A2780

Manufactured by Keygen Biotech

Sourced in China

The A2780 is a cell line commonly used in cancer research. It is an ovarian carcinoma cell line derived from an untreated patient. The A2780 cell line is widely used in the evaluation of anticancer drugs and the study of ovarian cancer biology.

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30 protocols using a2780

Cell Line Procurement and Cultivation

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The ovarian cancer cell lines SKOV3, A2780, and HO8910 were purchased from KeyGEN Biotech (Nanjing, China); 293T cells were purchased from the National Collection of Authenticated Cell Cultures (Shanghai, China); immortalized ovarian surface epithelial cells 80 (IOSE80) were a gift from Dr Li Jing in Nanjing Medical University. All cell lines were verified by STR profiling. SKOV3 cells were cultured in McCoy’s 5A medium (KeyGEN Biotech) supplemented with 10% fetal bovine serum (FBS; Thermo Fisher Scientific, Waltham, USA), 100 ng/mL streptomycin, and 100 U/mL penicillin (Thermo Fisher Scientific). A2780, HO8910, and 293T cells were cultured in high-glucose Dulbecco’s modified Eagle’s medium (DMEM; KeyGEN Biotech) supplemented with 10% FBS, 100 ng/mL streptomycin, and 100 U/mL penicillin. IOSE80 cells were cultured in RPMI-1640 supplemented with 10% FBS, 100 ng/mL streptomycin, and 100 U/mL penicillin. All cells were cultured at 37°C in an incubator with a humidified atmosphere containing 5% CO
₂.

Zhang M., Sun Y., Xu H., Shi Y., Shen R., Teng F., Xu J, & Jia X. (2023). Circular RNA hsa_circ_0007444 inhibits ovarian cancer progression through miR-23a-3p/DICER1 axis: hsa_circ_0007444/miR-23a-3p/DICER1 in ovarian cancer. Acta Biochimica et Biophysica Sinica, 55(4), 574-586.

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Ovarian Cancer Cell Culture Protocol

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The human OC cell line A2780, a serous cystadenocarcinoma line, was obtained from KeyGEN BioTECH (China), and the human OC cell line A2780/DDP was generated by our laboratory. Both A2780 and A2780/DDP cells were maintained in RPMI-1640 medium (HyClone, USA) supplemented with 10% foetal bovine serum (Gibco, USA) and 1% antibiotic–antimycotic solution (Gibco, USA). Cells were cultured at an optimum temperature of 37 °C and under humidified conditions in the presence of 5% CO₂.

Wang H., Liu L., Liu Q., Zheng J., Zheng Q., Chen Y., Xia H., Wu Q, & Sun Y. (2023). Identification of upregulated exosomal miRNAs between A2780 and A2780/DDP human ovarian cancer cells by high-throughput sequencing. Journal of Ovarian Research, 16, 94.

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Culturing Human Ovarian Cancer Cell Lines

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The human OC cell line (SK-OV-3) was purchased from the ATCC (American Type Culture Collection, Manassas, VA, USA) and grown in McCoy's 5A medium (Invitrogen, Carlsbad, CA, USA) containing 10% fetal bovine serum (FBS). Another cell line (A2780) was purchased from KeyGEN (Nanjing, China) and maintained in RPMI 1640 medium supplemented with 10% FBS. The two cell lines were cultured in 5% CO₂ at 37 °C.

Zhang S., Ke X., Zeng S., Wu M., Lou J., Wu L., Huang P., Huang L., Wang F, & Pan S. (2015). Analysis of CD8+ Treg cells in patients with ovarian cancer: a possible mechanism for immune impairment. Cellular and Molecular Immunology, 12(5), 580-591.

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Cell Culture of Ovarian Cancer Lines

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The human ovarian cancer cell lines SKOV3 and A2780 were purchased from Shanghai Zhong Qiao Xin Zhou Biotechnology Co., Ltd. The HEK293 human embryonic kidney cell line (HEK293T) was a kind gift from Professor Chenbo Ji (Nanjing Medical University, Nanjing, China). The SKOV3 cells were cultured in McCoy's 5a medium (Gibco, Grand Island, NY, USA) supplemented with 10% fetal bovine serum (FBS). The A2780 and HEK293T cells were cultured in Dulbecco's modified Eagle's medium (DMEM, KeyGEN BioTECH, Nanjing, China) supplemented with 10% FBS. All cell lines were maintained in a humidified atmosphere containing 5% CO₂ at 37°C.

Tan X., Shao Y., Teng Y., Liu S., Li W., Xue L., Cao Y., Sun C., Zhang J., Han J., Wu X., Xu H, & Xie K. (2021). The Cancer-Testis Long Non-coding RNA PCAT6 Facilitates the Malignant Phenotype of Ovarian Cancer by Sponging miR-143-3p. Frontiers in Cell and Developmental Biology, 9, 593677.

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Ovarian Cancer Cell Regulation by miR-27b-5p

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Ovarian cancer cells (SKOV3, Caov-3, A2780 and OVCAR3) and normal ovarian cell line, HOSEpiC were obtained from Nanjing KeyGen Biotech (Nanjing, Jiangsu, China). Cell lines were maintained in RPMI-1640 medium supplement with 10% FBS (Thermo Fisher Scienti c), 100 µg/ml streptomycin and 100 µg/ml penicillin in 5% CO 2 at 37 °C. MiR-27b-5p mimic or miRNA negative control (NC) mimic was by synthesized by GenePharma (Shanghai, china). MiR-27b-5p stable transfected cells were constructed by using over-expressing miR-27b-5p lentivirus carrier (Vigenebio). CXCL1 cDNA sequences were cloned into a pCDNA3.1 vector (Thermo Fisher Scienti c). MiR-10a-5p combination with pCDNA3.1 vector carrying CXCL1 was transfected into SKOV3 or A2780 cell using Lipofectamine 3000 (Thermo Fisher Scienti c).

Qin S., & Liu M.W. (2020). Tumor-suppressor miRNA-27b-5p regulates the growth and metastatic behaviors of ovarian carcinoma cells by targeting CXCL1.

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Ovarian Cancer Cell Lines: SPP1, ITGB1, FAK, AKT Silencing

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Human ovarian cancer cells lines SKOV3 and A2780 were purchased from Jiangsu KeyGEN BioTECH Corp., Ltd. The cells were cultured in RPM1 1640 medium. The medium was supplemented with 10% fetal bovine serum and 1% penicillin/streptomycin in an atmosphere of 5% carbon dioxide at 37°C. The siRNA sequences were obtained from Guangzhou RiboBio Co., Ltd (Guangzhou, China). The following sequences were targeted for human SPP1, ITGB1, FAK, and AKT siRNA, respectively. SPP1-1: 5′-GGAUGAUAUGGAUGAUGAAGA-3′; SPP1-2: 5′-GAACGACUCUGAUGAUGUAGA-3′; ITGB1: 5′-GGUUACUCUUGUCAGCUAAGG-3′; FAK: 5′-CGAUUAUAUGUUAGAGAUAGC-3′; AKT: 5′-UGUGUAUUAUGUUGUUCAAAU-3′; and negative control (NC) siRNA: 5′-UUCUUCGAAGGUGUCACGUTT-3′. Lentiviral vector expressing shRNA targeting SPP1 (named LV3-shPP1-1 and LV3-shPP1-2) and SPP1-lentiviral expression vector (named LV5-SPP1) were provided by Guangzhou RiboBio Co., Ltd. miR-181a mimics were synthesized at Guangzhou RiboBio Co., Ltd.

Zeng B., Zhou M., Wu H, & Xiong Z. (2018). SPP1 promotes ovarian cancer progression via Integrin β1/FAK/AKT signaling pathway. OncoTargets and therapy, 11, 1333-1343.

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Characterization of Drug-Resistant Cancer Cells

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All cells used in this study were human cancer cells. Ovarian cancer cell line A2780 and its paclitaxel (PTX)-resistant counterpart A2780/PTX (Keygen Biotech, Nanjing, Jiangsu, China), breast cancer cell line MCF-7 (American Type Culture Collection, ATCC, Manassas, VA, USA), its PTX-resistant counterpart MCF-7/PTX and DDP-resistant counterpart MCF-7/DDP (Meixuan Biotech, Shanghai, China) were cultured in DMEM (Gibco, Invitrogen, Carlsbad, CA, USA) supplemented with 10% fetal bovine serum (FBS, Invitrogen). Lung carcinoma cancer cell line A549 (ATCC) and its PTX-resistant counterpart A549/PTX (ATCC), myelogenous leukemia cell line K562 (Keygen Biotech), its doxorubicin (ADR)-resistant counterpart K562/ADR (Keygen Biotech), PTX-resistant counterpart K562/PTX and DDP-resistant counterpart K562/DDP (Meixuan Biotech) were cultured in RPMI-1640 medium (Gibco) supplemented with 10% FBS. The pH value of the culture medium was determined by a pH meter.

Xu X., Wang C., Zhang P., Gao X., Guan W., Wang F., Li X., Yuan J., Dou H, & Xu G. (2022). Enhanced Intracellular Reactive Oxygen Species by Photodynamic Therapy Effectively Promotes Chemoresistant Cell Death. International Journal of Biological Sciences, 18(1), 374-385.

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Characterization of Ovarian and Cancer Cell Lines

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The human epithelial ovarian cancer cell line A2780 was obtained from NANJING KEYGEN BIOTECH CO., LTD, China. SKOV3 cells, human breast cancer cell line (MCF-7, MBA-MD-231), human cervical cancer cell line (Hela), Human leukemia monocyte THP1 cell line were obtained from the Shanghai Institute of Cell Biology at the China Academy of Sciences. The human epithelial ovarian cancer cell lines (A2780 and SKOV3), MCF-7 and THP1 cells were authenticated by Short Tandem Repeat assay (STR). All cells used were passaged less than 3 months after resuscitation. The culture conditions and treatment are described in the Additional file 1: Supplementary methods.

Zhu X., Shen H., Yin X., Yang M., Wei H., Chen Q., Feng F., Liu Y., Xu W, & Li Y. (2019). Macrophages derived exosomes deliver miR-223 to epithelial ovarian cancer cells to elicit a chemoresistant phenotype. Journal of Experimental & Clinical Cancer Research : CR, 38, 81.

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Establishment and Characterization of Cancer Cell Lines

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All cell lines used were of human origin. A549 (non-small cell lung cancer) and MCF-7 (breast cancer) cell lines were purchased from Cell Resource Center of Shanghai Institute for Biological Sciences (Chinese Academy of Sciences, Shanghai, China). A549T (Taxol-resistant A549 subline) and MCF-7R (Adriamycin-resistant MCF-7 subline) cell lines were from Shanghai Aiyan Biological Technology Co. Ltd (Shanghai, China). HCT-8 (colon carcinoma), A2780 (ovarian cancer), HCT-8T (Taxol-resistant HCT-8 subline) and A2780T (Taxol-resistant A2780 subline) were from Nanjing KeyGEN BioTECH Co. Ltd (Nanjing, China). PC-3 (prostate cancer) was from Typical Culture Preservation Commission Cell Bank, Chinese Academy of Sciences (Shanghai, China).
All cell lines were routinely cultured at 37 °C in humidified atmosphere with 5% CO₂. Unless stated otherwise, the growth medium used for A549, A549T, MCF-7R, HCT-8, HCT-8T, PC-3 and A2780T cells was RPMI-1640 (Corning) containing 10% fetal bovine serum (FBS, Gibco) and 1% penicillin/streptomycin (Corning). MCF-7 and A2780 cells were propagated in Dulbecco’s Modified Eagle Medium (DMEM, Corning) with 10% fetal bovine serum (FBS, Gibco) and 1% penicillin/streptomycin (Corning). For comparison, some experiments replaced FBS with other serums (calf serum, bovine serum, goat serum, horse serum or human serum) and the same concentration.

Wang L., Liu Y., Qi C., Shen L., Wang J., Liu X., Zhang N., Bing T, & Shangguan D. (2018). Oxidative degradation of polyamines by serum supplement causes cytotoxicity on cultured cells. Scientific Reports, 8, 10384.

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Culturing Ovarian Cancer Cell Lines

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Five OC cell lines (SW626, SK-OV-3, TOV-112D, A2780, and OVcar3) and a normal ovarian epithelial cell line (IOSE-80) were purchased from Nanjing KeyGen Biotech (Nanjing, China). All the cell lines were initially stored in liquid nitrogen and subsequently allowed to recover in a complete DMEM medium (Thermo Fisher Scientific, Waltham, MA, USA) containing 10% fetal bovine serum, 10,000 UI/mL penicillin, and 10 mg/mL streptomycin prior to use. The cells then were cultured in a humidified 37°C incubator containing 5% carbon dioxide.

Kou X., Ding H., Li L, & Chao H. (2021). Hsa-miR-105-1 Regulates Cisplatin-Resistance in Ovarian Carcinoma Cells by Targeting ANXA9. Analytical Cellular Pathology (Amsterdam), 2021, 6662486.

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