rna  (CLS Cell Lines Service GmbH)

Journal: RSC Advances

Article Title: Photo-immunotherapeutic PDA-Ce6-R837 nanoparticles enhance the induction of immunogenic responses in melanoma cells

doi: 10.1039/d6ra01921h

Figure Lengend Snippet: PDA-Ce6-R837 NPs induce PTT and PDT effects in melanoma cells. (A) Fluorescence imaging demonstrating the cellular uptake of PDA-Ce6-R837 NPs by SK-Mel-28 and A375 cells. Cell viability of SK-Mel-28 (B) and A375 (C) cells following treatment with increasing concentrations of PDA-Ce6-R837 NPs, as assessed by the CCK-8 assay. Values represent the mean ± SD, n = 3. * p < 0.05, ** p < 0.01, *** p < 0.001, compared with Control group. CCK-8 analysis of the effects of PDA-Ce6-R837 NPs at different concentrations on the viability of SK-Mel-28 (D) and A375 (E) cells in the presence or absence of laser irradiation. Intracellular ROS production in SK-Mel-28 (F) and A375 (G) cells after PDA-Ce6-R837 NPs treatment with or without laser irradiation. (H) Quantitative analysis of ROS levels. Values represent the mean ± SD, n = 3. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: Human melanoma A375 and SK-Mel-28 cell lines were obtained from Procell (Wuhan, China).

Techniques: Fluorescence, Imaging, CCK-8 Assay, Control, Irradiation

Journal: RSC Advances

Article Title: Photo-immunotherapeutic PDA-Ce6-R837 nanoparticles enhance the induction of immunogenic responses in melanoma cells

doi: 10.1039/d6ra01921h

Figure Lengend Snippet: PDA-Ce6-R837 NPs mediated phototherapy suppresses melanoma cells migration and invasion. (A) Representative images of wound healing assays in SK-Mel-28 cells treated with PDA-Ce6-R837 NPs with or without laser irradiation at 0 h and 24 h. (B) Quantitative analysis of wound closure in SK-Mel-28 cells under the indicated treatment conditions. (C) Representative images of wound healing assays in A375 cells treated with PDA-Ce6-R837 NPs with or without laser irradiation at 0 h and 24 h. (D) Quantitative analysis of wound closure in A375 cells under the indicated treatment conditions. (E) Representative images of Transwell invasion assays showing the invasive behavior of melanoma cells under different treatments. (F) Quantification of invaded SK-Mel-28 and A375 cells following PDA-Ce6-R837 NPs mediated phototherapy. All values represent the mean ± SD, n = 3. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: Human melanoma A375 and SK-Mel-28 cell lines were obtained from Procell (Wuhan, China).

Techniques: Migration, Irradiation

Journal: RSC Advances

Article Title: Photo-immunotherapeutic PDA-Ce6-R837 nanoparticles enhance the induction of immunogenic responses in melanoma cells

doi: 10.1039/d6ra01921h

Figure Lengend Snippet: PDA-Ce6-R837 NPs mediated phototherapy induces apoptosis in melanoma cells. (A) Representative flow cytometry plots showing apoptosis in SK-Mel-28 and A375 cells following different treatments. Quantitative analysis of apoptotic rates in SK-Mel-28 (B) and A375 (C) cells under indicated conditions. All values represent the mean ± SD, n = 3. ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: Human melanoma A375 and SK-Mel-28 cell lines were obtained from Procell (Wuhan, China).

Techniques: Flow Cytometry

Journal: RSC Advances

Article Title: Photo-immunotherapeutic PDA-Ce6-R837 nanoparticles enhance the induction of immunogenic responses in melanoma cells

doi: 10.1039/d6ra01921h

Figure Lengend Snippet: PDA-Ce6-R837 NPs mediated phototherapy induces ICD in melanoma cells. Immunofluorescence staining of CRT in SK-Mel-28 (A) and A375 (B) cells under basal conditions or following treatment with PDA-Ce6-R837 NPs with or without laser irradiation. (C) Quantification of membrane-associated CRT fluorescence intensity after treatment. Measurement of extracellular ATP (D) and HMGB1 (E) release in response to PDA-Ce6-R837 NPs with or without laser irradiation. Scale bar = 50 µm. All values represent the mean ± SD, n = 3. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: Human melanoma A375 and SK-Mel-28 cell lines were obtained from Procell (Wuhan, China).

Techniques: Immunofluorescence, Staining, Irradiation, Membrane, Fluorescence

Journal: Signal Transduction and Targeted Therapy

Article Title: Bioengineered iPSC-derived human macrophages with increased angiotensin-converting enzyme (ACE) expression suppress solid tumor growth

doi: 10.1038/s41392-026-02650-3

Figure Lengend Snippet: Determination of M1 vs. M2 polarization in iMac. a Pro-inflammatory M1 cytokines IL-1β, TNFα, IL-6, and IL-8 measured by ELISA. b Anti-inflammatory M2 cytokines IL-4, IL-10, IL-13, CCL5, CCL17, and CCL20 measured by ELISA. For a , b iMac were cultured in 6-well plates (3 × 10 6 cells/well). To challenge with tumor factors, iMac were co-cultured with 50% fresh supernatant from SK-MEL-28 cells every 24 h for 3 days. Supernatants were collected during the final 24 h for cytokine measurement. c Activation (phosphorylation) of NF-kB p65, STAT1, STAT3, and STAT6 in iMac was determined using Western blotting after conditioning with melanoma (SK-MEL-28) supernatant for 72 h. Protein band intensity was quantified using Odyssey version 3.0 software. After adjusting for loading using the β-actin band, data are presented as fold change to the iMac (Dox-) group, which is set as 1. For a – c , one-way ANOVA with Bonferroni’s correction for multiple comparisons was used to analyze group comparisons. Data are presented as means ± SD. Each experiment was performed three times, with triplicate samples each time. * p < 0.05, *** p < 0.001, and **** p < 0.0001

Article Snippet: The triple negative breast (TNB) cancer cell line HCC1806 and the melanoma cell line SK-MEL-28 were obtained from ATCC.

Techniques: Enzyme-linked Immunosorbent Assay, Cell Culture, Activation Assay, Phospho-proteomics, Western Blot, Software

Journal: Signal Transduction and Targeted Therapy

Article Title: Bioengineered iPSC-derived human macrophages with increased angiotensin-converting enzyme (ACE) expression suppress solid tumor growth

doi: 10.1038/s41392-026-02650-3

Figure Lengend Snippet: Determination of anti-tumor activity of iMac with induced ACE expression, in vitro. a Proliferation of melanoma (SK-MEL-28), Triple Negative Breast Cancer (TNBC: HCC1806), and FP chemotherapy resistant HNSCC (FaDu/FP-R) cell lines cultured alone or indirectly co-cultured with iMac ( ± Dox) using 3.0 µm pore polyester membrane inserts for 5 days. Total and dead cell counts were determined via trypan blue exclusion on days 1, 3, and 5. b , c Quantification of iNOS and interleukin-12 (IL-12) production in Cnt-iMac and ACE-iMac treated with LPS (500 ng/ml, 12 h), ± Dox. d ROS production in ACE-iMac following LPS stimulation (1 µg/ml, 30 min) ±Dox, assessed by DCFDA (2’,7’-dichlorodihydrofluorescein diacetate) staining. Representative histograms (left) and mean fluorescence intensity (MFI) quantification (right) are shown. Nitric Oxide (NO) production in ACE-iMac stimulated with LPS (75 ng/ml) for 24 h ( e ), and/or conditioned with SK-MEL-28 supernatant for 24 h ( f ) ± Dox. g Perforin and IFN-γ levels in human peripheral blood NK and Tc cells, respectively, after co-culture with Cnt-iMac or ACE-iMac pretreated with LPS (24 h), followed by melanoma conditioning (24 h). To induce ACE expression, myeloid progenitors were differentiated in the presence of Dox and maintained at 1 µg/ml throughout all in vitro assays. Experiments were performed in triplicates across three independent replicates. Statistical analyses included one-way ANOVA ( a ), two-way ANOVA with Bonferroni correction ( b , c , g ), and two-sided unpaired Student’s t -test ( d – f ). Data are presented as means ± SD. * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001

Article Snippet: The triple negative breast (TNB) cancer cell line HCC1806 and the melanoma cell line SK-MEL-28 were obtained from ATCC.

Techniques: Activity Assay, Expressing, In Vitro, Cell Culture, Membrane, Staining, Fluorescence, Co-Culture Assay

Journal: Signal Transduction and Targeted Therapy

Article Title: Bioengineered iPSC-derived human macrophages with increased angiotensin-converting enzyme (ACE) expression suppress solid tumor growth

doi: 10.1038/s41392-026-02650-3

Figure Lengend Snippet: The effect of ACE-expressing iMac treatment on tumor growth in immunocompromised nude mice. To induce ACE expression, myeloid progenitors were differentiated in the presence of 1 µg/ml Dox (termed as ACE-iMac), while control iMac with basal ACE expression were differentiated in the absence of Dox. a Experimental scheme of iMac treatment. Mice were injected intratumorally with PBS, Cnt-iMac (±Dox) or ACE-iMac (±Dox) weekly (3–4 doses) as indicated by arrow. b Growth of melanoma xenografts derived from the SK-MEL-28 cell line. c Growth of breast cancer xenografts derived from the HCC1806 cell line. d Growth of HNSCC xenografts derived from the FaDu/FP-R cell line. For b – d Left panels show plots comparing tumor growth rates, the middle panels show representative images of tumors, and right panels show graphs comparing tumor weight. Following cell administration, Dox+ (ACE-iMac) mouse groups were provided with 150 µg/ml doxycycline in their drinking water throughout the duration of the study. Group comparisons were analyzed using one-way ANOVA with Bonferroni’s correction for multiple comparisons. Data are presented as means ± SEM ( n = 5–10). * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001

Article Snippet: The triple negative breast (TNB) cancer cell line HCC1806 and the melanoma cell line SK-MEL-28 were obtained from ATCC.

Techniques: Expressing, Control, Injection, Derivative Assay

Journal: Signal Transduction and Targeted Therapy

Article Title: Bioengineered iPSC-derived human macrophages with increased angiotensin-converting enzyme (ACE) expression suppress solid tumor growth

doi: 10.1038/s41392-026-02650-3

Figure Lengend Snippet: Analysis of anti-tumor macrophage activation in the TME post-iMac treatment. Tumors were harvested and single-cell suspensions were prepared for flow cytometry analysis as detailed in the methods section (refer to Fig. ). a Gating strategy for distinguishing M1 and M2 macrophages using flow cytometry based on surface marker immunostaining. b Assessment of CD86 + (M1) and CD206 + (M2) expression in tumor associated macrophages (TAMs) in SK-MEL-28-derived xenografts (±Dox). The bar plot presents FACS quantification of M1 and M2 populations ( n = 5/group). c Assessment of arginase expression in TAMs. d ACE expression in TAMs. e – g Measurement of proinflammatory and anti-tumor markers IFN-γ, IL-12, and iNOS (left: representative histogram; right: mean fluorescence intensity) in TAMs in melanoma xenografts ( n = 5/group). Statistical analysis was performed using a two-sided unpaired Student’s t -test for comparisons in ( b , d , g ), and one-way ANOVA with Bonferroni’s correction for multiple comparisons in ( c , e , f ). Data are presented as means ± SEM. * p < 0.05 and **** p < 0.0001

Article Snippet: The triple negative breast (TNB) cancer cell line HCC1806 and the melanoma cell line SK-MEL-28 were obtained from ATCC.

Techniques: Activation Assay, Single Cell, Flow Cytometry, Marker, Immunostaining, Expressing, Derivative Assay, Fluorescence

Journal: Signal Transduction and Targeted Therapy

Article Title: Bioengineered iPSC-derived human macrophages with increased angiotensin-converting enzyme (ACE) expression suppress solid tumor growth

doi: 10.1038/s41392-026-02650-3

Figure Lengend Snippet: Analysis of NK cell and B cell activation in the TME post-iMac treatment. Tumors were harvested, and single-cell suspensions were prepared for flow cytometry analysis. a Representative gating strategy for identifying activated NK cells, B cells, and neutrophils in tumor xenografts using flow cytometry. b Percentage of activated NK cells (CD122 + CD107a + ) in melanoma, breast cancer, and HNSCC xenografts ( n = 5/group). c Measurement of IFN-γ-positive NK cells in Cnt-iMac and ACE-iMac melanoma tumors (±Dox) ( n = 5/group) in SK-MEL-28-derived xenografts. d Percentage of activated B cells (CD19 + CD21/CD35 + ) in melanoma, breast cancer, and HNSCC xenografts ( n = 5/group). Statistical analysis was performed using a one-way ANOVA with Bonferroni’s correction for multiple comparisons in ( b – d ). Data are presented as means ± SEM. * p < 0.05 and **** p < 0.0001

Article Snippet: The triple negative breast (TNB) cancer cell line HCC1806 and the melanoma cell line SK-MEL-28 were obtained from ATCC.

Techniques: Activation Assay, Single Cell, Flow Cytometry, Derivative Assay

Journal: Signal Transduction and Targeted Therapy

Article Title: Bioengineered iPSC-derived human macrophages with increased angiotensin-converting enzyme (ACE) expression suppress solid tumor growth

doi: 10.1038/s41392-026-02650-3

Figure Lengend Snippet: The effect of ACE-iMac treatment on human immune cells in a humanized mouse tumor model. a Growth of SK-MEL-28 tumors in humanized BLT-NSG mice; comparing tumor volumes between PBS, iMac and ACE-iMac treated groups. b Representative images of tumors are shown (see Supplementary Fig. for additional images). c Final tumor volumes on Day 28 post-implantation. d Tumor weight on Day 28 post-implantation. Flow cytometry analysis of anti-tumor activation of human CD8 T cells in the TME, measured by intracellular IFN-γ ( e ) and perforin ( f ) in tumor-infiltrating CD3 + CD8 + T cells at sacrifice on Day 28 post-implantation. g Flow cytometry analysis of anti-tumor activation of NK cells, measured by intracellular IFN-γ in tumor-infiltrating CD45 + CD56 + NK cells in the TME on Day 28. The flow cytometry gating strategy for identifying human immune cells is provided in Supplementary Fig. . One-way ANOVA with Bonferroni’s correction for multiple comparisons was used to analyze group comparisons. Data are presented as means ± SEM ( n = 9–10). * p < 0.05, ** p < 0.01, and **** p < 0.0001

Article Snippet: The triple negative breast (TNB) cancer cell line HCC1806 and the melanoma cell line SK-MEL-28 were obtained from ATCC.

Techniques: Flow Cytometry, Activation Assay

Journal: Signal Transduction and Targeted Therapy

Article Title: Bioengineered iPSC-derived human macrophages with increased angiotensin-converting enzyme (ACE) expression suppress solid tumor growth

doi: 10.1038/s41392-026-02650-3

Figure Lengend Snippet: Transcriptomic profiling and pathway analysis of ACE-iMac following tumor-stimulation. ACE-iPSCs were differentiated into iMac ± 1 µg/ml Dox. iMac were then stimulated with SK-MEL-28 conditioned medium for 12 h prior to RNA extraction and sequencing. a Volcano plot showing significantly altered mRNA expression in ACE-iMac versus iMac. b IPA of differentially expressed genes. c Bubble charts illustrating significant canonical pathways. d Heatmap of gene expression highlighting the macrophage classical activation signaling pathway. e Expression levels of key RNAs within the macrophage classical activation signaling gene cluster. f GO analysis of differentially expressed genes, with upregulated (red bars) and downregulated (blue bars) clusters ranked by pathway enrichment. g Heatmap showing differential expression of genes involved in primary metabolic processes. h Expression levels of representative RNAs from metabolic gene clusters. Statistical analyses were performed using a two-sided unpaired Student’s t -test in ( a , e , h ). Data are presented as means ± SEM. **** p < 0.0001. For details, refer to Fig.

Article Snippet: The triple negative breast (TNB) cancer cell line HCC1806 and the melanoma cell line SK-MEL-28 were obtained from ATCC.

Techniques: RNA Extraction, Sequencing, Expressing, Gene Expression, Activation Assay, Quantitative Proteomics