human ovarian cancer cell lines 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