Journal: Cancer Cell International

Article Title: Dual-biomarkers encoding tumor-activatable minicircles with scaffold/matrix attachment region motif for sensitive and sustained blood and urine-based cancer detection

doi: 10.1186/s12935-026-04241-2

Figure Lengend Snippet: hCG and Gluc are reliable biomarkers with minimal background signals. HeLa cells were either untransfected or transfected with EF1 promoter-driven plasmids (PPs) encoding hCG together with eGFP (PP-pEF1-eGFP-M18) or Gluc (PP-pEF1-Gluc-M18). ( A ) Secreted hCG levels in culture supernatants were quantified by ELISA. Both transfected groups exhibited significantly higher hCG levels compared with untransfected controls. ( B ) Gluc level in supernatants was measured by a luminometer. Robust signals were detected only in the Gluc-transfected group. ( C ) Intracellular Gluc expression was visualized by bioluminescence imaging (BLI), again restricted to Gluc-transfected cells. Together, these results demonstrate that Gluc functions as a sensitive biomarker in both liquid assays and imaging applications. ᵃP < 0.001 vs. untransfected control; ᵇP < 0.001 vs. PP-pEF1-M18

Article Snippet: The human chorionic gonadotropin beta DuoSet ELISA kit (Cat# DY9034; R&D Systems) was used to quantify hCG levels in cell culture supernatants, urine, and plasma, in accordance with the manufacturer’s instructions.

Techniques: Transfection, Enzyme-linked Immunosorbent Assay, Expressing, Imaging, Biomarker Discovery, Control

Journal: Cancer Cell International

Article Title: Dual-biomarkers encoding tumor-activatable minicircles with scaffold/matrix attachment region motif for sensitive and sustained blood and urine-based cancer detection

doi: 10.1186/s12935-026-04241-2

Figure Lengend Snippet: Transfection with MC-pSURV-M18 restores biomarker expression reduced by promoter swapping. HeLa cells were transfected with PP-pEF1-M18, PP-pSURV-M18, or MC-pSURV-M18 to evaluate the effects of promoter swapping and minicircle constructs on biomarker expression. ( A ) hCG ELISA showed that replacing the constitutive EF1 promoter with the tumor-specific SURVIVIN promoter substantially reduced hCG expression. However, MC-pSURV-M18 transfection restored hCG expression to levels comparable to PP-pEF1-M18. ( B ) Gluc assay and ( C ) Gluc imaging demonstrated similar trends: promoter swapping decreased expression, whereas the corresponding minicircle construct rescued the loss. ᵃP < 0.001 vs. untransfected control; ᵇP < 0.001 vs. PP-pEF1-M18; ᶜP < 0.001 vs. PP-pSURV-M18

Article Snippet: The human chorionic gonadotropin beta DuoSet ELISA kit (Cat# DY9034; R&D Systems) was used to quantify hCG levels in cell culture supernatants, urine, and plasma, in accordance with the manufacturer’s instructions.

Techniques: Transfection, Biomarker Discovery, Expressing, Construct, Enzyme-linked Immunosorbent Assay, Imaging, Control

Journal: Cancer Cell International

Article Title: Dual-biomarkers encoding tumor-activatable minicircles with scaffold/matrix attachment region motif for sensitive and sustained blood and urine-based cancer detection

doi: 10.1186/s12935-026-04241-2

Figure Lengend Snippet: Tumor-activatable MCs encoding hCG and Gluc distinguish tumor-bearing mice from healthy controls. HeLa/Luc2-eGFP tumor-bearing mice were transfected in vivo with MC-pSURV-hCG-P2A-Gluc (MC) or MC-pSURV-hCG-P2A-Gluc-M18 (MC-M18). ( A ) Longitudinal Gluc imaging showed that MC-M18 sustained biomarker expression more effectively than MC. The hCG levels in ( B ) urine and ( C ) blood, measured by ELISA, were significantly elevated after transfection with either tumor-activatable MC, with or without the M18 S/MAR motif. ( D ) Urine and ( E ) blood Gluc levels showed greater variability and fluctuations; however, significant differences were detected between tumor-bearing mice transfected with MC (or MC-M18) and their respective healthy controls at specific time points. * p < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001. * indicates comparing with the Control + MC-M18 group; # indicates comparing with the Control + MC group

Article Snippet: The human chorionic gonadotropin beta DuoSet ELISA kit (Cat# DY9034; R&D Systems) was used to quantify hCG levels in cell culture supernatants, urine, and plasma, in accordance with the manufacturer’s instructions.

Techniques: Transfection, In Vivo, Imaging, Biomarker Discovery, Expressing, Enzyme-linked Immunosorbent Assay, Control

Journal: Frontiers in Immunology

Article Title: Decidualization of Stromal Cells Promotes Involvement of Mast Cells in Successful Human Pregnancy by Increasing Stem Cell Factor Expression

doi: 10.3389/fimmu.2022.779574

Figure Lengend Snippet: The migration and LIF production of the human mast cell line LAD2 are increased when co-cultured with decidualized T HESCs compared with non-decidualized T HESCs. (A) Two-chamber assay (n = 3). The upper chambers containing LAD2 cells were inserted into the lower chambers, which contained culture medium only (“none”), or decidualized or non-decidualized T HESCs cells. After an 8-hour incubation, the cells migrating into the lower chambers were counted. * P < 0.05 compared with none and ** P < 0.05 compared with non-decidualized T HESCs in the lower chamber. (B) LIF ELISA (n = 5). LAD2 cells were cultured in cytokine-free medium only (“none”), medium with SCF (“none + SCF”), and cytokine-free medium with non-decidualized or decidualized T HESCs for 8 (h) The supernatants were collected and used in ELISA kits. * P < 0.05 compared with none and ** P < 0.05 compared with none + SCF.

Article Snippet: In addition, aliquots (100 μL) of the supernatants were also collected from 5 × 10 4 HTR-8/SVneo cells cultured with 20 μM forskolin (FK; Calbiochem, San Diego, CA, USA) in 200 μL of cytokine-free StemPro-34 for 48 h. The aliquots were used in ELISA kits (Free beta-CG (Human) ELISA Kit; #KA0210; Abnova, Taipei, Taiwan) according to the manufacturer’s protocol.

Techniques: Migration, Cell Culture, Boyden Chamber Assay, Incubation, Enzyme-linked Immunosorbent Assay

Journal: Frontiers in Immunology

Article Title: Decidualization of Stromal Cells Promotes Involvement of Mast Cells in Successful Human Pregnancy by Increasing Stem Cell Factor Expression

doi: 10.3389/fimmu.2022.779574

Figure Lengend Snippet: LIF derived from LAD2 increases migration and extravillous trophoblast (EVT) differentiation (based on CD9 expression) and decreases syncytiotrophoblast (ST) differentiation (based on β human chorionic gonadotropin [hCG] expression) in the human trophoblast cell line HTR-8/SVneo when co-cultured with decidualized T HESCs. (A) Two-chamber assay (n = 5). The upper chambers containing HTR-8/SVneo cells with control IgG or anti-LIF neutralizing antibodies were inserted into the lower chambers containing culture medium only (“none”), decidualized T HESCs or non-decidualized T HESCs cells with or without LAD2. After an 18-hour incubation, the cells that migrated to the bottom of the upper wells were counted. * P < 0.05 compared with none. (B) CD9 mRNA (n = 4). HTR-8/SVneo cells with culture supernatants of decidualized T HESCs (alone or pre-co-cultured with LAD2 cells with control IgG or anti-LIF antibody for 48 h) were collected, and mRNA was extracted and utilized for real time PCR. * P < 0.05 compared with decidualized T HESCs. (C) β hCG ELISA (n = 4). The culture supernatants from decidualized T HESCs incubated with cytokine-free medium only, or with LAD2 and control IgG or anti-LIF antibody were added to HTR-8/SVneo cells. After a 24-h incubation, the supernatants were collected (“Decidualized”, “Decidualized + LAD2 + control IgG”, or “Decidualized + LAD2 + anti-LIF antibody”). In addition, the supernatants from HTR-8/SVneo cells cultured with forskolin (FK) for 48 h (“Decidualized + FK”) were also collected. * P < 0.05 compared with Decidualized.

Article Snippet: In addition, aliquots (100 μL) of the supernatants were also collected from 5 × 10 4 HTR-8/SVneo cells cultured with 20 μM forskolin (FK; Calbiochem, San Diego, CA, USA) in 200 μL of cytokine-free StemPro-34 for 48 h. The aliquots were used in ELISA kits (Free beta-CG (Human) ELISA Kit; #KA0210; Abnova, Taipei, Taiwan) according to the manufacturer’s protocol.

Techniques: Derivative Assay, Migration, Expressing, Cell Culture, Boyden Chamber Assay, Control, Incubation, Real-time Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay