Caki 1

Manufactured by Thermo Fisher Scientific

Sourced in United States, China, Switzerland

The Caki-1 is a cell line derived from human renal carcinoma cells. It serves as a model system for studying the biology and behavior of kidney cancer cells in a laboratory setting.

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Lab products found in correlation

77 protocols using caki 1

RCC Cell Line Culture and DUXAP8 Silencing

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RCC cell line CAKI1 and A498 was purchased from the Type Culture Collection of the Chinese Academy of Sciences (Shanghai, China). CAKI1 and A498 cells was cultured in Dulbecco's modified Eagle's medium (Invitrogen, Carlsbad, CA) with 10% fetal bovine serum (Invitrogen, shanghai, China), 100 U/ml penicillin and streptomycin (Invitrogen), at 37°C with 5% CO2. The DUXAP8 and negative control siRNAs (Invitrogen, Carlsbad, CA) were transfected into CAKI1 and A498 cells using RNAiMAX (Invitrogen) according to the manufacturer's instructions. 48 hours after transfection, the CAKI1 and A498 cells were harvested for RNA extraction. The DUXAP8 siRNA sequences are: siRNA 1#,5′-AAGATAAAGGTGGTTTCCACAAGAA-3′ siRNA 2#, 5′- CAGCATACTTCAAATTCACAGCAAA-3′

Xu X., Xu Y., Shi C., Wang B., Yu X., Zou Y, & Hu T. (2017). A genome-wide comprehensively analyses of long noncoding RNA profiling and metastasis associated lncRNAs in renal cell carcinoma. Oncotarget, 8(50), 87773-87781.

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Human Clear-Cell Renal Cancer Cell Culture

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We obtained human clear-cell renal cancer cells (786-O, 769-P, ACHN, and Caki-1) from the American Type Culture Collection. All these cells, except Caki-1, were incubated in the RPIM 1640 (Invitrogen, CA, USA) with 10% FBS and 1% penicillin/streptomycin at 37 °C in a humidified 5% CO₂ atmosphere, while Caki-1 cells were maintained in 5 A medium (Invitrogen, CA, USA) containing 10% FBS.
Human anti-IL6 antibody was purchased from R&D Systems (Minneapolis, MN, USA). Anti-STAT3, anti-phosphospecific STAT3 (p-STAT3; Tyr705), anti-MMP2, anti-MMP9, anti-GAPDH, anti-E-cadherin, anti-α-SMA antibodies were purchased from Cell Signaling Technology (Beverly, MA, USA).

Shi Q., Xu R., Song G., Lu H., Xue D., He X, & Xia Y. (2019). GATA3 suppresses human fibroblasts-induced metastasis of clear cell renal cell carcinoma via an anti-IL6/STAT3 mechanism. Cancer Gene Therapy, 27(9), 726-738.

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Culturing Renal Cancer Cell Lines

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Renal cancer cell lines (Caki-1, ACHN, and 769-P) were purchased from the American Type Culture Collection (Rockville, MD). Caki-1 cells were maintained in MEM, ACHN cells in DMEM, and 769-P cells in RPMI medium, all supplemented with 10% fetal bovine serum and 0.3% penicillin-streptomycin (Invitrogen, Carlsbad, CA).

Sato A., Asano T., Isono M., Ito K, & Asano T. (2014). Panobinostat synergizes with bortezomib to induce endoplasmic reticulum stress and ubiquitinated protein accumulation in renal cancer cells. BMC Urology, 14, 71.

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Caki-1 cell line transfection with COLGALT1

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Renal carcinoma cell line Caki-1 and Human renal tubular epithelial cell Hk-2 were ordered at the Shanghai Enzyme-linked Biotechnology Co., Ltd. Caki-1 and Hk-2 were maintained in DMEM and DMEM/F12 medium, respectively, with 10% FBS and 1% penicillin/streptomycin at 37 °C with 5% CO2. KIRC cell line Caki-1 was transfected with negative control (NC) vector and vector of COLGALT1 using lipo3000 reagent (Invitrogen). The vectors of COLGALT1 knockdown and corresponding negative control (NC) were ordered at Gene Pharma. We cultured Caki-1 cells in 6-well plates and used 3ug of plasmid per well for transfection. We conducted further researches after 48 h of cell incubation.

Liu S., Yu Y., Wang Y., Zhu B, & Han B. (2022). COLGALT1 is a potential biomarker for predicting prognosis and immune responses for kidney renal clear cell carcinoma and its mechanisms of ceRNA networks. European Journal of Medical Research, 27, 122.

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Culturing Renal Cell Carcinoma Cell Lines

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786‐O and Caki‐1 RCC cell lines were bought from Cell Bank in the Chinese Academy of Sciences (Shanghai, China). 786‐O cells were cultured in RPMI‐1640 medium (Gibco, USA) with 10% foetal bovine serum (FBS, Gibco, USA) with 1% penicillin/streptomycin (P/S, Hyclone, USA). Caki‐1 cells were cultured in McCoy’5A (Invitrogen) with 10% FBS. RCC4/EV and RCC4/VHL cells were given by Dr JK Cheng in SJTU‐SM, that were cultured in Dulbecco's modified Eagle's medium (DMEM, Hyclone, USA) with 10% FBS with 1% P/S. All cell lines were cultured in 5% CO₂ air at 37°C. There were no signs of mycoplasma contamination for all cell lines.

Zhang Z., Zhang S., Chen H., Mao Y., Li Z., Kong C., Han B., Zhang J., Chen Y., Xue W., Zhai W, & Wang L. (2020). The up‐regulation of NDRG1 by HIF counteracts the cancer‐promoting effect of HIF in VHL‐deficient clear cell renal cell carcinoma. Cell Proliferation, 53(7), e12853.

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Cell Line Maintenance for Renal Cancer

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Human clear cell type RCC cell lines Caki-1 and Caki-2 were purchased from the American Type Culture Collection. Caki-1 and Caki-2 were maintained with McCoy’s 5A medium (Invitrogen) containing 10% fetal bovine serum (FBS). The other human RCC cell lines, UMRC-3 and UMRC-6, were kindly gifted by Dr. P. Black (Vancouver Prostate Centre, UBC). UMRC-3 and UMRC-6 were maintained in MEM medium (Invitrogen) containing 10% FBS and L-glutamine. The human renal proximal tubular epithelial cell line HK-2 was kindly provided by Dr. C. Du (Vancouver Prostate Centre, UBC). HK-2 cells were cultured in DMEM/Ham’s F12 (Invitrogen) supplemented with 10% FBS and glutamine. Immortalized human umbilical vascular endothelial cells (HUVECs) were obtained from Dr. C. Du and maintained with EBM-2 medium (Lonza) containing EGM-2 SingleQuots (Lonza). All cells were cultured at 37°C in a humid atmosphere with 5% CO₂. Mycoplasma contamination was tested. For all in vitro studies, cell lines were passaged for a maximum of 2 months.

Han K.S., Raven P.A., Frees S., Gust K., Fazli L., Ettinger S., Hong S.J., Kollmannsberger C., Gleave M.E, & So A.I. (2015). Cellular Adaptation to VEGF-Targeted Antiangiogenic Therapy Induces Evasive Resistance by Overproduction of Alternative Endothelial Cell Growth Factors in Renal Cell Carcinoma. Neoplasia (New York, N.Y.), 17(11), 805-816.

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Cell Line Generation and Protein Expression

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All cancer cells (Caki-1, A549, MDA-MB231, and HCT116) and TCMK-1 cells were obtained from American Type Culture Collection (Manassas, VA, USA). Cells were grown in an appropriate medium supplemented with 10% fetal bovine serum (FBS) (Welgene, Gyeongsan, Korea), 1% penicillin–streptomycin and 100 μg/mL gentamycin (Thermo Fisher Scientific, Waltham, MA, USA). For constructing stable cell lines, Caki-1 cells were transfected using Lipofectamine^TM 2000 (Invitrogen, Carlsbad, CA, USA) with the pcDNA3.1(+)/c-FLIP or pcDNA3.1(+) vector plasmids. These plasmids were transduced for 24 h and cells were selected by 700 μg/mL G418 (Invitrogen, Carlsbad, CA, USA). For knockdown of the gene by siRNA, Lipofectamine^® RNAiMAX Reagent (Invitrogen, Carlsbad, CA, USA) was used in Caki-1 cells. Immunoblot analysis was performed to examine protein expression [27 (link)].

Woo S.M., Min K.J, & Kwon T.K. (2020). Inhibition of Drp1 Sensitizes Cancer Cells to Cisplatin-Induced Apoptosis through Transcriptional Inhibition of c-FLIP Expression. Molecules, 25(24), 5793.

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Establishing Stable Cell Lines for Protein Expression

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All cancer cells (Caki-1, ACHN, A549 and Hela) and TCMK-1 cells were obtained from American Type Culture Collection (Manassas, VA, USA). Human mesangial cells (MCs) were purchased from Lonza (Basel, Switzerland). Normal human skin fibroblast (HSF) cells were provided by Korea Cell Line Bank (Seoul, Korea). Cells were grown in an appropriate medium supplemented with 10% fetal bovine serum (FBS) (Welgene, Gyeongsan, Korea), 1% penicillin–streptomycin and 100 μg/mL gentamycin (Thermo Fisher Scientific, Waltham, MA, USA). To construct stable cell lines, Caki-1 cells were transfected using Lipofectamine^TM 2000 (Invitrogen, Carlsbad, CA, USA) with pcDNA3.1(+)/Mcl-1, pcDNA3.1(+)/c-FLIP or pcDNA3.1(+) vector plasmids and selected using 700 μg/mL G418 (Invitrogen, Carlsbad, CA, USA). Immunoblot analysis was performed to examine protein expression [31 (link)].

Woo S.M., Min K.J, & Kwon T.K. (2020). Magnolol Enhances the Therapeutic Effects of TRAIL through DR5 Upregulation and Downregulation of c-FLIP and Mcl-1 Proteins in Cancer Cells. Molecules, 25(19), 4591.

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Cell Lines for HLA-A Phenotyping

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T2 (HLA-A*02:01, lymphoblast), Jiyoye (HLA-A32, Burkitt's lymphoma), EB-3 (HLA-A3/Aw32, Burkitt's lymphoma), Cercopithecus aethiops-derived COS7 and A498 (HLA-A*02:01, kidney carcinoma) cells were purchased from the American Type Culture Collection (Rockville, MD). PSCCA0922 (HLA-A*02:06/A*31:01, a B cell line) was provided by the Health Science Research Resources Bank (Osaka, Japan). Caki-1 (HLA-A*24:02/A*23:01, renal clear cell carcinoma) cells were provided by the Cell Resource Center for Biomedical Research Institute of Development, Aging and Cancer at Tohoku University. The HIG2 expression in A498 and Caki-1 cells was confirmed by a Western blotting analysis [24] (link). T2, Jiyoye, EB-3 and PSCCA0922 cells were maintained in RPMI1640 (Invitrogen, Carlsbad, CA), A498 and Caki-1 cells were maintained in EMEM (Invitrogen) and COS7 cells were maintained in DMEM (Invitrogen). Each medium was supplemented with 10% fetal bovine serum (GEMINI Bio-Products, West Sacramento, CA) and 1% antibiotic solution (Sigma-Aldrich, ST. Louis, MO).

Yoshimura S., Tsunoda T., Osawa R., Harada M., Watanabe T., Hikichi T., Katsuda M., Miyazawa M., Tani M., Iwahashi M., Takeda K., Katagiri T., Nakamura Y, & Yamaue H. (2014). Identification of an HLA-A2-Restricted Epitope Peptide Derived from Hypoxia-Inducible Protein 2 (HIG2). PLoS ONE, 9(1), e85267.

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Renal Carcinoma Cell Lines and LINC00460 Modulation

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Human renal carcinoma cell lines 786-O, A-498, Caki-1, 769-P, and normal renal epithelial cells (HK-2) were all purchased from American Type Culture Collection (ATCC, Manassas, USA). Given the outstanding expression of LINC00460, 786-O and/or Caki-1 cells were used for further experiments. 786-O, ACHN, Caki-1, and 769-P cells were all cultured in RPMI-1640 medium (HyClone, Logan, UT, USA) supplemented with 10% FBS (HyClone). HK-2 cells were grown in keratinocyte serum-free medium (K-SFM; Invitrogen, USA) supplemented with 10% FBS (HyClone). All cells were routinely cultured in a humidified circumstance with 5% CO₂ at 37°C.
Small interfering RNAs (siRNAs) designed for LINC00460 (si-LINC00460) and siRNA negative control (si-NC) were obtained from RiboBio Co., Ltd. (Guangzhou, Guangdong, China), as well as the wildtype LINC00460 (WT-LINC00460) and mutant LINC00460 (MUT-LINC00460) and the corresponding negative control (anti-miR-NC). The Recombinant plasmid(s) were then transfected into 786-O cells or Caki-1 cells in the use of Lipofectamine 3000 (Invitrogen, Thermo Fisher Scientific, Inc., Waltham, MA, USA).

Zhang S., Zhang F., Niu Y, & Yu S. (2021). Aberration of lncRNA LINC00460 is a Promising Prognosis Factor and Associated with Progression of Clear Cell Renal Cell Carcinoma. Cancer Management and Research, 13, 6489-6497.

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