skov3 cl-0215  (Procell Inc)

Journal: Biology Direct

Article Title: ALKBH5 activates CEP55 transcription through m6A demethylation in FOXP2 mRNA and expedites cell cycle entry and EMT in ovarian cancer

doi: 10.1186/s13062-024-00551-5

Figure Lengend Snippet: CEP55 inhibits the malignant behavior of OC cells. A , CEP55 expression in A2780 and SKOV3 cells infected with lentiviral vectors harboring CEP55 knockdown assessed using RT-qPCR. B , The number of A2780 and SKOV3 cells-formed colonies was assessed using colony formation assay. C , The proliferation of A2780 and SKOV3 cells was assessed using an EdU assay. D , The expression of proliferation-related genes KI67 and PCNA in A2780 and SKOV3 cells with knockdown of CEP55 was assessed using RT-qPCR. E , Percentage of apoptosis in A2780 and SKOV3 cells with knockdown of CEP55 detected by TUNEL assay. Statistical significance was assessed using ANOVA. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001. All cell experiments were repeated three times

Article Snippet: Human OC cell lines A2780 (CL-0013) and SKOV3 (CL-0215) were procured from Procell (Wuhan, Hubei, China) and cultured in RPMI-1640 medium containing 10% FBS and 1% penicillin/streptomycin in an incubator containing 5% CO 2 at 37 °C.

Techniques: Expressing, Infection, Knockdown, Quantitative RT-PCR, Colony Assay, EdU Assay, TUNEL Assay

Journal: Biology Direct

Article Title: ALKBH5 activates CEP55 transcription through m6A demethylation in FOXP2 mRNA and expedites cell cycle entry and EMT in ovarian cancer

doi: 10.1186/s13062-024-00551-5

Figure Lengend Snippet: Knockdown of CEP55 induces cell cycle arrest and suppresses EMT in OC cells. A , Cell cycle of A2780 and SKOV3 infected by lentiviral vectors with CEP55 knockdown assessed by flow cytometry. B , Expression of cell cycle-related proteins P21 and Cyclin D1 in A2780 and SKOV3 cells infected by lentiviral vectors with CEP55 knockdown was assessed using western blot. C , Cell migration of A2780 and SKOV3 cells infected by lentiviral vectors with CEP55 knockdown was assessed using Transwell assay. D , Cell invasive capacity of A2780 and SKOV3 cells infected by lentiviral vectors with CEP55 knockdown was assessed using Transwell assay. E , Alterations in the skeleton of A2780 and SKOV3 cells infected by lentiviral vectors with CEP55 knockdown assessed using Phalloidin staining. F , Expression of E-cadherin, Vimentin, Snail, ZEB1, Slug, and N-cadherin in A2780 and SKOV3 cells infected by lentiviral vectors with CEP55 knockdown analyzed using western blot. Statistical significance was assessed using ANOVA. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001. All cell experiments were repeated three times

Article Snippet: Human OC cell lines A2780 (CL-0013) and SKOV3 (CL-0215) were procured from Procell (Wuhan, Hubei, China) and cultured in RPMI-1640 medium containing 10% FBS and 1% penicillin/streptomycin in an incubator containing 5% CO 2 at 37 °C.

Techniques: Knockdown, Infection, Flow Cytometry, Expressing, Western Blot, Migration, Transwell Assay, Staining

Journal: Biology Direct

Article Title: ALKBH5 activates CEP55 transcription through m6A demethylation in FOXP2 mRNA and expedites cell cycle entry and EMT in ovarian cancer

doi: 10.1186/s13062-024-00551-5

Figure Lengend Snippet: FOXP2 mediates transcriptional repression of CEP55. A , The intersection of transcription factors regulating CEP55 downloaded from the hTFtarget database, differentially expressed genes in the GSE119054 dataset, and Differential Genes downloaded from the GEPIA database. B , Expression of FOXP2 in ovarian serous cystadenocarcinoma analyzed by the GEPIA database. C , ChIP-seq database analysis of FOXP2 enrichment in the CEP55 promoter. D , Jaspar database analysis of FOXP2 binding sites in the CEP55 promoter. E , RT-qPCR detection of FOXP2 expression in tumor tissues and adjacent tissues of OC patients. F , Correlation between FOXP2 and CEP55 expression in OC tissues ( n = 19) analyzed using Pearson’s correlation analysis. G , Expression of FOXP2 and CEP55 in cells after overexpression of FOXP2 in A2780 and SKOV3 cells detected by RT-qPCR. H , Luciferase activity in A2780 and SKOV3 cells was assessed using a dual-luciferase reporter assay. I , The binding of FOXP2 to the CEP55 promoter was analyzed using ChIP assay. Statistical significance was assessed using an unpaired t-test or ANOVA. ** p < 0.01, *** p < 0.001, **** p < 0.0001. All cell experiments were repeated three times

Article Snippet: Human OC cell lines A2780 (CL-0013) and SKOV3 (CL-0215) were procured from Procell (Wuhan, Hubei, China) and cultured in RPMI-1640 medium containing 10% FBS and 1% penicillin/streptomycin in an incubator containing 5% CO 2 at 37 °C.

Techniques: Expressing, ChIP-sequencing, Binding Assay, Quantitative RT-PCR, Over Expression, Luciferase, Activity Assay, Reporter Assay

Journal: Biology Direct

Article Title: ALKBH5 activates CEP55 transcription through m6A demethylation in FOXP2 mRNA and expedites cell cycle entry and EMT in ovarian cancer

doi: 10.1186/s13062-024-00551-5

Figure Lengend Snippet: FOXP2-mediated transcriptional repression of CEP55 impedes OC cell malignant phenotype. A , CEP55 expression after overexpression of CEP55 in A2780 and SKOV3 cells overexpressing FOXP2 was analyzed using RT-qPCR. B , Colony formation in A2780 and SKOV3 cells overexpressing both FOXP2 and CEP55 was assessed using colony formation assays. C , The proliferative capacity of A2780 and SKOV3 cells overexpressing both FOXP2 and CEP55 was evaluated using EdU staining. D , Apoptosis in A2780 and SKOV3 cells overexpressing both FOXP2 and CEP55 was evaluated using TUNEL assay. E , Cell cycle changes in A2780 and SKOV3 cells overexpressing both FOXP2 and CEP55 were detected by flow cytometry. F - G , The migration and invasion of A2780 and SKOV3 cells overexpressing both FOXP2 and CEP55 was detected by Transwell assay. H , Alterations in the cytoskeleton of A2780 and SKOV3 cells overexpressing both FOXP2 and CEP55 were assessed using Phalloidin staining. I , Expression of EMT-related proteins E-cadherin, Vimentin, Snail, ZEB1, Slug, and N-cadherin in A2780 and SKOV3 cells overexpressing both FOXP2 and CEP55 analyzed using western blot. Statistical significance was assessed using ANOVA. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001. All cell experiments were repeated three times

Article Snippet: Human OC cell lines A2780 (CL-0013) and SKOV3 (CL-0215) were procured from Procell (Wuhan, Hubei, China) and cultured in RPMI-1640 medium containing 10% FBS and 1% penicillin/streptomycin in an incubator containing 5% CO 2 at 37 °C.

Techniques: Expressing, Over Expression, Quantitative RT-PCR, Staining, TUNEL Assay, Flow Cytometry, Migration, Transwell Assay, Western Blot

Journal: Biology Direct

Article Title: ALKBH5 activates CEP55 transcription through m6A demethylation in FOXP2 mRNA and expedites cell cycle entry and EMT in ovarian cancer

doi: 10.1186/s13062-024-00551-5

Figure Lengend Snippet: FOXP2-mediated transcriptional repression of CEP55 impedes OC cell growth and metastasis in vivo. A , Representative images of abdominal tumors formed by SKOV3 cells in mice. B , Total weight of tumor nodules in the abdominal cavity of mice. C , HE staining analysis of metastatic nodules in mouse lung tissue. D , EMT- and cell cycle-related protein expression in abdominal tumors was assessed using western blot analysis. E , The mRNA expression of FOXP2 and CEP55 in abdominal tumors was evaluated using RT-qPCR. Statistical significance was assessed using ANOVA ( n = 6). * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001

Article Snippet: Human OC cell lines A2780 (CL-0013) and SKOV3 (CL-0215) were procured from Procell (Wuhan, Hubei, China) and cultured in RPMI-1640 medium containing 10% FBS and 1% penicillin/streptomycin in an incubator containing 5% CO 2 at 37 °C.

Techniques: In Vivo, Staining, Expressing, Western Blot, Quantitative RT-PCR

Journal: Biology Direct

Article Title: ALKBH5 activates CEP55 transcription through m6A demethylation in FOXP2 mRNA and expedites cell cycle entry and EMT in ovarian cancer

doi: 10.1186/s13062-024-00551-5

Figure Lengend Snippet: ALKBH5-catalyzed demethylation of FOXP2 represses its expression. A , The m6A modification site of FOXP2 was analyzed using the SRAMP tool. B , Volcano plots of m6A modification differences between OC and control tissues in the GSE119168 and GSE196748 datasets. C , The interactions between FOXP2 mRNA and ALKBH5 protein predicted in the RNAINTER database. D , Correlation between ALKBH5 expression in OC and patients’ survival in the Kaplan-Meier Plotter database. E , The m6A modification of FOXP2 in tumor tissues and adjacent tissues ( n = 19) of OC patients was assessed using MeRIP-qPCR. F , Expression of ALKBH5 in tumor tissues and adjacent tissues of OC patients ( n = 19) detected by immunohistochemistry. G , ALKBH5 mRNA expression in A2780 and SKOV3 cells following infection with lentiviral vectors packaged with ALKBH5 knockdown was evaluated using RT-qPCR. H , FOXP2 and CEP55 mRNA expression in A2780 and SKOV3 cells following infection with lentiviral vectors packaged with ALKBH5 knockdown was evaluated using RT-qPCR. I , Effect of knockdown of ALKBH5 on m6A modification of FOXP2 analyzed by MeRIP-qPCR. J , The binding relation between ALKBH5 and FOXP2 mRNA was analyzed using RIP-qPCR. K , The effect of knockdown of ALKBH5 on FOXP2 mRNA stability was evaluated using actinomycin D-mediated RNA half-life assay. L , The effect of knockdown of ALKBH5 on luciferase activity was assessed using a dual-luciferase reporter assay. Statistical significance was assessed using an unpaired t-test or ANOVA. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001. All cell experiments were repeated three times

Article Snippet: Human OC cell lines A2780 (CL-0013) and SKOV3 (CL-0215) were procured from Procell (Wuhan, Hubei, China) and cultured in RPMI-1640 medium containing 10% FBS and 1% penicillin/streptomycin in an incubator containing 5% CO 2 at 37 °C.

Techniques: Expressing, Modification, Control, Immunohistochemistry, Infection, Knockdown, Quantitative RT-PCR, Binding Assay, Luciferase, Activity Assay, Reporter Assay

Journal: Biology Direct

Article Title: ALKBH5 activates CEP55 transcription through m6A demethylation in FOXP2 mRNA and expedites cell cycle entry and EMT in ovarian cancer

doi: 10.1186/s13062-024-00551-5

Figure Lengend Snippet: Knockdown of ALKBH5 leads to cell cycle arrest and inhibits EMT in OC cells. A , Cell cycle changes in A2780 and SKOV3 cells with ALKBH5 knockdown were detected by flow cytometry. B , Cell cycle-related protein expression in A2780 and SKOV3 cells after knockdown of ALKBH5 was assessed using western blot. C - D , The migration and invasion of A2780 and SKOV3 cells after knockdown of ALKBH5 was detected by Transwell assay. E , Alterations in the cytoskeleton of A2780 and SKOV3 cells after knockdown of ALKBH5 were assessed using Phalloidin staining. F , Expression of EMT-related proteins E-cadherin, Vimentin, Snail, ZEB1, Slug, and N-cadherin in A2780 and SKOV3 cells after knockdown of ALKBH5 was analyzed using western blot. Statistical significance was assessed using ANOVA. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001. All cell experiments were repeated three times

Article Snippet: Human OC cell lines A2780 (CL-0013) and SKOV3 (CL-0215) were procured from Procell (Wuhan, Hubei, China) and cultured in RPMI-1640 medium containing 10% FBS and 1% penicillin/streptomycin in an incubator containing 5% CO 2 at 37 °C.

Techniques: Knockdown, Flow Cytometry, Expressing, Western Blot, Migration, Transwell Assay, Staining

Journal: Technology and Health Care

Article Title: Integrating network pharmacology and Mendelian randomization to explore potential targets of matrine against ovarian cancer

doi: 10.3233/THC-231051

Figure Lengend Snippet: Effects of matrine on OC cell lines A2780 and SKOV3 cells. (A) CCK-8 assay CCK-8 detected the cell viability of A2780 and SKOV3 cells. (B) Flow cytometry detected cell apoptosis. (C) Wound healing detected cell migration. (E) Transwell assay detected cell invasion (scale bar = 50 μ m). ** P < 0.01.

Article Snippet: OC cell lines A2780 (#CL-0013, Procell, Wuhan, China) and SKOV3 cells (#CL-0215, Procell) were cultured in RPMI-1640 medium (Gibco, CA, USA) at 37 ∘ C with 5% CO 2 .

Techniques: CCK-8 Assay, Flow Cytometry, Migration, Transwell Assay

Journal: Technology and Health Care

Article Title: Integrating network pharmacology and Mendelian randomization to explore potential targets of matrine against ovarian cancer

doi: 10.3233/THC-231051

Figure Lengend Snippet: Molecular docking site. (A) CCND1-matrine. (B) IL1B-matrine. (C) TP53-matrine. (D) RT-qPCR detected the mRNA expression of CCND1, IL1B, and P53. OC cell line A2780 and SKOV3 cells were treated with matrine. * P < 0.05, ** P < 0.01.

Article Snippet: OC cell lines A2780 (#CL-0013, Procell, Wuhan, China) and SKOV3 cells (#CL-0215, Procell) were cultured in RPMI-1640 medium (Gibco, CA, USA) at 37 ∘ C with 5% CO 2 .

Techniques: Quantitative RT-PCR, Expressing

Journal: Journal of Ovarian Research

Article Title: Shikonin reduces M2 macrophage population in ovarian cancer by repressing exosome production and the exosomal galectin 3-mediated β-catenin activation

doi: 10.1186/s13048-024-01430-3

Figure Lengend Snippet: SK treatment suppresses malignant phenotype of OC cell lines. A , IC50 value of SK to SKOV3 and A2780 cells tested by applying different doses of SK (1 µM, 2 µM, 4 µM, 8 µM, 16 µM, 32 µM, 64 µM, 128 µM, and 256 µM) for 48 h, examined by CCK-8 assay. SKOV3 and A2780 cells were treated with 5 µM SK for 48 h. B , colony formation capacity of SKOV3 and A2780 cells determined by colony formation assay; C , migration ability of cells analyzed by wound healing assay; D , invasion ability of cells determined by Transwell assay; E , apoptosis of cells measured by TUNEL assay. Three biological replicates were performed. Differences were analyzed by two-way ANOVA. * p < 0.05

Article Snippet: OC cell lines SKOV3 (CL-0215) and A2780 (CL-0013), and a human monocytic leukemia cell line THP-1 (CL-0233), were acquired from Procell Life Science & Technology Co., Ltd. (Wuhan, Hubei, China).

Techniques: CCK-8 Assay, Colony Assay, Migration, Wound Healing Assay, Transwell Assay, TUNEL Assay

Journal: Journal of Ovarian Research

Article Title: Shikonin reduces M2 macrophage population in ovarian cancer by repressing exosome production and the exosomal galectin 3-mediated β-catenin activation

doi: 10.1186/s13048-024-01430-3

Figure Lengend Snippet: SK treatment reduces the promoting effect of OC cell-derived exo on M2 polarization of macrophages. Exo from SKOV3 and A2780 cells, either with or without 5 µM SK treatment for 48 h, were collected, which were designated to OC exo and SK OC exo, respectively. A , morphology of isolated OC exo or SK OC exo determined under TEM; B , particle size distribution and concentration of isolated OC exo or SK OC exo examined by NTA; C , protein expression of exo marker proteins CD9, CD63 and CD81 and endoplasmic reticulum marker Calnexin in the isolated OC exo or SK OC exo determined by WB analysis; D , successful uptake of DiO-labeled exo by THP-1 cells observed under the fluorescence microscope. THP-1 cells were stimulated with 150 nM PMA for 24 h to differentiate into M0 macrophages, followed by treatment with PBS, OC exo or SK OC exo. E - F , percentage of CD163- ( E ) and CD206-( F ) positive THP-1 cells examined by flow cytometry; G , production of IL-10 in the culture supernatant of THP-1 cells examined using ELISA; H , mRNA expression of CD163 and CD206 in THP-1 cells determined by RT-qPCR. Three biological replicates were performed. Differences were analyzed by the one-way or two-way ANOVA. * p < 0.05

Article Snippet: OC cell lines SKOV3 (CL-0215) and A2780 (CL-0013), and a human monocytic leukemia cell line THP-1 (CL-0233), were acquired from Procell Life Science & Technology Co., Ltd. (Wuhan, Hubei, China).

Techniques: Derivative Assay, Isolation, Concentration Assay, Expressing, Marker, Labeling, Fluorescence, Microscopy, Flow Cytometry, Enzyme-linked Immunosorbent Assay, Quantitative RT-PCR

Journal: Journal of Ovarian Research

Article Title: Shikonin reduces M2 macrophage population in ovarian cancer by repressing exosome production and the exosomal galectin 3-mediated β-catenin activation

doi: 10.1186/s13048-024-01430-3

Figure Lengend Snippet: SK treatment reduces tumorigenesis of SKOV3 cells in nude mice and reduces M2 macrophage infiltration. SKOV3 cells were subcutaneously injected into CB-17 SCID mice to generate xenograft tumors, followed by OC exo or SK OC exo treatment. A , growth rate within 4 weeks and weight at week 4 of xenograft tumors in mice; B , Ki67 expression in the xenograft tumor tissues examined by IHC; C , expression of CD163 and CD206 in the xenograft tumor tissues examined by IHC; D , GAL3 expression in the xenograft tumor tissues examined by IHC; E , nuclear and cytoplastic β-catenin levels in macrophages isolated from xenograft tumors determined by WB analysis. In each group, n = 6. Differences were compared by the one-way or two-way ANOVA. * p < 0.05

Article Snippet: OC cell lines SKOV3 (CL-0215) and A2780 (CL-0013), and a human monocytic leukemia cell line THP-1 (CL-0233), were acquired from Procell Life Science & Technology Co., Ltd. (Wuhan, Hubei, China).

Techniques: Injection, Expressing, Isolation

Journal: Nature Communications

Article Title: An all-in-one tetrazine reagent for cysteine-selective labeling and bioorthogonal activable prodrug construction

doi: 10.1038/s41467-024-47188-6

Figure Lengend Snippet: A Protocol for visualizing the activation of tumor-targeting peptide conjugates using a TCO trigger. B Time-lapse imaging of RGD-FL release triggered by the bioorthogonal reaction in live SKOV3 cells. Scale bar: 50 μm. C Plot of pixel fluorescence intensity over time indicating the rapid RGD-FL bioorthogonal cleavage process in live cells. D Confocal image showing efficient uptake and intracellular release of RGD-FL. Scale bar: 10 μm. E Live-cell imaging upon treatment using tumor-targeting peptide conjugates RGD-FL of α v β 3 -positive SKOV3 cells and low-α v β 3 expressed MCF-7 cells (control). Blocking studies were performed by pre-treating SKOV3 cells with a 20-fold excess of unmodified RGD before incubation with RGD-FL and TCO. Scale bar: 50 μm. Three times each imaging experiment was repeated independently with similar results.

Article Snippet: Human ovarian cancer SKOV3 cells (CL-0215), Human breast cancer MCF−7 cells (CL-0149), Human brain glioma U87 cells (CL-0238), Mouse melanoma B16F10 cells (CL-0319), Human hepatic stellate LX2 cells (CL-0560), Human hepatocellular carcinomas HepG2 cells (CL-0103) and Human breast cancer MDA-MB-231 cells (CL-0150) were kindly provided by Procell Life Science & Technology Co., Ltd. Authentication of all cells was conducted via short tandem repeat (STR) profiling in Procell Life Science & Technology Co., Ltd.

Techniques: Activation Assay, Imaging, Fluorescence, Live Cell Imaging, Control, Blocking Assay, Incubation

Journal: Nature Communications

Article Title: An all-in-one tetrazine reagent for cysteine-selective labeling and bioorthogonal activable prodrug construction

doi: 10.1038/s41467-024-47188-6

Figure Lengend Snippet: A Cytotoxicity of Dox, RGD-Dox, and activable-Dox in B16F10, U87, SKOV3, and LX2 cells ( n = 3 biologically independent samples). Gray and blue lines show cells incubated with RGD-Dox or Dox, respectively, for 48 h. The red lines indicate cells were pre-incubated with different concentrations of RGD-Dox for 2 h followed by incubation with TCO for 48 h in total. The insert table shows calculated IC 50 values (μM) for activable-Dox, Dox, and RGD-Dox against corresponding cells. B Prodrug activation (top), improved cytotoxicity and safety (bottom) by employing a bioorthogonal cleavage reaction of RGD-Dox ( n = 3 biologically independent samples). Data were compared using one-way ANOVA followed by Tukey post-hoc test, **** P < 0.0001. C Fluorescence spectra of RGD-Dox (gray) and Dox (blue) at equal concentrations (2 μM) in PBS (10% DMSO). D Fluorescence semi-quantification of the difference in uptake of RGD-Dox and Dox by B16F10 cells ( n = 3 biologically independent samples). Differences between the two groups were assessed for significance using a two-tailed unpaired Student’s t test, **** P < 0.0001. E Fluorescence imaging analysis of the uptake of RGD-Dox or Dox by B16F10 and LX2 cells. Cells were treated with 10 μM RGD-Dox or Dox for 2 h, then washed three times with PBS. Blocking studies were performed by pre-treatment of cells with a 50-fold excess of unmodified RGD before incubation with RGD-Dox. Scale bar: 100 μm. All data are presented as mean±SD. Source data are provided as a Source Data file.

Article Snippet: Human ovarian cancer SKOV3 cells (CL-0215), Human breast cancer MCF−7 cells (CL-0149), Human brain glioma U87 cells (CL-0238), Mouse melanoma B16F10 cells (CL-0319), Human hepatic stellate LX2 cells (CL-0560), Human hepatocellular carcinomas HepG2 cells (CL-0103) and Human breast cancer MDA-MB-231 cells (CL-0150) were kindly provided by Procell Life Science & Technology Co., Ltd. Authentication of all cells was conducted via short tandem repeat (STR) profiling in Procell Life Science & Technology Co., Ltd.

Techniques: Incubation, Activation Assay, Fluorescence, Two Tailed Test, Imaging, Blocking Assay