Sk ut 1

Manufactured by American Type Culture Collection

Sourced in United States, United Kingdom

The SK-UT-1 is a cell line derived from a uterine sarcoma. It is maintained in cell culture and can be used for research purposes.

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24 protocols using sk ut 1

Sarcoma and Rhabdoid Tumor Cell Lines

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Sarcoma cell lines U2OS (osteosarcoma), MES-SA (uterine sarcoma), and SKUT1 (leiomyosarcoma) were obtained from ATCC (Manassas, VA), as was rhabdoid tumor cell line A204 (rhabdoid tumors bear a SMARCB1 mutation and are mostly found in small children). Cells were cultured with 10% fetal bovine serum and 2% antibiotics (Penicillin-Streptomycin) by following the manufacturer’s recommendations for growth media.

Jiang L., Tolani B., Yeh C.C., Fan Y., Reza J.A., Horvai A.E., Xia E., Kratz J.R., Jablons D.M, & Mann M.J. (2019). Differential gene expression identifies KRT7 and MUC1 as potential metastasis-specific targets in sarcoma. Cancer Management and Research, 11, 8209-8218.

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Leiomyosarcoma Cell Line Cytotoxicity Assay

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The following leiomyosarcoma cell lines were purchased from the ATCC: SK-LMS1 (ATCC^® HTB-88^™, leiomyosarcoma of the vulva), SK-UT1 (ATCC^® HTB114^™, uterine leiomyosarcoma), and MES-SA (ATCC^® CRL-1976^™, poorly differentiated uterine sarcoma). CellTox^™ Green Cytotoxicity Assay (cat. No. G8741), CellTiter96 Cell Proliferation Assay (cat. No. G3580) and Caspase Glo^® Assay (cat. No. G8090) were purchased from Promega. 5-azacitidine (cat. No. A2385) and 5-aza-2′-deoxycytidine (cat. No. A3656) were purchased from Sigma-Aldrich. Guadecitabine (SGI 110) was supplied by ASTEX pharmaceuticals. NOD/SCID mice were purchased from (Jackson Laboratories).

De Carvalho Fischer C., Hu Y., Morreale M., Lin W.Y., Wali A., Thakar M., Karunasena E., Sen R., Cai Y., Murphy L., Zahnow C.A., Keer H., Thakar M, & Ahuja N. (2018). Treatment with epigenetic agents profoundly inhibits tumor growth in leiomyosarcoma. Oncotarget, 9(27), 19379-19395.

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Smooth Muscle Differentiation of BM-MSCs

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Human BM-MSCs were obtained from the Texas University Institute of Regenerative Medicine MSC Distribution Program (http://medicine.tamhsc.edu/irm/msc-distribution.html) and cultured as previously described [14 (link)]. SK-LMS1, SK-UT1 and 293T cell lines were purchased from ATCC (Manassas, VA, USA) and cultured in IMDM (LMS cells) or DMEM (293T) (Invitrogen, Carlsbad, CA, USA), supplemented with 10% fetal bovine serum (Cell Gro, Manassas, VA) and 100U/ml penicillin/streptomycin. Smooth muscle differentiation of BM-MSCs using TxA2 for myogenic induction was performed as previously described [14 (link), 28 (link)]. Induction was performed by media supplementation with 1.0 μM of the TXA2 analog U46619 (Enzo Life Sciences, Farmingdale, NY, USA).

Danielson L.S., Guijarro M.V., Menendez S., Higgins B., Sun Q., Mittal K., Popiolek D.A., Overholtzer M., Palmer G.D, & Hernando E. (2023). MiR-130b modulates the invasive, migratory, and metastatic behavior of leiomyosarcoma. PLOS ONE, 18(1), e0278844.

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Sarcoma Cell Line Cultivation and Maintenance

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The GCT, SKLMS1, SKUT1, SJCRH30, and sNF96.2 sarcoma cell lines were purchased from ATCC (American Type Culture Collection). After thawing and expansion, 10 vials of each were frozen. Upon thawing, they were maintained in culture for no longer than 2 months. HeLa cells were also obtained from ATCC.
Cells were cultured in high-glucose (4.5 g/L) DMEM (Dulbecco’s modified Eagle’s medium) containing L-glutamine (4 mM) and L-pyruvate (5 mM) or in RPMI 1640 medium (SJCRH30). The complete growth medium was made by adding 10% FBS (fetal bovine serum) and antibiotics (penicillin (100 U/mL), streptomycin (100 µg/mL)). Cell culture reagents were purchased from Thermo Fisher (Madrid, Spain). All cell lines were cultured at 37 °C in a humidified atmosphere in the presence of 5% CO₂ and 95% air. Mycoplasma testing was routinely performed.

Sánchez-Fdez A., Matilla-Almazán S., Del Carmen S., Abad M., Arconada-Luque E., Jiménez-Suárez J., Chinchilla-Tábora L.M., Ruíz-Hidalgo M.J., Sánchez-Prieto R., Pandiella A, & Esparís-Ogando A. (2023). Etiopathogenic role of ERK5 signaling in sarcoma: prognostic and therapeutic implications. Experimental & Molecular Medicine, 55(6), 1247-1257.

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Sarcoma Cell Lines for Gene Expression

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The following STS cell lines were used for ERCC1, ERCC5 and CUL4A gene expression analysis: Liposarcoma cell lines 93T449 (ATCC^® CRL-3043™; ATCC, Old Town Manassas, VA, USA) and SW872 (ATCC^® HTB-92™; ATCC, Manassas, VA, USA); leiomyosarcoma primary cell lines AA (kindly provided by Dr. Amancio Carnero of the Institute of Biomedicine of Seville, CSIC, US, HUVR; Seville, Spain) and CP0024 (established in Martin-Broto laboratory); SW982 (ATCC^® HTB-93™; ATCC) synovial sarcoma cell line; fibrosarcoma cell line HT-1080 (ATCC^® CCL-121™; ATCC) and uterine leiomyosarcoma cell line SK-UT-1 (ATCC^® HTB-114™; ATCC).
HT-1080 and AA cell line were maintained in F-10 medium (Gibco^TM, Thermo Fischer Scientific, Waltham, MA, USA), 93T449 and CP0024 were cultured in RPMI cell medium (Gibco^TM), SK-UT-1 was maintained in DMEM cell culture medium (Gibco^TM) and both SW872 and SW982 were cultured in Leibovitz’s L-15 Medium (Gibco^TM). All the cell culture mediums were supplemented with 10% FBS, and 100 units/mL penicillin (PAA) and 100 μg/mL streptomycin. Cells were checked routinely and test for contamination by Mycoplasma or fungi. All the cells lines were discarded after 2 months and new lines obtained from frozen stocks.

Moura D.S., Sanchez-Bustos P., Fernandez-Serra A., Lopez-Alvarez M., Mondaza-Hernandez J.L., Blanco-Alcaina E., Gavilan-Naranjo A., Martinez-Delgado P., Lacerenza S., Santos-Fernandez P., Carrasco-Garcia I., Hidalgo-Rios S., Gutierrez A., Ramos R., Hindi N., Taron M., Lopez-Guerrero J.A, & Martin-Broto J. (2020). CUL4A, ERCC5, and ERCC1 as Predictive Factors for Trabectedin Efficacy in Advanced Soft Tissue Sarcomas (STS): A Spanish Group for Sarcoma Research (GEIS) Study. Cancers, 12(5), 1128.

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Cell Line Characterization and Validation

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Cell lines (SK-LMS-1, SK-UT-1, HTB-93, HT-1080, SK-MEL-2, AS-Pc-1, MiaPaCa-2, MNNG, RDES, and RD HPAC) were all purchased from ATCC. SYO-1 and FUJI were kindly provided by Dr. Akira Kawai (National Cancer Centre Hospital, Tokyo, Japan) and Dr. Kazuo Nagashima (Hokkaido University School of Medicine, Sapporo, Japan), LUPI was a gift from Dr. John Pfeiffer (Washington University in St. Louis). RH28 were a gift from Lee J. Helman (USC Children's Hospital, Los Angeles, California). All cell lines were cultured at 37°C in 5% CO₂, in Dulbecco Modified Eagle Medium (DMEM, Life Technologies) supplemented with 10% FBS (Gibco, Thermo Fisher). For experiments, Modified Eagle Medium (MEM; Life Technologies), supplemented with 10% FBS, was used. All LTAT cell lines were cultured in DMEM or MEM with 1 μg/mL ADI-PEG20 (Polaris) until ASS1 expression was detected, and were then continuously cultured in ADI-PEG20–supplemented media. All cell lines other than LUPI, RD, SYO-1, FUJI, RDES, and RH28 are from ATCC and cultured less than 6 months. The SYO-1 and FUJI cell lines were authenticated by confirming the expression of the pathognomic SYT-SSX fusion gene by RT-PCR, LUPI, and RD were authenticated by confirming the expression of the pathognomic EWS-FLI fusion gene by RT-PCR. All cell lines were determined to be Mycoplasma free using the LookOut Mycoplasma PCR Detection kit (Sigma-Aldrich).

Prudner B.C., Rathore R., Robinson A.M., Godec A., Chang S.F., Hawkins W.G., Hirbe A.C, & Van Tine B.A. (2019). Arginine Starvation and Docetaxel Induce c-Myc-Driven hENT1 Surface Expression to Overcome Gemcitabine Resistance in ASS1-Negative Tumors. Clinical cancer research : an official journal of the American Association for Cancer Research, 25(16), 5122-5134.

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Cancer Cell Culture Protocols

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p53 WT cancer cells including A375, IM-9, HepG2, SKCO-1, HeLa, MCF-7, U-87-MG, LNCaP-FGC, NCI-H711, HCT116, U2OS and p53 MT cancer cells including MDA-MB-468, MOLT4, SW837, NCI-H23, P3HR1, PSN1, SKLMS1, SKLU1, SK-UT-1 and SNU-16 lines were procured from ATCC (VA, USA). HEK293T cells were procured from Cell BioLabs Inc. (San Diego, CA) and HEK293FT cells were procured by Thermo Fisher Scientific (MA, USA). Briefly, the cells were cultured in monolayer in respective growth mediums (Dulbecco's modified Eagle's medium (DMEM) or RPMI 1640) supplemented with 10% (v/v) heat-inactivated fetal bovine serum (FBS) and antibiotics (1% penicillin/streptomycin) and incubated under normoxic conditions at 37°C in a humidified atmosphere of 95% air and 5% CO₂.

Madan E., Parker T.M., Pelham C.J., Palma A.M., Peixoto M.L., Nagane M., Chandaria A., Tomás A.R., Canas-Marques R., Henriques V., Galzerano A., Cabral-Teixeira J., Selvendiran K., Kuppusamy P., Carvalho C., Beltran A., Moreno E., Pati U.K, & Gogna R. (2019). HIF-transcribed p53 chaperones HIF-1α. Nucleic Acids Research, 47(19), 10212-10234.

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Silencing SHARPIN in Uterine Sarcoma Cells

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Human uterine sarcoma cell lines SK-UT-1 and MES-SA were obtained from ATCC. MES-SA was cultured in Gibco™ DMEM (Carlsbad, CA), and SK-UT-1 was cultured in Gibco™ MEM supplemented with 10% FBS (Grand Island, NY, USA). SHARPIN siRNA (target sequence 5’-CCCTGAGTGTTCAGCTTCA-3’) and negative-control siRNA (target sequence 5’-GGCTCTAGAAAAGCCTATGC-3’) were transfected into cells (5 µg siRNA, ratio 1:1 duplexes) using ExFect2000 Transfection reagent (Vazyme, Nanjing, China) for 72 h according to the manufacturer’s instructions and were subsequently collected for Cell Counting Kit-8 (CCK-8), Western blot (WB) and colony formation assays.

Chen L., Li J., Wu X, & Zheng Z. (2021). Identification of Somatic Genetic Alterations Using Whole-Exome Sequencing of Uterine Leiomyosarcoma Tumors. Frontiers in Oncology, 11, 687899.

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Characterization of Sarcoma Cell Lines

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Human HT-1080 (Fibrosarcoma), RD (Rhabdomyosarcoma), LPS224 and LPS863 (Liposarcoma), SKLMS1 and SKUT1 (Leiomyosarcoma) and HEK-293T cell lines were purchased from ATCC (Manassas, VA, USA). STS-109 and STS-48 cell lines were derived from human UPS patients. KP230 and KIA cell lines were derived from UPS mouse tumors as described in Eisinger-Mathason et al. (4 (link)). STR analysis was performed at the time of derivation and confirmed in April 2015. Cells were purchased, thawed, and then expanded in the laboratory. Multiple aliquots were frozen down within 10 days of initial resuscitation. For experimental use, aliquots were resuscitated and cultured for up to 20 passages (4–6 wk) before being discarded. Cells were cultured in DMEM with 10% (vol/vol) FBS and 1% penicillin/streptomycin. All cell lines were confirmed to be negative for mycoplasma contamination.

Ye S., Lawlor M., Rivera-Reyes A., Egolf S., Chor S., Pak K., Ciotti G., Lee A., Marino G., Shah J., Niedzwicki D., Weber K., Park P.M., Alam M.Z., Grazioli A., Haldar M., Xu M., Perry J.A., Qi J, & Eisinger-Mathason T.S. (2018). YAP1-mediated suppression of USP31 enhances NF-κB activity to promote sarcomagenesis. Cancer research, 78(10), 2705-2720.

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Characterization of ULMS Cell Lines

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Three ULMS-derived cell lines were used for this study. SKN was purchased from the Japanese Cancer Research Resources Bank, and SK-UT-1 and SK-LMS-1 were purchased from the ATCC. According to the depmap portal (https://depmap.org/portal/), all cell lines contain different missense mutations in TP53. Moreover, SK-UT-1 contains frameshift mutations in RB1 and PTEN, whereas SKN contains a missense mutation in PTEN. SKN cells were maintained in Ham's F12 medium (Sigma-Aldrich) containing 10% FBS (Thermo Fisher Scientific) and antibiotics. SK-UT-1 and SK-LMS-1 cells were maintained in MEM (Nacalai Tesque) containing 10% FBS, 1 mmol/L sodium pyruvate (Thermo Fisher Scientific), and antibiotics. The cell lines tested negative to Mycoplasma contamination and were used between five and 40 passages for the experiments.

Yoshida K., Yokoi A., Yamamoto T., Hayashi Y., Nakayama J., Yokoi T., Yoshida H., Kato T., Kajiyama H, & Yamamoto Y. (2022). Aberrant Activation of Cell-Cycle–Related Kinases and the Potential Therapeutic Impact of PLK1 or CHEK1 Inhibition in Uterine Leiomyosarcoma. Clinical Cancer Research, 28(10), 2147-2159.

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