ovarian cancer cell line ov90  (ATCC)

Journal: Cell Death & Disease

Article Title: Splice-switching of the oncogenic BCS1L isoform suppresses ovarian cancer progression by disrupting mitochondrial function

doi: 10.1038/s41419-026-08495-6

Figure Lengend Snippet: A Venn diagram of 53 BCS1L -bound splicing factors from the BCS1L RNA pulldown-MS in A2780 cells and 24 genes regulating BCS1L splicing derived from the TCGA-OV datasets (Pearson’s R > 0.15, p < 0.001). B Volcano plots of differentially expressed BCS1L -bound splicing factors between the TCGA-OV ( n = 374) dataset and the normal tissue in GTEx-ovary ( n = 180) datasets. |Pearson R | > 0.1 and p -value > 0.05 were considered significant. Positive correlations are shown in red, and negative correlations are shown in blue. C Relative BCS1L-L/BCS1L-S transcript expression was measured by qPCR in A2780 cells transfected with siRNAs targeting YBX1 , PUF60 , HSPA8 , USP39 , HNRNPF , SF3B4 , SNRPC , and SNRNP40 or a negative control for 48 h. D Sashimi plots of the alternative splicing pattern and USP39 direct binding sites in BCS1L were created with IGV using RNA-seq and RIP-seq data in A2780 cells. The light blue region indicates the alternative exon and the USP39-binding sites. E Semi-quantitative RT-PCR was performed to validate alternative splicing events in A2780 cells with USP39 knockdown using isoform-specific primers. F The relative expression ratio of BCS1L-L/BCS1L-S was analyzed in A2780 cells with USP39 depletion using isoform-specific primers. G Schematic diagram of the BCS1L minigene structure and alternative splicing products. H The expression of BCS1L minigene transcripts in 293T cells with USP39 depletion using different USP39 siRNAs as measured by semi-quantitative RT-PCR. I The expression of BCS1L minigene transcripts in 293T cells transfected with USP39 expression plasmids was measured by semi-quantitative RT-PCR. J The interaction between USP39 and BCS1L RNA was validated by RIP-qPCR of A2780 cells overexpressing USP39. U6snRNA served as the positive control. n = 3 biologically independent experiments. K RNA pull-down assay showing the interaction between the BCS1L RNA and USP39 protein in A2780 cells. Androgen receptor 3´-UTR RNA was used as the positive control and poly (A) 25 RNA was used as the negative control. L The protein expression of BCS1L-L and BCS1L-S was determined by western blot in A2780, HEY, and OV90 cells with USP39 knockdown and in OVCAR3 cells with USP39 overexpression. The top band at 48 kDa indicates BCS1L-L, and the bottom band at 34 kDa indicates BCS1L-S. M Representative confocal images of control A2780 cells and A2780 cells with USP39 knockdown showing co-localization of BCS1L (green) and COX4 (red). DAPI was used to visualize nuclei. Scale bars, 10 µm. Images are representative of at least three independent experiments. Co-localization coefficients, including Pearson correlation coefficient and Mander’s coefficient were quantified by Image J ( n = 50 cells for control, n = 50 cells for USP39 knockdown). The p -value ( C , F , J , M ) was obtained by Student’s unpaired t -test and and the results are presented as the mean ± SD. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: Human ovarian cancer cell lines OV90, SKOV3, OVCAR3, OVCAR8, and CAOV3 were purchased from the American Type Culture Collection.

Techniques: Derivative Assay, Expressing, Transfection, Negative Control, Alternative Splicing, Binding Assay, RNA Sequencing, Quantitative RT-PCR, Knockdown, Positive Control, Pull Down Assay, Western Blot, Over Expression, Control

Journal: Journal for Immunotherapy of Cancer

Article Title: Universal off-the-shelf combination immunotherapy using oncolytic viruses to redirect T cell engagers to target solid tumors

doi: 10.1136/jitc-2024-011051

Figure Lengend Snippet: OV effectively delivers BCMAt to solid tumors and redirects activation and cytotoxicity of BCMA-CAR T cells and human T cells in the presence of BCMA-TCE in vitro. ( a ) Illustration of vaccinia OV with BCMAt incorporated under the control of the synthetic early promoter inserted into the J2R locus and replacing the tk gene (OVBCMAt). ( b ) Quantification of BCMAt expression on various solid tumor cell lines 24 (left) and 48 hours (right) after exposure to indicated MOI of OVBCMAt. n=2 per condition and each experiment was repeated two times. ( c–e ) Quantification of CD137 expression (left), tumor cell killing (middle), and BCMAt expression (right) assessed by flow cytometry after 24 hours of ( c ) MDA-MB-468, ( d ) SNU-16, or ( e ) OV90 tumor cells co-cultured with or without BCMA-TCE in the presence of human T cells or BCMA-CAR T cells and indicated MOI of OVBCMAt. n=2 per condition and each experiment was repeated three times. P values indicate differences between BCMA-TCE groups with PBMCs and PBMCs only determined using unpaired Student’s t-test (*p<0.05, **p<0.01, and ***p<0.005; ns=not significant). BCMAt, truncated B cell maturation antigen; CAR, chimeric antigen receptor; MOI, multiplicity of infection; OV, oncolytic virus, OVBCMAt, OV carrying the BCMAt-encoding gene; PBMC, peripheral blood mononuclear cell; TCE, T cell engager; tk, thymidine kinase; UTD, untransduced.

Article Snippet: Human ovarian cancer cell line OV90 (ATCC CRL-11732) was cultured in 1:1 vol of MCDB 105 medium (Sigma-Aldrich) and medium 199 (Gibco) containing 20% FBS and 1× AA.

Techniques: Activation Assay, In Vitro, Control, Expressing, Flow Cytometry, Cell Culture, Infection, Virus

Journal: Frontiers in bioscience (Landmark edition)

Article Title: N6-methyladenosine Methyltransferase METTL3 Enhances PTGER2 Expression to Increase Ovarian Cancer Stemness and Chemoresistance.

doi: 10.31083/j.fbl2809199

Figure Lengend Snippet: Fig. 2. METTL3 is responsible for m6A modification of PTGER2 mRNA. (A) quantitative PCR (qPCR) analysis determining the efficiency of METTL3 knockdown in OV90 and SKOV3 cells. (B) qPCR analysis showing the effect of METTL3 knockdown on

Article Snippet: Ovarian cancer cell lines OV90 and SKOV3with short tandem repeat verification were purchased from American Type Culture Collection (Manassas, VA, USA) and cultured in Dulbecco’s Modified Eagle Medium (DMEM) harboring 10% fetal bovine serum (FBS) at 37 °C with 5% CO2.

Techniques: Modification, Real-time Polymerase Chain Reaction, Knockdown