Journal: Oncology reports

Article Title: Radioresistant breast cancer cells exhibit increased resistance to chemotherapy and enhanced invasive properties due to cancer stem cells.

doi: 10.3892/or.2018.6714

Figure Lengend Snippet: Figure 2. RT‑R‑MDA‑MB‑231 cells demonstrate higher levels of CSCs marker CD44, but lower levels of CD24 compared with MDA‑MB‑231 cells. Classical CSCs markers CD44 and CD24 were detected in the low metastatic breast cancer cells MCF‑7 and RT‑R‑MCF‑7 (A) and highly metastatic breast cancer cells MDA‑MB‑231 and RT‑R‑MDA‑MB‑231 (B) using the specific antibodies by western blotting. Data represent mean values ± standard error of the mean of three independent experiments in triplicate. **P<0.01 compared with control. CD, cluster of differentiation; CSCs, cancer stem cells; RT‑R, radioresistant.

Article Snippet: Isolation of CD24low/CD44high breast cancer stem cells was performed using a MagCollect CD24- CD44+ Breast Cancer Stem Cell Isolation kit (R&D Systems, Inc., Minneapolis, MN, USA) following the manufacturer's protocol.

Techniques: Marker, Western Blot, Control

Journal: Oncology reports

Article Title: Radioresistant breast cancer cells exhibit increased resistance to chemotherapy and enhanced invasive properties due to cancer stem cells.

doi: 10.3892/or.2018.6714

Figure Lengend Snippet: Figure 3. RT‑R‑MDA‑MB‑231 cells increase the populations of CD24low/CD44high cells, and expression levels of other CSCs markers Notch‑4, Oct‑3/4 and ALDH1. (A) CD24low/CD44high cells were isolated from MDA‑MB‑231 and RT‑R‑MDA‑MB cells, and then the number of CD24low/CD44high breast cancer stem cells was quantified by flow cytometry. (B) MDA‑MB‑231 and RT‑R‑MDA‑MB‑231 were labeled with anti‑CD24 and anti‑CD44 antibodies, and the percentages of CD24 or CD44‑expressed subpopulation in cells were determined by flow cytometry. (C) Other cancer stem cell markers, including Notch‑4, Oct3/4 and ALDH1 were detected in the MDA‑MB‑231 and RT‑R‑MDA‑MB‑231 cells using the specific antibodies by western blotting. Data represent mean values ± standard error of the mean of three independent experiments in triplicate. *P<0.05 and **P<0.01 compared with MDA‑MB‑231 cells. ALDH1, aldehyde dehydrogenase 1; CD, cluster of differentiation; CSCs, cancer stem cells; Oct, octamer‑binding transcription factor; RT‑R, radioresistant.

Article Snippet: Isolation of CD24low/CD44high breast cancer stem cells was performed using a MagCollect CD24- CD44+ Breast Cancer Stem Cell Isolation kit (R&D Systems, Inc., Minneapolis, MN, USA) following the manufacturer's protocol.

Techniques: Expressing, Isolation, Flow Cytometry, Labeling, Western Blot

Journal: Nature Communications

Article Title: An in-situ peptide-antibody self-assembly to block CD47 and CD24 signaling enhances macrophage-mediated phagocytosis and anti-tumor immune responses

doi: 10.1038/s41467-024-49825-6

Figure Lengend Snippet: a Schematic illustration of tumor immune quiescent microenvironment, in situ assembly of PAC-SABIs on the cancer cell surface, and blockage of CD47 and CD24 phagocytic checkpoints by PAC-SABIs. Figure was created with BioRender.com and released under a Creative Commons Attribution-NonCommercial-NoDerivs 4.0 international license. b Time-dependent CLSM images of 4T1 cells treated with NBD-labeled PAC-SABIs. Scale bar: 20 μm. Three independent experiments were performed. c Time-dependent CLSM images of PAN02 cells treated with NBD-labeled PAC-SABIs. Scale bar: 20 μm. Three independent experiments were performed. d Merged bright field and fluorescence images of 4T1 cell treated with NBD-labeled PAC-SABIs for 120 min. Scale bar: 20 μm. Three independent experiments were performed. e CLSM images of 4T1 cell treated with Dil dye (red) and NBD-labeled PAC-SABIs for 120 min. Scale bar: 10 μm. Three independent experiments were performed. f , g Fluorescence distribution of NBD-labeled PAC-SABIs on 4T1 cell. Three independent experiments were performed. h Time-dependent SEM images of 4T1 cells treated with PAC-SABIs. The red arrows point to PAC-SABIs on cell membrane. Scale bar: 2 μm. Three independent experiments were performed. i Time-dependent SEM images of PAN02 cells treated with PAC-SABIs. The red arrows point to the PAC-SABIs on cell membrane. Scale bar: 2 μm. Three independent experiments were performed. j CLSM images of 3D 4T1 spheroids treated with Cy5.5 PAC-SABIs, Calcein-AM, and Hoechst 33342. Scale bar: 50 μm. Three independent experiments were performed. k CLSM images of 3D 4T1 spheroids along the z -axis position. Scale bar: 50 μm. Three independent experiments were performed.

Article Snippet: Treatment: anti-mouse CD24 antibody (BioXcell, #BE0360, clone M1/69), anti-human CD24 antibody (Novus Biologics, #NB100-64861, clone SN3), anti-mouse CD47 antibody (BioXcell, #BE0270, clone MIAP301), anti-mouse PD-1 antibody (BioXcell, #BE0273, clone 29F.1A12™), anti-CSF1R antibody (BioXcell, #BE0213, clone AFS98).

Techniques: In Situ, Labeling, Fluorescence, Membrane

Journal: Nature Communications

Article Title: An in-situ peptide-antibody self-assembly to block CD47 and CD24 signaling enhances macrophage-mediated phagocytosis and anti-tumor immune responses

doi: 10.1038/s41467-024-49825-6

Figure Lengend Snippet: a Phagocytosis images of pHrodo-red + over time. Scale bar: 50 μm. Three independent experiments were performed. b Phagocytosis of 4T1 cells, in the presence of IgG control or PAC-SABIs. The error bars represent the mean ± SD ( n = 3 independent experiments). c Representative 3D CLSM image reconstruction of in vitro phagocytosis of 4T1 cells (pHrodo-red + , red) by BMDMs and RAW264.7 cells (Calcein-AM, green). Three independent experiments were performed. d Representative flow cytometry plots depicting BMDM phagocytosis of 4T1 cells treated with IgG control, anti-CD24 mAb, SAMIs and PAC-SABIs. e Quantitative analysis of BMDM flow cytometry results. The error bars represent the mean ± SD ( n = 3 independent experiments; ** p < 0.01, *** p < 0.001; the p value was analyzed by a two-tailed unpaired Student’s t test). f Representative flow cytometry plots depicting RAW264.7 cell phagocytosis of 4T1 cells treated with IgG control, anti-CD24 mAb, SAMIs and PAC-SABIs. g Quantitative analysis of RAW264.7 cell flow cytometry results. The error bars represent the mean ± SD ( n = 3 independent experiments; ** p < 0.01; the p value was analyzed by a two-tailed unpaired Student’s t test). Source data are provided as a Source Data file.

Article Snippet: Treatment: anti-mouse CD24 antibody (BioXcell, #BE0360, clone M1/69), anti-human CD24 antibody (Novus Biologics, #NB100-64861, clone SN3), anti-mouse CD47 antibody (BioXcell, #BE0270, clone MIAP301), anti-mouse PD-1 antibody (BioXcell, #BE0273, clone 29F.1A12™), anti-CSF1R antibody (BioXcell, #BE0213, clone AFS98).

Techniques: Control, In Vitro, Flow Cytometry, Two Tailed Test

Journal: Cells

Article Title: Chronic High-Salt Diet Activates Tumor-Initiating Stem Cells Leading to Breast Cancer Proliferation

doi: 10.3390/cells13110912

Figure Lengend Snippet: Enumeration of TISC frequency in various murine passages following dietary salt modification. The frequency of CD44+CD24−TISCs cells in the isolated tumor cells among RS (grey) and HS (red) diet cohorts was enumerated by flow cytometry. ( A , B ) Representative flow cytometry plot of TISCs from passage 1 and passage 4 of Py230-C57Bl/6J HS diet cohort. ( C , D ) Changes in TISC frequency with each passage in Py230-C57Bl/6J ( C ) and 4T1-BALB/cJ ( D ) murine tumor models. ( E – N ) The mRNA expression of TISC markers Cadherin 1 ( E , J ), Snail2 ( F , K ), Aldh1A1 ( G , L ), Sox2 ( H , M ), and ITGA6 ( I , N ) in four passages of Py230-C57Bl/6J and 4T1-BALB/cJ tumor models (respectively). Data analyzed by one-way ANOVA for multiple comparisons and presented as mean ± SEM; n = 8 (biological replicates) per cohort; (*) p -value < 0.05.

Article Snippet: The TISCs were isolated from murine tumors by CD24 − CD44 + breast tumor stem cells using immunomagnetic beads as per the manufacturer’s specifications and reagents (catalog#MAGH111, R&D Systems, Minneapolis, MN, USA).

Techniques: Modification, Isolation, Flow Cytometry, Expressing

Journal: Cells

Article Title: Chronic High-Salt Diet Activates Tumor-Initiating Stem Cells Leading to Breast Cancer Proliferation

doi: 10.3390/cells13110912

Figure Lengend Snippet: TGFβ signaling induced the expression of immune-inhibitory CD80 on TISCs. ( A ) Representative flow cytometry histogram of TGFβR2 expression in passages 1 (blue), 2 (pink), 3 (olive green), and 4 (purple) from the HS diet cohort of the Py230-C57Bl/6J tumor model. ( B ) Comparison of the relative expression of TGFβR2 in each of the four passages on CD4+CD24-TISCs isolated from RS (grey) and HS (red) diet cohorts of the Py230-C57Bl/6J tumor model. ( C ) Representative flow cytometry histogram of pSMAD2/3 intracellular expression in each of the four passages from the HS diet cohort of the Py230-C57Bl/6J tumor model. ( D ) Comparison of the relative expression of pSMAD2/3 in each of the four passages on CD4+CD24-TISCs isolated from the RS and HS diet cohorts of the Py230-C57Bl/6J tumor model. ( E ) Representative flow cytometry histogram of CD80 surface expression in each of the four passages from the HS diet cohort of the Py230-C57Bl/6J tumor model. ( F ) Comparison of the relative expression of CD80 in each of the four passages on CD4+CD24-TISCs isolated from the RS and HS diet cohorts of the Py230-C57Bl/6J tumor model. ( G ) Representative flow cytometry plot to determine the TGFβR2/CD80 double-positive TISCs in passage 4 of the HS diet cohort from the Py230-C57Bl/6J tumor model. ( H ) Comparison of the relative expression of TGFβR2/CD80 double-positive TISCs in each of the four passages on CD4+CD24-TISCs isolated from the RS and HS diet cohorts of the Py230-C57Bl/6J tumor model. ( I ) Comparison of the relative expression of CD80 following TGFβ (80 ng/mL) stimulation for 5 days on CD4+CD24-TISCs isolated from each of the four passages of the RS and HS diet cohorts in the Py230-C57Bl/6J tumor model. ( J – M ) Comparison of the relative expression of TGFβR2 ( J ), pSMAd2/3 ( K ), CD80 ( L ), and TGFβR2/CD80 double-positive cells ( M ) in each of the four passages on CD4+CD24-TISCs isolated from the RS (grey) and HS (red) diet cohorts of the 4T1-BALB/cJ tumor model. ( N ) Comparison of the relative expression of CD80 following TGFβ (80 ng/mL) treatment for 5 days on CD4+CD24-TISCs isolated from each of the four passages of the RS and HS diet cohorts of the 4T1-BALB/cJ tumor model. Data analyzed by one-way ANOVA for multiple comparisons and presented as mean ± SEM; n = 8 (biological replicates) per cohort; (*) p -value < 0.05.

Article Snippet: The TISCs were isolated from murine tumors by CD24 − CD44 + breast tumor stem cells using immunomagnetic beads as per the manufacturer’s specifications and reagents (catalog#MAGH111, R&D Systems, Minneapolis, MN, USA).

Techniques: Expressing, Flow Cytometry, Comparison, Isolation

Journal: International Journal of Oncology

Article Title: CD44v3 + /CD24 − cells possess cancer stem cell-like properties in human oral squamous cell carcinoma

doi: 10.3892/ijo.2015.3261

Figure Lengend Snippet: Expression of CD44v3/CD24 in SAS and OSC20. SAS and OSC20 are labeled with CD44v3 and CD24, and then analyzed by FCM. In SAS, the proportion of CD44v3 + /CD24 − , CD44v3 + /CD24 + , CD44v3 − /CD24 − and CD44v3 − /CD24 + cells is 10.7, 48.2, 7.8 and 33.3%, respectively. Whereas, the proportion of CD44v3 + /CD24 − , CD44v3 + /CD24 + , CD44v3 − /CD24 − and CD44v3 − /CD24 + cells is 24.1, 52.0, 11.2 and 12.7%, respectively.

Article Snippet: Although the CD44v3 + /CD24 − cell fraction in SAS or OSC20 did not show a higher proliferative ability than each of the other fractions (CD44v3 − /CD24 + , CD44v3 − /CD24 − and CD44v3 − /CD24 + cells), the CD44v3 + /CD24 − cell fraction showed significantly higher sphere forming ability, a higher CDDP or 5-FU resistance, suggesting that the CD44v3 + /CD24 − cell fraction had CSC-LC properties in SAS or OSC20.

Techniques: Expressing, Labeling

Journal: International Journal of Oncology

Article Title: CD44v3 + /CD24 − cells possess cancer stem cell-like properties in human oral squamous cell carcinoma

doi: 10.3892/ijo.2015.3261

Figure Lengend Snippet: Biological features of sorted cell fractions in SAS and OSC20 in vitro . (A) CD44v3 + /CD24 − cells in SAS show a higher proliferative ability than that of CD44v3 − /CD24 − and CD44v3 − /CD24 + cells. CD44v3 + /CD24 − cells in OSC20 show a higher proliferative ability than that of CD44v3 − /CD24 − cells ( * P<0.05). (B) The sphere forming ability of CD44v3 + /CD24 − cells in both SAS and OSC20 is significantly higher than in the other fractions ( * P<0.05, *** P<0.001). The sphere forming ability of CD44v3 + /CD24 + cells is also higher than that of CD44v3 − /CD24 − and CD44v3 − /CD24 + cells ( *** P<0.001). (C) In SAS, the viability of CD44v3 + /CD24 − cells is significantly higher than the other fractions after treatment with 1 μM CDDP, or treatment with 10 or 100 μM of 5-FU and was significantly higher than that of CD44v3 + /CD24 + or CD44v3 − /CD24 + cells after treatment with 5 μM CDDP, or treatment with 100 or 1,000 nM cetuximab ( * P<0.05, ** P<0.01, *** P<0.001). In OSC20, the viability of CD44v3 + /CD24 − cells is significantly higher than that of CD44v3 + /CD24 + or CD44v3 − /CD24 + cells after treatment with 1 or 5 μM CDDP ( *** P<0.001).

Article Snippet: Although the CD44v3 + /CD24 − cell fraction in SAS or OSC20 did not show a higher proliferative ability than each of the other fractions (CD44v3 − /CD24 + , CD44v3 − /CD24 − and CD44v3 − /CD24 + cells), the CD44v3 + /CD24 − cell fraction showed significantly higher sphere forming ability, a higher CDDP or 5-FU resistance, suggesting that the CD44v3 + /CD24 − cell fraction had CSC-LC properties in SAS or OSC20.

Techniques: In Vitro

Journal: International Journal of Oncology

Article Title: CD44v3 + /CD24 − cells possess cancer stem cell-like properties in human oral squamous cell carcinoma

doi: 10.3892/ijo.2015.3261

Figure Lengend Snippet: Quantification of mRNA expression of CSC-LC property-related genes in sorted cell fractions by real-time PCR. In SAS, CD44v3 + /CD24 − cells show significantly higher mRNA expression of transporter-related genes (ABCB1 and ABCG2), ALDH1A1, anti-apoptotic genes (BCL2) and self-replication genes (Oct-4 and Nanog) than the other fractions. The experiments were repeated at least three times for each cell line, and almost identical results were obtained ( * P<0.05, ** P<0.01, *** P<0.001).

Article Snippet: Although the CD44v3 + /CD24 − cell fraction in SAS or OSC20 did not show a higher proliferative ability than each of the other fractions (CD44v3 − /CD24 + , CD44v3 − /CD24 − and CD44v3 − /CD24 + cells), the CD44v3 + /CD24 − cell fraction showed significantly higher sphere forming ability, a higher CDDP or 5-FU resistance, suggesting that the CD44v3 + /CD24 − cell fraction had CSC-LC properties in SAS or OSC20.

Techniques: Expressing, Real-time Polymerase Chain Reaction

Journal: International Journal of Oncology

Article Title: CD44v3 + /CD24 − cells possess cancer stem cell-like properties in human oral squamous cell carcinoma

doi: 10.3892/ijo.2015.3261

Figure Lengend Snippet: Tumorigenicity of sorted cell fractions in SAS.

Article Snippet: Although the CD44v3 + /CD24 − cell fraction in SAS or OSC20 did not show a higher proliferative ability than each of the other fractions (CD44v3 − /CD24 + , CD44v3 − /CD24 − and CD44v3 − /CD24 + cells), the CD44v3 + /CD24 − cell fraction showed significantly higher sphere forming ability, a higher CDDP or 5-FU resistance, suggesting that the CD44v3 + /CD24 − cell fraction had CSC-LC properties in SAS or OSC20.

Techniques:

Journal: International Journal of Oncology

Article Title: CD44v3 + /CD24 − cells possess cancer stem cell-like properties in human oral squamous cell carcinoma

doi: 10.3892/ijo.2015.3261

Figure Lengend Snippet: Immunostaining of (A–D) CD44v3 and (E–H) CD24 in OSCC tissues. The staining intensity was graded into 4 levels, i.e., 0, negative; 1, weakly positive; 2, moderately positive; 3, strongly positive.

Article Snippet: Although the CD44v3 + /CD24 − cell fraction in SAS or OSC20 did not show a higher proliferative ability than each of the other fractions (CD44v3 − /CD24 + , CD44v3 − /CD24 − and CD44v3 − /CD24 + cells), the CD44v3 + /CD24 − cell fraction showed significantly higher sphere forming ability, a higher CDDP or 5-FU resistance, suggesting that the CD44v3 + /CD24 − cell fraction had CSC-LC properties in SAS or OSC20.

Techniques: Immunostaining, Staining

Journal: International Journal of Oncology

Article Title: CD44v3 + /CD24 − cells possess cancer stem cell-like properties in human oral squamous cell carcinoma

doi: 10.3892/ijo.2015.3261

Figure Lengend Snippet: Immunostaining of CD44v3 and CD24 in the non-invasive portion and invasive portion. Box and Whisker plots of CD44v3 and CD24 immunoscoring of the non-invasive portion and invasive portion showing median (bold line) and 25th to 75th percentiles ( ** P<0.01).

Article Snippet: Although the CD44v3 + /CD24 − cell fraction in SAS or OSC20 did not show a higher proliferative ability than each of the other fractions (CD44v3 − /CD24 + , CD44v3 − /CD24 − and CD44v3 − /CD24 + cells), the CD44v3 + /CD24 − cell fraction showed significantly higher sphere forming ability, a higher CDDP or 5-FU resistance, suggesting that the CD44v3 + /CD24 − cell fraction had CSC-LC properties in SAS or OSC20.

Techniques: Immunostaining, Whisker Assay

Journal: International Journal of Oncology

Article Title: CD44v3 + /CD24 − cells possess cancer stem cell-like properties in human oral squamous cell carcinoma

doi: 10.3892/ijo.2015.3261

Figure Lengend Snippet: Association of CD44v3 and CD24 immunoexpression with clinicopathological parameters in 50 OSCC patients.

Article Snippet: Although the CD44v3 + /CD24 − cell fraction in SAS or OSC20 did not show a higher proliferative ability than each of the other fractions (CD44v3 − /CD24 + , CD44v3 − /CD24 − and CD44v3 − /CD24 + cells), the CD44v3 + /CD24 − cell fraction showed significantly higher sphere forming ability, a higher CDDP or 5-FU resistance, suggesting that the CD44v3 + /CD24 − cell fraction had CSC-LC properties in SAS or OSC20.

Techniques:

Journal: International Journal of Oncology

Article Title: CD44v3 + /CD24 − cells possess cancer stem cell-like properties in human oral squamous cell carcinoma

doi: 10.3892/ijo.2015.3261

Figure Lengend Snippet: Association of CD44v3/CD24 immunophenotype with clinicopathological parameters in 50 OSCC patients.

Article Snippet: Although the CD44v3 + /CD24 − cell fraction in SAS or OSC20 did not show a higher proliferative ability than each of the other fractions (CD44v3 − /CD24 + , CD44v3 − /CD24 − and CD44v3 − /CD24 + cells), the CD44v3 + /CD24 − cell fraction showed significantly higher sphere forming ability, a higher CDDP or 5-FU resistance, suggesting that the CD44v3 + /CD24 − cell fraction had CSC-LC properties in SAS or OSC20.

Techniques:

Journal: International Journal of Oncology

Article Title: CD44v3 + /CD24 − cells possess cancer stem cell-like properties in human oral squamous cell carcinoma

doi: 10.3892/ijo.2015.3261

Figure Lengend Snippet: Overall survival (OS) rates and CD44v3/CD24 immunophenotypes in the invasive portion. (A) CD44v3 + cases tended to show poor OS compared with CD44v3 − cases (P=0.055). (B) There was no significant difference in OS between CD24 + cases and CD24 − cases (P=0.585). (C) The Kaplan Meier curves for OS in OSCC patients were subdivided into 4 groups according to CD44v3/CD24 immunophenotypes (CD44v3 + /CD24 − , CD44v3 + /CD24 − , CD44v3 − /CD24 − and CD44v3 − /CD24 + ). CD44v3 + /CD24 − cases showed poor OS, and there was a significant difference between CD44v3 + /CD24 − and CD44v3 − /CD24 − cases (P=0.029).

Article Snippet: Although the CD44v3 + /CD24 − cell fraction in SAS or OSC20 did not show a higher proliferative ability than each of the other fractions (CD44v3 − /CD24 + , CD44v3 − /CD24 − and CD44v3 − /CD24 + cells), the CD44v3 + /CD24 − cell fraction showed significantly higher sphere forming ability, a higher CDDP or 5-FU resistance, suggesting that the CD44v3 + /CD24 − cell fraction had CSC-LC properties in SAS or OSC20.

Techniques: