Journal: Scientific Reports

Article Title: The novel amino-artemisinin derivative WHN-11 disrupts mitochondria and protein homeostasis, and induces autophagy and apoptosis in cancer cells

doi: 10.1038/s41598-025-05284-7

Figure Lengend Snippet: Novel artemisinin derivatives exhibit anti-cancer and anti-cancer stem cell activity ( a ) Heatmap showing the IC 50 values (in µM) of artemisinins screened against non-cancerous MCF-12A epithelial breast cell line, TNBC cell lines: HCC1937, HCC70, MDA-MB-231; non-breast cancer cell lines: 501 melanoma, HCT116, HeLa and U87 cell lines. DHA – dihydroartemisinin ( b ) Percentage tumoursphere forming efficiency (%TFE) of cells cultured in AIG media and treated at a single-point concentration of each compound (50 µM) or DMSO for 7 days; % TFE = [total number of tumoursphere formed / number of cells seeded] × 100%. Data shown as averages ± SEM (n = 3) and statistical significance were determined using one-way ANOVA with Bonferroni’s multiple comparisons test. Significance levels relative to DMSO-treatment are indicated as * p < 0.05, ** p < 0.01. ( c ) Representative images of colonies formed in a clonogenic assay for treated HCC1937 cells cultured over 9 days. Following solubilization, average absorbance at 570 nm (A 570nm ) ± SEM (n = 3) was measured. Statistical significance was assessed by one-way ANOVA with Tukey’s multiple comparison test, where significance levels are indicated as *** p < 0.001 and ns—not significant. 5FU: 5-fluorouracil ( d ) Caspase activation in treated HCC1937 cells using Vybrant™ FAM Poly Caspase Assay Kit. Data shown are MFI average ± SEM (n = 3), where significance levels are indicated as * p < 0.05, *** p < 0.001 and ns—not significant using a two-tailed student’s t-test analysis. ( e ) Western blot analysis of PARP-1 and Caspase-3 activation in treated cells, tubulin was used as a loading control. GA: geldanamycin (see Supplementary blot figure for full length blots).

Article Snippet: The triple negative breast cancer cell lines HCC1937 (ATCC: CRL-2336), HCC70 (ATCC: CRL-2315), MDA-MB-231 (ATCC: HTB-26), the cervical carcinoma cell line HeLa (ATCC: CCL-2), and colon cancer cell line HCT116 (ATCC: CCL-247) were purchased from the ATCC.

Techniques: Activity Assay, Cell Culture, Concentration Assay, Clonogenic Assay, Comparison, Activation Assay, Caspase Assay, Two Tailed Test, Western Blot, Control

Journal: Scientific Reports

Article Title: The novel amino-artemisinin derivative WHN-11 disrupts mitochondria and protein homeostasis, and induces autophagy and apoptosis in cancer cells

doi: 10.1038/s41598-025-05284-7

Figure Lengend Snippet: WHN-11 promotes protein ubiquitination and turnover in HCC1937 cells. ( a ) Confocal analysis staining of ubiquitin in untreated cells or cells treated with 2 µM MG132. Representative images shown (n = 3). ( b ) Mean of the Corrected Total Cell Fluorescence (CTFC). Data represent averages ± SD (n = 3), where significance levels are indicated as ** p < 0.01, *** p < 0.001 and ns–not significant using two-way ANOVA with Bonferroni post-test. ( c ) Direct chymotrypsin-like proteasome inhibition activity assay in cell lysates. ( d ) Proteasome activity after overnight pre-treatment of live cells prior to lysate preparation. Data shown are averages ± SEM (n = 3), where significance levels are indicated as *** p < 0.001 and ns—not significant using One-way ANOVA with Bonferroni’s multiple comparison test. ( e ) Quantitative analysis of the mRNA copy number of Hsp70 in treated HCC1937 cells. Data shown are averages ± SEM (n = 3), where significance levels are defined as * p < 0.05, ** p < 0.01 and ns—not significant using a two-tailed student´s t-test.

Article Snippet: The triple negative breast cancer cell lines HCC1937 (ATCC: CRL-2336), HCC70 (ATCC: CRL-2315), MDA-MB-231 (ATCC: HTB-26), the cervical carcinoma cell line HeLa (ATCC: CCL-2), and colon cancer cell line HCT116 (ATCC: CCL-247) were purchased from the ATCC.

Techniques: Ubiquitin Proteomics, Staining, Fluorescence, Inhibition, Activity Assay, Comparison, Two Tailed Test

Journal: Scientific Reports

Article Title: The novel amino-artemisinin derivative WHN-11 disrupts mitochondria and protein homeostasis, and induces autophagy and apoptosis in cancer cells

doi: 10.1038/s41598-025-05284-7

Figure Lengend Snippet: WHN-11 promotes formation of autophagic vesicles and lipid droplets in HCC1937 cells. ( a ) Acidic vesicular organelles (AVOs, white arrows) detected with acridine orange dye in cells treated for 4 h with 0.1% DMSO, 10 µM WHN-11 or Hanks’ Balanced Salt Solution (HBSS). Yellow arrows indicate unstained vesicles. Quantitation of ( b ) fluorescence intensity of AVOs and ( c ) number of cells with AVOs. Data represent averages ± SEM (n = 3), where significance levels are indicated as * p < 0.05, ** p < 0.01 and *** p < 0.001 using a One-way ANOVA. ( d ) Lipid droplets (white arrows) stained with Nile Red dye in cells treated for 4 h with 0.1% DMSO, HBSS or 10 µM WHN-11. Z depth panel shows 3D z-stack of equivalent depth. Representative images shown (n = 3).

Article Snippet: The triple negative breast cancer cell lines HCC1937 (ATCC: CRL-2336), HCC70 (ATCC: CRL-2315), MDA-MB-231 (ATCC: HTB-26), the cervical carcinoma cell line HeLa (ATCC: CCL-2), and colon cancer cell line HCT116 (ATCC: CCL-247) were purchased from the ATCC.

Techniques: Quantitation Assay, Fluorescence, Staining

Journal: Scientific Reports

Article Title: The novel amino-artemisinin derivative WHN-11 disrupts mitochondria and protein homeostasis, and induces autophagy and apoptosis in cancer cells

doi: 10.1038/s41598-025-05284-7

Figure Lengend Snippet: Activation of apoptotic and autophagic signalling pathways in WHN-11-treated HCC1937 cells. ( a ) Western blot analysis of LC3B lipidation in cells treated overnight with 10 µM WHN-11 and for 4 h with 2 µM chloroquine (CQ). Actin was used as a loading control. Ratios indicated are normalized to the untreated control. Representative blot shown (n = 2). See supplementary blot figure for full length blots. ( b ) Time-dependent treatment of cells with 0.1% DMSO or WHN-11 for caspase-3 and LC3B activation. Actin was used as a loading control. Ratios of LC3B-II: LC3B-I and cCas-3:Cas-3 are normalized to the untreated control. Representative blot shown (n = 2). See supplementary blot figure for full length blots. ( c ) Immunoprecipitation of GFP or GFP-Bcl2 complexes in lysates (input) for HEK293T cells transfected with pLV-eGFP (untreated control) or GFP-Bcl2 and Beclin1-FLAG for 48 h and treated for 3 h with 0.1% DMSO, Hanks’ Balanced Salt Solution (HBSS), 5 µM colchicine or 10 µM WHN-11. Representative blots shown. Analysis of the average fold difference ± SEM (n = 3) of Beclin1-FLAG levels in immunoprecipitates relative to GFP-Bcl2 and normalized to DMSO treatment (taken as 1), where significance levels are indicated as * p < 0.05 and ** p < 0.01 using a two-tailed student’s t-test. See supplementary blot figure for full length blots.

Article Snippet: The triple negative breast cancer cell lines HCC1937 (ATCC: CRL-2336), HCC70 (ATCC: CRL-2315), MDA-MB-231 (ATCC: HTB-26), the cervical carcinoma cell line HeLa (ATCC: CCL-2), and colon cancer cell line HCT116 (ATCC: CCL-247) were purchased from the ATCC.

Techniques: Activation Assay, Western Blot, Control, Immunoprecipitation, Transfection, Two Tailed Test

Journal: Scientific Reports

Article Title: The novel amino-artemisinin derivative WHN-11 disrupts mitochondria and protein homeostasis, and induces autophagy and apoptosis in cancer cells

doi: 10.1038/s41598-025-05284-7

Figure Lengend Snippet: WHN-11 disrupts mitochondrial function. ( a ) Analysis of Bcl-xL expression in treated HCC1937 cells overnight. Data indicate the mean fluorescence intensity ± SEM (n = 3), where significance levels are indicated as * p < 0.05 using a two-tailed student’s t-test. ( b ) Cellular ATP levels quantified in treated cells. Data shown are averages ± SEM (n = 3), where significance levels are indicated as * p < 0.05, ** p < 0.01 and *** p < 0.001 using a two-tailed student’s t-test. ( c ) Mitochondrial membrane potential (ΔΨm) measured by JC-1 staining in treated HCC1937 cells. Data shown are averages ± SEM (n = 3) of the relative percentage ratio of red to green fluorescence, where significance levels are indicated as ** p < 0.01, *** p < 0.001 and ns–not significant using a two-tailed student’s t-test. ( d ) Confocal analysis of the mitochondrial morphology indicated by TRAP1 and Bcl-xL staining in treated HCC1937 cells. Representative images shown (n = 2).

Article Snippet: The triple negative breast cancer cell lines HCC1937 (ATCC: CRL-2336), HCC70 (ATCC: CRL-2315), MDA-MB-231 (ATCC: HTB-26), the cervical carcinoma cell line HeLa (ATCC: CCL-2), and colon cancer cell line HCT116 (ATCC: CCL-247) were purchased from the ATCC.

Techniques: Expressing, Fluorescence, Two Tailed Test, Membrane, Staining

Journal: Veterinary Sciences

Article Title: Tumor Growth Progression in Ectopic and Orthotopic Xenografts from Inflammatory Breast Cancer Cell Lines

doi: 10.3390/vetsci8090194

Figure Lengend Snippet: Tumor growth parameters of IPC-366 and SUM149 cell lines in ectopic and orthotopic models.

Article Snippet: The human triple-negative inflammatory breast cancer cell line SUM149 was obtained from Asterand, Inc. (Detroit, MI, USA), (RRID: CVCL_3422).

Techniques: Injection

Journal: Veterinary Sciences

Article Title: Tumor Growth Progression in Ectopic and Orthotopic Xenografts from Inflammatory Breast Cancer Cell Lines

doi: 10.3390/vetsci8090194

Figure Lengend Snippet: IPC-366 and SUM149 xenotransplanted mice, paraffin sections, H-E. ( A ) IPC-366 ectopic xenotransplanted mice. Neoplastic cells arranged in solid masses separated by a scant fibrovascular stroma infiltrating the adjacent dermis (inset: neoplastic cells infiltrating adjacent dermis). ( B ) IPC-366 orthotopic mice. Unencapsulated and densely cellular mass extending into the adjacent adipose tissue. ( C , D ) Ectopic and orthotopic IPC-366 xenotransplanted mice. Tumors are composed of highly pleomorphic cells with marked anisocytosis and anisokaryosis. Binucleated cells are commonly seen (arrow). ( E , F ) Ectopic and orthotopic SUM149 xenografted mice. Solid tumors infiltrate the dermis and adipose tissue. No histological differences were found between the types of SUM149 xenografts. ( G ) Orthotopic SUM149 xenograft. Medium to large round cells with a moderate eosinophilic cytoplasm and large nuclei with one or more evident nucleoli. ( H ) Orthotopic SUM149 xenograft. Presence of neoplastic cells with an elongated and empty cytoplasm that displaced the nuclei to the periphery, suggestive of endothelial-like cells (ELCs) (arrow). Atypical mitoses were frequently seen (arrowhead).

Article Snippet: The human triple-negative inflammatory breast cancer cell line SUM149 was obtained from Asterand, Inc. (Detroit, MI, USA), (RRID: CVCL_3422).

Techniques:

Journal: Veterinary Sciences

Article Title: Tumor Growth Progression in Ectopic and Orthotopic Xenografts from Inflammatory Breast Cancer Cell Lines

doi: 10.3390/vetsci8090194

Figure Lengend Snippet: Estrogen receptor (ER), Progesterone receptor (PR), and human epidermal growth factor receptor 2 (HER-2) expression on ectopic and orthotopic xenografts from IPC-366 and SUM149 cell lines.

Article Snippet: The human triple-negative inflammatory breast cancer cell line SUM149 was obtained from Asterand, Inc. (Detroit, MI, USA), (RRID: CVCL_3422).

Techniques: Expressing

Journal: Veterinary Sciences

Article Title: Tumor Growth Progression in Ectopic and Orthotopic Xenografts from Inflammatory Breast Cancer Cell Lines

doi: 10.3390/vetsci8090194

Figure Lengend Snippet: Steroid hormone secretion studied (P4, DHEA, A4, T, DHT, E1SO4, and E2), on ectopic (subcutaneous) and orthotopic (mammary fat pad) models of IPC-366 and SUM149 xenografts.

Article Snippet: The human triple-negative inflammatory breast cancer cell line SUM149 was obtained from Asterand, Inc. (Detroit, MI, USA), (RRID: CVCL_3422).

Techniques:

Journal: Frontiers in Pharmacology

Article Title: Multifaced roles of cannabinoid therapy in cancer: balancing analgesia, antitumor potential, and systemic toxicity

doi: 10.3389/fphar.2025.1691893

Figure Lengend Snippet: Experimental design. In vitro: 4T1 breast cancer cells were treated with PTX, JWH-182, NC-1, and NC-2, alone or in combination and cell viability was assessed via MTT assay. In vivo: BALB/c mice were randomized into four treatment groups (PTX alone or combined with cannabinoids), and pain was assessed using the Hot Plate, Tail Flick, and Randall-Selitto tests on days 1, 8, and 26. Tumor progression was monitored at baseline and end of experiment via breast ultrasound and MRI scans.

Article Snippet: The 4T1 cell line (triple negative breast cancer) was obtained from ATCC (Virginia, United States).

Techniques: In Vitro, MTT Assay, In Vivo, Tail Flick Test, Randall–Selitto Test

Journal: Frontiers in Pharmacology

Article Title: Multifaced roles of cannabinoid therapy in cancer: balancing analgesia, antitumor potential, and systemic toxicity

doi: 10.3389/fphar.2025.1691893

Figure Lengend Snippet: Dose-response curves indicate that PTX exhibits cytotoxic activity with an IC50 of 5.6 µM, while JWH-182 and NC-2 show moderate antitumor effects, with IC50 values of 23.2 µM and 21.1 µM, respectively. NC-1 displays minimal antitumor activity, achieving only 11% cell viability reduction at its highest dose. Combined treatments reveal that cannabinoids enhance PTX’s cytotoxic efficacy, with the PTX & JWH-182 combination showing the greatest synergistic reduction in tumor cell viability. The experiment was performed in triplicate using the 4T1 breast cancer cell line.

Article Snippet: The 4T1 cell line (triple negative breast cancer) was obtained from ATCC (Virginia, United States).

Techniques: Activity Assay

Journal: Frontiers in Pharmacology

Article Title: Multifaced roles of cannabinoid therapy in cancer: balancing analgesia, antitumor potential, and systemic toxicity

doi: 10.3389/fphar.2025.1691893

Figure Lengend Snippet: Results of complete blood count (white blood cells, neutrophils, lymphocites, monocytes, eosinophils, basophils, red blood cells, hemoglobyn, hematocrit, platelets) performed at the end of study treatments. Increased white blood cells among all groups caused by leukemoid reaction, produced by the 4T1 cell line. High levels of lymphocites among NC1 and NC2 treated animals.

Article Snippet: The 4T1 cell line (triple negative breast cancer) was obtained from ATCC (Virginia, United States).

Techniques: Produced