KD MMC) or autophagy-competent MMC (MMC) were co-cultured with MMC-sensitized T cells and then gated CD45-Neu+ tumor cells were analyzed by Annexin V/PI staining. Data represents triplicate experiments.
Figure 7 Autophagy knockdown tumor cells become resistant to T cell-induced apoptosis Neu overexpressing autophagy-deficient MMC (ATG5KD MMC) or autophagy-competent MMC (MMC) were co-cultured with MMC-sensitized T cells and then gated CD45-Neu+ tumor cells were analyzed by Annexin V/PI staining. Data represents triplicate experiments.CC BY · PubMed Central

Verification

Claims the paper makes

The claims and statistics of this paper haven’t been checked yet.

Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.

A few free checks a day · papers already verified open instantly · results are public

Claims other papers make about this one

Each is checked against this paper’s own text: does it say what the citing paper says it does?

Self-check

Percentages, sums and reported differences re-computed from the paper’s own numbers.

The self-check needs the paper’s full text.

Citations

Cites

Research works printed but not linked: 28

  1. Levine, 2015, Autophagy in malignant transformation and cancer progression, The EMBO journal, 34, 856, 10.15252/embj.201490784 · DOI · Paper page
  2. Gewirtz, 2014, The four faces of autophagy: implications for cancer therapy, Cancer research, 74, 647, 10.1158/0008-5472.CAN-13-2966 · DOI
  3. Yoshimori, 2002, Autophagosome formation in mammalian cells, Cell structure and function, 27, 421, 10.1247/csf.27.421 · DOI
  4. Abdel-Naim, 2014, Chloroquine synergizes sunitinib cytotoxicity via modulating autophagic, apoptotic and angiogenic machineries, Chemico-biological interactions, 217, 28, 10.1016/j.cbi.2014.04.007 · DOI
  5. Shao, 2013, The residual tumor autophagy marker LC3B serves as a prognostic marker in local advanced breast cancer after neoadjuvant chemotherapy, Clinical cancer research, 19, 6853, 10.1158/1078-0432.CCR-13-1617 · DOI
  6. Thorburn, 2012, Chloroquine sensitizes breast cancer cells to chemotherapy independent of autophagy, Autophagy, 8, 200, 10.4161/auto.8.2.18554 · DOI
  7. Evans, 2014, Phase I trial of hydroxychloroquine with dose-intense temozolomide in patients with advanced solid tumors and melanoma, Autophagy, 10, 1369, 10.4161/auto.29118 · DOI
  8. Fornai, 2013, Rapamycin inhibits the growth of glioblastoma, Brain research, 1495, 37, 10.1016/j.brainres.2012.11.044 · DOI
  9. Mariotto, 2017, Metastatic Breast Cancer Cells Enter Into Dormant State and Express Cancer Stem Cells Phenotype Under Chronic Hypoxia, Journal of cellular biochemistry, 118, 3237, 10.1002/jcb.25972 · DOI
  10. Knutson, 2009, Immune-induced epithelial to mesenchymal transitiongenerates breast cancer stem cells, Cancer research, 69, 2887, 10.1158/0008-5472.CAN-08-3343 · DOI
  11. Disis, 2017, Immunization against HIF-1alpha Inhibits the Growth of Basal Mammary Tumors and Targets Mammary Stem Cells, Clinical cancer research, 23, 3396, 10.1158/1078-0432.CCR-16-1678 · DOI
  12. Kinsella, 2011, Impact of Autophagy on Chemotherapy and Radiotherapy Mediated Tumor Cytotoxicity: “To Live or not to Live”, Frontiers in oncology, 1, 30
  13. Kroemer, 2011, Association and dissociation of autophagy, apoptosis and necrosis by systematic chemical study, Oncogene, 30, 4544, 10.1038/onc.2011.168 · DOI
  14. Aguirre-Ghiso, 2013, Regulation of tumor cell dormancy by tissue microenvironments and autophagy, Advances in Experimental Medicine and Biology, 734, 73, 10.1007/978-1-4614-1445-2_5 · DOI
  15. Manjili, 2016, Tumor-reactive immune cells protect against metastatic tumor and induce immunoediting of indolent but not quiescent tumor cells, J Leukoc Biol, 100, 625, 10.1189/jlb.5A1215-580R · DOI
  16. Chang, 2016, The autophagy inhibitor chloroquine targets cancer stem cells in triple negative breast cancer by inducing mitochondrial damage and impairing DNA break repair, Cancer letters, 376, 249, 10.1016/j.canlet.2016.04.002 · DOI
  17. Dave, 2014, Chloroquine eliminates cancer stem cells through deregulation of Jak2 and DNMT1, Stem cells (Dayton, Ohio), 32, 2309, 10.1002/stem.1746 · DOI
  18. White, 2007, Autophagy mitigates metabolic stress and genome damage in mammary tumorigenesis, Genes & development, 21, 1621, 10.1101/gad.1565707 · DOI
  19. White, 2007, Autophagy suppresses tumor progression by limiting chromosomal instability, Genes & development, 21, 1367, 10.1101/gad.1545107 · DOI
  20. Klionsky, 2008, Autophagy fights disease through cellular self-digestion, Nature, 451, 1069, 10.1038/nature06639 · DOI · Paper page
  21. Levine, 1999, Induction of autophagy and inhibition of tumorigenesis by beclin 1, Nature, 402, 672, 10.1038/45257 · DOI · Paper page
  22. Spielmann, 2015, Combined evaluation of LC3B puncta and HMGB1 expression predicts residual risk of relapse after adjuvant chemotherapy in breast cancer, Autophagy, 11, 1878, 10.1080/15548627.2015.1082022 · DOI
  23. Shao, 2015, Expression of autophagy-related proteins ATG5 and FIP200 predicts favorable disease-free survival in patients with breast cancer, Biochemical and biophysical research communications, 458, 816, 10.1016/j.bbrc.2015.02.037 · DOI
  24. Wang, 2017, CISD2 promotes the proliferation of glioma cells via suppressing beclin1mediated autophagy and is targeted by microRNA449a, Molecular medicine reports, 16, 7939, 10.3892/mmr.2017.7642 · DOI
  25. Manjili, 2007, HER-2/neu antigen loss and relapse of mammary carcinoma are actively induced by T cell-mediated anti-tumor immune responses, European journal of immunology, 37, 675, 10.1002/eji.200636639 · DOI
  26. Muller, 1992, Expression of the neu protooncogene in the mammary epithelium of transgenic mice induces metastatic disease, Proceedings of the National Academy of Sciences of the United States of America, 89, 10578, 10.1073/pnas.89.22.10578 · DOI
  27. Manjili, 2008, Danger signals and nonself entity of tumor antigen are both required for eliciting effective immune responses against HER-2/neu positive mammary carcinoma: implications for vaccine design, Cancer Immunol Immunother, 57, 1391, 10.1007/s00262-008-0475-8 · DOI
  28. Manjili, 2011, Activated NKT cells and NK cells render T cells resistant to myeloid-derived suppressor cells and result in an effective adoptive cellular therapy against breast cancer in the FVBN202 transgenic mouse, J Immunol, 187, 708, 10.4049/jimmunol.1100502 · DOI

Cited by

Sorted by citations

See all 76

Metadata

Authors
Aqbi, H. F., Tyutyunyk-Massey, L., Keim, R. C., Butler, S. E., Thekkudan, T., Joshi, S., Smith, T. M., Bandyopadhyay, D., Idowu, M. O., Bear, H. D., Payne, K. K., Gewirtz, D. A., & Manjili, M. H.
Affiliations
  • Department of Microbiology and Immunology, Virginia Commonwealth University School of Medicine, Richmond, 23298, VA, USA
  • Department of Pharmacology and Toxicology, Virginia Commonwealth University School of Medicine, Richmond, 23298, VA, USA
  • Department of Human and Molecular Genetics, Virginia Commonwealth University School of Medicine, Richmond, 23298, VA, USA
  • Massey Cancer Center, Virginia Commonwealth University School of Medicine, Richmond, 23298, VA, USA
  • Department of Immunology, Moffitt Cancer Center, Tampa, 33612, FL, USA
Venue
Oncotarget
Publisher
Impact Journals, LLC
Published
2018-04-24
Volume
9
Issue
31
Pages
22113-22122
ISSN
1949-2553
DOI
10.18632/oncotarget.25197
PMC
PMC5955162
Cited by
76
References
2
Language
en

About these checks

Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.

Statistics and arithmetic are recomputed, not judged.

Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.

What counts as a claim ›How we measure confidence ›How we check this ›

Are you an author, or do you see a problem with this page? Request a review