Journal: Cell Genomics

Article Title: Defining hypoxia in cancer: A landmark evaluation of hypoxia gene expression signatures

doi: 10.1016/j.xgen.2025.100764

Figure Lengend Snippet: Performance of the Tardon signature using the IQM in scRNA-seq data for MCF7 and HCC1806 cells under normoxic and hypoxic conditions After reaching maximum accuracy in bulk RNA-seq, Tardon performs extremely well in the more sparse scRNA-seq data. This is shown in all three panels. (A) 3D uniform manifold approximation and projection (UMAP) representation with x and y axes displaying UMAP dimensions and z axis showing the IQM score of the Tardon signature for individual cells. Separation between normoxia and hypoxia is seen in both cell lines. (B) IQM scores across all tested signatures, annotated vertically by experimental condition and horizontally by significance level. The heatmap reveals high concordance between single-cell and bulk data. Gray vertical lines indicate instances where insufficient genes are available for IQM calculation, which requires a minimum of four values.

Article Snippet: HCC1806 , Cytion , RRID: CVCL_1258 Cat# 300467-1VL.

Techniques: RNA Sequencing

Journal: Cell Genomics

Article Title: Defining hypoxia in cancer: A landmark evaluation of hypoxia gene expression signatures

doi: 10.1016/j.xgen.2025.100764

Figure Lengend Snippet:

Article Snippet: HCC1806 , Cytion , RRID: CVCL_1258 Cat# 300467-1VL.

Techniques: Gene Expression, Software, Sequencing, Sample Prep, Recombinant, Saline, Reverse Transcription

Journal: Advanced Science

Article Title: YAP Dictates Mitochondrial Redox Homeostasis to Facilitate Obesity‐Associated Breast Cancer Progression

doi: 10.1002/advs.202103687

Figure Lengend Snippet: Adipocytes promote aggressiveness of TNBC in a YAP‐driven antioxidant manner. A) BODIPY staining of lipid accumulation in Py8119 cells incubated with lean‐CM or obese‐CM for 24 h. B) Western blot analysis of total and phosphorylated YAP levels in Py8119 cells treated with lean‐CM or obese‐CM. C) Western blot analysis of total and phosphorylated YAP levels in the whole‐cell lysate (WCL) and total YAP levels in the nuclear fraction (NF) of Py8119 cells treated with different concentrations of adi‐CM. D) Py8119 cells were pretreated with etomoxir, rotenone, antimycin, or oligomycin and then incubated with adi‐CM for 24 h. Expressions of YAP and phosphorylated YAP were assessed by Western blotting. E) qPCR analysis of MSRA , GCLC , GSR , and PRDX1 gene expressions in Py8119/shLacZ and shYAP cells exposed to adi‐CM. (F‐H) Py8119/shLacZ and shYAP cells were pretreated with NAC (2 × 10 −3 m ) or MitoTEMPO (5 × 10 −6 m ) followed by incubation with adi‐CM for another 24 h, F) DCFH‐DA+ and MitoSOX+ cells were analyzed by flow cytometry, G) fluorescent microscopic observations of DCFH‐DA, MitoSOX, and BODIPY 581/591 C11 staining, and H) cell proliferation and transwell migration and invasion assays were performed. I) MDA‐MB‐231/shLacZ, HCC1806/shLacZ, MDA‐MB‐231/shYAP, and HCC1806/shYAP cells were pretreated with NAC or MitoTEMPO followed by incubation with adi‐CM for 24 h. Cell proliferation and transwell migration and invasion assays were performed. Data are presented as the mean ± SD. * p < 0.05, ** p < 0.01, *** p < 0.001, as determined by an unpaired two‐tailed Student's t ‐test.

Article Snippet: Human MDA‐MB‐231 and HCC1806 TNBC cell lines, the mouse Py8119 TNBC cell line, and mouse 3T3‐L1 preadipocyte cells were purchased from the Bioresource Collection Research Center (Hsinchu, Taiwan) or ATCC.

Techniques: Staining, Incubation, Western Blot, Flow Cytometry, Migration, Two Tailed Test

Journal: Pharmaceutics

Article Title: Synthesis and Evaluation of Small Molecule Drug Conjugates Harnessing Thioester-Linked Maytansinoids

doi: 10.3390/pharmaceutics14071316

Figure Lengend Snippet: Cytotoxic effects on human triple-negative breast cancer HCC1806 cells. With a 72 h incubation of conjugates Zn8_DM1 , Zn10_DM4 , cytotoxic maytansinoids DM1 , and DM4 with the cells, reduction of tetrazolium by viable cells was determined.

Article Snippet: Human triple-negative breast cancer HCC1806 (1 × 10 6 cells) cells were inoculated subcutaneously into the left flank of 6-week-old female nude mice (BioLASCO Taiwan, Taipei, Taiwan) using a 1 mL syringe with a 24G needle (AN-2425R, Terumo Medical Corp., Tokyo, Japan).

Techniques: Incubation

Journal: Pharmaceutics

Article Title: Synthesis and Evaluation of Small Molecule Drug Conjugates Harnessing Thioester-Linked Maytansinoids

doi: 10.3390/pharmaceutics14071316

Figure Lengend Snippet: ( A ) In vitro SPR PS-association studies. Sensorgrams are generated using a Biacore T200. ZnDPA , Zn8_DM1 , Zn10_DM4 , and DM1 were analyzed across a two-fold serial concentration dilution. The association of liposome (DOPC/ DOPS (3:1, v / v )) immobilized on an L1 chip at 5000 RU was monitored for 1 min and the dissociation time was 1 min. ( B ) In vivo targeting ability of ZnDPA and detection of PS-expression in the HCC1806 tumor xenograft model. Representative IVIS images of fluorescence probe PSVue794 and Dye_794 in mice at the indicated time-point with a single intravenous dose of 2 mg/kg.

Article Snippet: Human triple-negative breast cancer HCC1806 (1 × 10 6 cells) cells were inoculated subcutaneously into the left flank of 6-week-old female nude mice (BioLASCO Taiwan, Taipei, Taiwan) using a 1 mL syringe with a 24G needle (AN-2425R, Terumo Medical Corp., Tokyo, Japan).

Techniques: In Vitro, Generated, Concentration Assay, In Vivo, Expressing, Fluorescence

Journal: Pharmaceutics

Article Title: Synthesis and Evaluation of Small Molecule Drug Conjugates Harnessing Thioester-Linked Maytansinoids

doi: 10.3390/pharmaceutics14071316

Figure Lengend Snippet: In vivo anti-HCC1806 tumor efficacies. The treatment regimen is shown in mg/kg, and dosing frequency per week is indicated by red arrows. The amount of cytotoxic maytansinoids DM1 and DM4 deployed was calculated from the percentage of DM1 and DM4 in the total dose of conjugates Zn8_DM1 and Zn10_DM4 used. ( A ) Comparisons of anti-HCC1806 triple-negative breast cancer activities and body weight changes between Zn8_DM1 , Zn10_DM4 , cytotoxic maytansinoids DM1 and DM4 when administered intravenously at indicated time-point. ( B ) Treatment of large (~900 mm 3 ) HCC1806 triple-negative breast cancer tumor and body weight changes with weekly doses of conjugates Zn8_DM1 and Zn10_DM4 at 2 mg/kg, * p < 0.05 via unpaired t -test.

Article Snippet: Human triple-negative breast cancer HCC1806 (1 × 10 6 cells) cells were inoculated subcutaneously into the left flank of 6-week-old female nude mice (BioLASCO Taiwan, Taipei, Taiwan) using a 1 mL syringe with a 24G needle (AN-2425R, Terumo Medical Corp., Tokyo, Japan).

Techniques: In Vivo

Journal: International Journal of Biological Sciences

Article Title: Targeting the KLF5-EphA2 axis can restrain cancer stemness and overcome chemoresistance in basal-like breast cancer

doi: 10.7150/ijbs.82567

Figure Lengend Snippet: EphA2 is highly expressed in BLBC and promotes breast cancer cell stemness. (A) According to database analysis (bc-GenExMiner v4.9), EphA2 mRNA levels were found to be highly expressed in basal-like breast cancers (BLBC); (B) According to TCGA database analysis, EphA2 mRNA levels were found to be highly expressed in BLBC patients; (C-I) After knockdown of EphA2 in HCC1937 and HCC1806 (C), flow detection of ALDH + (D-E) and CD44 + /CD24 - (F-G) cell ratios and mamosphere formation ratio was performed (H-I), Scale bar=250 μm; (J-K) Dilution tumorigenesis assay in nude mice (J), tumorigenesis rate statistics table (K).

Article Snippet: HCC1806 cells (1×10 6 ) were resuspended in Matrigel (#354234, Corning, BD Biocoat; 1:1 diluted with RPMI-1640 serum-free medium) and injected into the fourth pair of mammary gland fat pads.

Techniques: Knockdown

Journal: International Journal of Biological Sciences

Article Title: Targeting the KLF5-EphA2 axis can restrain cancer stemness and overcome chemoresistance in basal-like breast cancer

doi: 10.7150/ijbs.82567

Figure Lengend Snippet: KLF5 positively regulates EphA2 expression in BLBC. (A) Knockdown of KLF5 in HCC1937 and HCC1806 cells for RNA-seq and 33 genes were found to be positively regulated by KLF5, including EphA2. (B-C) Knockdown of KLF5 was confirmed to suppress EphA2 expression by RT‒qPCR and WB assay; (D) Overexpression of KLF5 promotes EphA2 expression by WB assay; (E) ChIP-seq results of KLF5 in HCC1937 and HCC1806 cells showed two potential binding sites of KLF5 in the EphA2 gene locus, which were named P1 and P2, respectively; (F) Validation of KLF5 binding sites P1 and P2 in the EphA2 gene by ChIP‒qPCR in HCC1937 and HCC1806, respectively. P (Positive) or N (Negative) is the primer targeting P1/P2 or distant non-peak sequence, respectively. G . JASPAR database analysis of the KLF5 binding motif and potential binding sites of KLF5 in the promoter region of EphA2 (Site 3 is out of P1/P2 and Sites 1/2 share common motif); (H) Luciferase activity assay for detecting the transcriptional activity of WT and mutated promoters of EphA2 in HEK293T cells in the absence or presence of KLF5 overexpression. (I) In multiple breast cancer cell lines and normal breast epithelial cell lines, a WB assay found a certain trend of coexpression of KLF5 and EphA2; (J) Database (bc-GenExMiner v4.9) analysis revealed a correlation between the mRNA expression of EphA2 and KLF5 in BLBC patients (n=783) (R=0.3); (K-L) Detection by tissue microarray in 86 cases revealed that EphA2 and KLF5 were both highly expressed in BLBC and that their expression was positively correlated.

Article Snippet: HCC1806 cells (1×10 6 ) were resuspended in Matrigel (#354234, Corning, BD Biocoat; 1:1 diluted with RPMI-1640 serum-free medium) and injected into the fourth pair of mammary gland fat pads.

Techniques: Expressing, Knockdown, RNA Sequencing, Over Expression, ChIP-sequencing, Binding Assay, Biomarker Discovery, Sequencing, Luciferase, Activity Assay, Microarray

Journal: International Journal of Biological Sciences

Article Title: Targeting the KLF5-EphA2 axis can restrain cancer stemness and overcome chemoresistance in basal-like breast cancer

doi: 10.7150/ijbs.82567

Figure Lengend Snippet: TNF-α induces stemness of BLBC cells partially by activating KLF5-EphA2. (A-E) Knockdown of EphA2 and KLF5 in HCC1937 and HCC1806, respectively, with concomitant addition of TNF-α for WB assay of EphA2 and KLF5 expression (A), ALDH + cell ratio by flow assay (B-C), Detection of the sphere-forming rate of HCC1937 by microsphere-formation assay (D-E), Scale bar=250 μm; (F-J) Knockdown of KLF5 and reversion to overexpression of EphA2 and addition of TNF-α in HCC1937 and HCC1806 for WB detection of EphA2 and KLF5 expression (F), ALDH + cell ratio by flow assay (G-H), Detection of the sphere-forming rate of HCC1937 by mamosphere formation assay (I-J), Scale bar=250 μm. Changed folds in (A) and (F) of indicated proteins were referenced to Control (siNC or pCDH), normalized to GAPDH, respectively.

Article Snippet: HCC1806 cells (1×10 6 ) were resuspended in Matrigel (#354234, Corning, BD Biocoat; 1:1 diluted with RPMI-1640 serum-free medium) and injected into the fourth pair of mammary gland fat pads.

Techniques: Knockdown, Expressing, Tube Formation Assay, Over Expression, Control

Journal: International Journal of Biological Sciences

Article Title: Targeting the KLF5-EphA2 axis can restrain cancer stemness and overcome chemoresistance in basal-like breast cancer

doi: 10.7150/ijbs.82567

Figure Lengend Snippet: Inhibiting EphA2 phosphorylation by ALW-II-41-27 impedes BLBC cell stemness and chemoresistance. (A-B) Detection of sphere formation by mamosphere formation assay after treating HCC1937 cells with chemotherapeutic drugs PTX (2 μM) and DDP (2 μM) alone or in combination with ALW-II-41-27 (50 nM) for 48 h. Scale bar=250 μm. (C-D) Detection of the ALDH + cell ratio by flow cytometry after treatment of HCC1937 and HCC1806 cells with the chemotherapeutic agents PTX (2 μM) or DDP (2 μM) alone or in combination with ALW-II-41-27 (50 nM) for 48 h; (E-F) Detection of EphA2 and EphA2 pS897 expression by WB after treatment of HCC1937 and HCC1806 cells with the chemotherapeutic agents PTX (2 μM) or DDP (2 μM) alone or in combination with ALW-II-41-27 (50 nM) for 48 h; (G-H) IC50 assay in HCC1937 and HCC1806 cells treated with chemotherapeutic agents PTX or DDP alone or in combination with ALW-II-41-27. Changed folds in (E) and (F) of indicated proteins were referenced to DMSO, normalized to GAPDH, respectively.

Article Snippet: HCC1806 cells (1×10 6 ) were resuspended in Matrigel (#354234, Corning, BD Biocoat; 1:1 diluted with RPMI-1640 serum-free medium) and injected into the fourth pair of mammary gland fat pads.

Techniques: Phospho-proteomics, Tube Formation Assay, Flow Cytometry, Expressing

Journal: International Journal of Biological Sciences

Article Title: Targeting the KLF5-EphA2 axis can restrain cancer stemness and overcome chemoresistance in basal-like breast cancer

doi: 10.7150/ijbs.82567

Figure Lengend Snippet: EphA2 pS897 contributes to BLBC cell chemoresistance and indicates the NCT response. (A) Detection of EphA2, KLF5 and EphA2 pS897 expression by WB after knocking down KLF5 in HCC1937 and HCC1806 cells while overexpressing EphA2 WT or EphA2 S897A and adding TNF-α(10 ng/mL) and ALW-II-41-27 (ALW; 50 nM); (B-E) Detection of the proliferation of HCC1806 and HCC1937 cells under PTX (B-C) or DDP (D-E) treatment. (F) EphA2 expression in samples from BLBC patients (BCIP database) with pCR and residual disease (RD) post neoadjuvant chemotherapy (NCT). (G) IHC detection of EphA2 pS897 expression in neoadjuvant chemoresistant (Group 1, N=11) and sensitive (Group 2, N=19) breast cancer tissue samples; (H-I) Statistics of IOD score (H) and pathological grading (I) for samples from (G). * or ns indicates the significance between the two and # indicates the significance between any one of each groups.

Article Snippet: HCC1806 cells (1×10 6 ) were resuspended in Matrigel (#354234, Corning, BD Biocoat; 1:1 diluted with RPMI-1640 serum-free medium) and injected into the fourth pair of mammary gland fat pads.

Techniques: Expressing

Journal: Technology in Cancer Research & Treatment

Article Title: CAMSAP3 Suppresses Breast Cancer Metastasis and Serves as an Independent Prognostic Marker

doi: 10.1177/15330338251381866

Figure Lengend Snippet: CAMSAP3 Overexpression Suppresses Migration and Invasion of MDA-MB-231 Breast Cancer Cells. (A) Relative CAMSAP3 mRNA Expression in MDA-MB-231 and HCC1937 Cells, as Determined by RT-qPCR. (B) WB Showing Increased CAMSAP3 Protein Expression in MDA-MB-231 Cells Following Plasmid-Mediated Overexpression. (C) Wound Healing Assay Indicating Reduced Migratory Capacity Upon CAMSAP3 overexpression. (D) Transwell invasion assay demonstrating suppressed invasion in CAMSAP3- overexpressing cells. All in vitro Experiments were Independently Repeated Three Times. Data are Presented as Mean ± SD. * P < 0.05, ** P < 0.01, *** P < 0.001. CAMSAP3, Calmodulin-Regulated Spectrin-Associated Protein 3; RT-qPCR, Real-Time Quantitative PCR; WB, Western Blot.

Article Snippet: HCC1937 cells (Procell Life Science & Technology Co., Ltd, CL-0093, RRID: CVCL_0290, Wuhan, China) were derived from a primary breast ductal carcinoma, and their identity was confirmed by the vendor via STR profiling.

Techniques: Over Expression, Migration, Expressing, Quantitative RT-PCR, Plasmid Preparation, Wound Healing Assay, Transwell Invasion Assay, In Vitro, Real-time Polymerase Chain Reaction, Western Blot

Journal: Technology in Cancer Research & Treatment

Article Title: CAMSAP3 Suppresses Breast Cancer Metastasis and Serves as an Independent Prognostic Marker

doi: 10.1177/15330338251381866

Figure Lengend Snippet: Knockdown of CAMSAP3 Enhances Migration and Invasion in HCC1937 Breast Cancer Cells. (A) WB Confirming Reduced CAMSAP3 Protein Expression Following siRNA-Mediated Knockdown. (B) Wound Healing Assay Showing Increased Migratory Capacity after CAMSAP3 Knockdown. (C) Transwell Invasion Assay Indicating Enhanced Invasion Upon CAMSAP3 Depletion. All In Vitro Experiments were Independently Repeated Three Times. Data are Presented as Mean ± SD. * P < 0.05, ** P < 0.01. CAMSAP3, Calmodulin-Regulated Spectrin-Associated Protein 3; WB, Western Blot.

Article Snippet: HCC1937 cells (Procell Life Science & Technology Co., Ltd, CL-0093, RRID: CVCL_0290, Wuhan, China) were derived from a primary breast ductal carcinoma, and their identity was confirmed by the vendor via STR profiling.

Techniques: Knockdown, Migration, Expressing, Wound Healing Assay, Transwell Invasion Assay, In Vitro, Western Blot

Journal: Antibody Therapeutics

Article Title: Affinity-optimized TROP2 antibodies support potent antitumor activity in antibody–drug conjugates

doi: 10.1093/abt/tbag011

Figure Lengend Snippet: Auristatin-based dual-payload anti-TROP2 ADCs exert remarkable antitumor effects in orthotopic xenograft mouse models of human breast cancer. (A–C) Antitumor activity (A), survival benefit (B), and body weight change (C) in HCC1806 model: A single dose of unconjugated mAb control (black circle), SG surrogate (light green diamond), MMAE/F DAR 4 + 2 dual-payload TROP2–1 ADC (cyan triangle), MMAE/F DAR 4 + 2 dual-payload Sacituzumab ADC (magenta square) or DuoDM/MMAE DAR 4 + 2 dual-payload TROP2–1 ADC (light purple diamond) was intravenously administered at 3 mg/kg to tumor-bearing female Nu/J mice at a mean tumor volume of 251.0 ± 106.4 mm 3 (n = 5 for all groups). Data are presented as mean values ± SEM. Kaplan–Meier survival curve statistics were analyzed with a logrank (Mantel–Cox) test. DuoDM, duocarmycin DM.

Article Snippet: HCC1806 cells (1 × 10 7 cells) suspended in 100 μL of PBS were orthotopically injected into the inguinal mammary fat pad of female NU/J mice (6–8 weeks old, purchased from The Jackson Laboratory, Stock number: 002019).

Techniques: Activity Assay, Control

Journal: Antibody Therapeutics

Article Title: Affinity-optimized TROP2 antibodies support potent antitumor activity in antibody–drug conjugates

doi: 10.1093/abt/tbag011

Figure Lengend Snippet: Dual-payload TROP2–1 ADCs exert remarkable antitumor effects in an immunocompetent mouse model bearing orthotopic murine breast cancer. (A, B) Characterization of human-TROP2 transduced murine cell line. Flow cytometry analysis of TROP2 expression in EMT6-WT (red), EMT6/hTROP2 (orange), and HCC1806 (positive control, cyan) cell lines (A). Cell killing potency of TROP2–1 ADCs in hTROP2 cell line (B): We tested unconjugated N297A anti-TROP2 mAb (black circle), MMAE/F DAR 4 + 2 dual-payload ADC (cyan square), SG surrogate (light green inversed triangle). All assays were performed in triplicate. Data are presented as mean values ± SEM. (C, D) Antitumor activity (C) and survival benefit (D) in hTROP2/EMT6 model: A single dose of unconjugated mAb control (black circle), SG surrogate (light green diamond), MMAE/F DAR 4 + 2 dual-payload TROP2–1 ADC (cyan triangle), MMAE/F DAR 4 + 2 dual-payload trastuzumab ADC (isotype control, blue snowflake) or DuoDM/MMAE DAR 4 + 2 dual-payload TROP2–1 ADC (light purple square) was intravenously administered at 3 mg/kg to tumor-bearing female BALB/c mice at a mean tumor volume of 175–250 mm 3 (n = 5 for all groups). Data are presented as mean values ± SEM. Kaplan–Meier survival curve statistics were analyzed with a logrank (Mantel–Cox) test. (E–H) Tumor growth curve of individual mouse. MMAE/F DAR 4 + 2 dual-payload TROP2–1 ADC group (E), DuoDM/MMAE DAR 4 + 2 dual-payload TROP2–1 ADC group (F), and SG biosimilar group (G). (H) Mice cured with the TROP2–1 ADCs (n = 2) were rechallenged with hTROP2-negative EMT6-WT cells 51 days after initial EMT6-hTROP2 tumor inoculation. Age matched naïve mice were used as control (n = 5, black square). CR, complete remission.

Article Snippet: HCC1806 cells (1 × 10 7 cells) suspended in 100 μL of PBS were orthotopically injected into the inguinal mammary fat pad of female NU/J mice (6–8 weeks old, purchased from The Jackson Laboratory, Stock number: 002019).

Techniques: Flow Cytometry, Expressing, Positive Control, Activity Assay, Control