Human renal cell carcinoma cell lines (786-O, ACHN, and Caki-1) were purchased from American Type Culture Collection (ATCC; Manassas, VA, USA) and maintained in culture media. 786-O cells were maintained in an incubator at 37 °C and 5% CO2 in RPMI 1640 medium (Roswell Park Memorial Institute, Gibco/Life Technologies, Carlsbad, CA, USA) supplemented with 10% fetal bovine serum (FBS). ACHN and Caki-1 cells were maintained in Dulbecco’s modified Eagle’s medium (DMEM; Gibco/Life Technologies) supplemented with 10% FBS and 1% penicillin-streptomycin Mixed Solution (Nacalai Tesque, Kyoto, Japan). Experiments with the majority of cell lines were conducted within 3–6 months and 10 passages of purchase from ATCC, and each cell line was seeded at 2 × 106 cells/dish for subsequent analysis. Sunitinib-resistant RCC cells were generated by growing parental 786-O (786-P) cells serially treated with increasing concentrations of Sunitinib (S1042, Selleck, Houston, TX, USA) up to 10 µM. After continuous culture in complete medium supplemented with 10 µM Sunitinib for >20 passages, these cells were used as Sunitinib-resistant RCC cells (786-R) and maintained in medium containing 10 µM Sunitinib [22 (link),23 (link),24 (link)].
Sunitinib
Manufactured by Selleck Chemicals
Sourced in United States, Germany, China
Sunitinib is a small-molecule inhibitor of multiple receptor tyrosine kinases, including vascular endothelial growth factor receptors (VEGFR) 1, 2, and 3, platelet-derived growth factor receptors (PDGFR) α and β, stem cell factor receptor (KIT), Fms-like tyrosine kinase-3 (FLT3), and the receptor encoded by the RET proto-oncogene.
Automatically generated - may contain errors
Lab products found in correlation
Imatinib, by Selleck Chemicals (14 mentions)
Sorafenib, by Selleck Chemicals (13 mentions)
Dasatinib, by Selleck Chemicals (11 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (10 mentions)
Pazopanib, by Selleck Chemicals (9 mentions)
Axitinib, by Selleck Chemicals (9 mentions)
Erlotinib, by Selleck Chemicals (8 mentions)
Gefitinib, by Selleck Chemicals (7 mentions)
Lapatinib, by Selleck Chemicals (6 mentions)
Ponatinib, by Selleck Chemicals (6 mentions)
Trametinib, by Selleck Chemicals (6 mentions)
Cabozantinib, by Selleck Chemicals (6 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (6 mentions)
Regorafenib, by Selleck Chemicals (5 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (5 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (5 mentions)
Ruxolitinib, by Selleck Chemicals (4 mentions)
Crizotinib, by Selleck Chemicals (4 mentions)
Mk 2206, by Selleck Chemicals (4 mentions)
Everolimus, by Selleck Chemicals (4 mentions)
97 protocols using sunitinib
1
Establishment of Sunitinib-Resistant Renal Cell Carcinoma Cell Lines
2
RIP1-Tag2 Mouse Model for Sunitinib Treatment
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
All animal procedures were performed in accordance with relevant guidelines and regulations and approved by the Stockholm North Ethical Committee on Animal Research, Stockholm, Sweden. The following mouse strains were used: RIP1-Tag2 (RT2)3 (link), Tie2-Cretg/wt16, R26R-Ai3-EYFPfl/wt17, RIP-CreTg/Wt21, R26R-Ai14-Tomatofl/wt17, RIP-VEGF-B25 (link) and maintained on C57Bl/6 J background.
For Sunitinib treatment, Sunitinib (SelleckChem) was dissolved at 80 mg/ml in 100% DMSO (Sigma), allocated and frozen at −21 °C for up to 1 week. Pure DMSO was similarly allocated and frozen. 200 μl DMSO, with Sunitinib or without (vehicle), were freshly mixed with 400 μl PEG400 (Sigma) and 400 μl DPBS (Sigma) and injected subcutaneous within 30 min. Every mouse received 50 μl/day for 14 consecutive days of the Sunitinib solution (treatment) or vehicle (sham treatment).
For Sunitinib treatment, Sunitinib (SelleckChem) was dissolved at 80 mg/ml in 100% DMSO (Sigma), allocated and frozen at −21 °C for up to 1 week. Pure DMSO was similarly allocated and frozen. 200 μl DMSO, with Sunitinib or without (vehicle), were freshly mixed with 400 μl PEG400 (Sigma) and 400 μl DPBS (Sigma) and injected subcutaneous within 30 min. Every mouse received 50 μl/day for 14 consecutive days of the Sunitinib solution (treatment) or vehicle (sham treatment).
3
Establishment of Sunitinib-Resistant OS-RC-2 Cell Line
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The sunitinib-resistant OS-RC-2 cell line was established by continuous exposure to increasing concentrations of sunitinib (Selleck Chemicals) for ~12 weeks. The initial concentration of sunitinib was 1 µM, increased to 2 µM after 4 weeks, to 5 µM after 4 weeks, and maintained at 5 µM for the last 4 weeks.
4
Cell Cycle and Invasion Assays for Targeted Therapies
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
For the treatment of TRZ, each cell was seeded on 6-well plates and after 24 h, TRZ was treated to the cells at 50 μg/mL for 48 h. The cell cycle was analyzed by flow cytometry. For the treatment of ponatinib and sunitinib, each cell (2 × 103 cells/well) was seeded on 96-well plates and after 24 h, ponatinib (Selleck Chemicals, Houston, TX, USA), sunitinib (Selleck Chemicals), or S3I-201 were treated at the indicated concentration for 48 h and for 10 days. In the cell invasion assay, each cell was seeded in the Boyden-chamber inserts with or without 1 μM pontatinib or sunitinib for 24 h. Phosphate-buffered saline was treated as a vehicle control for TRZ treatment and dimethyl sulfoxide was treated as a vehicle control for inhibitors including ponatinib, sunitinib, and S3I-201.
5
Establishment and Characterization of Sunitinib-Resistant Renal Cell Carcinoma Cell Lines
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
RCC cell lines used in this study were obtained from the Cell Bank of the Type Culture Collection of the Chinese Academy of Sciences (Shanghai, China) in 2017. HK-2 cells were maintained in DMEM, High Glucose medium (Gibco, Waltham, MA, USA). A498 and ACHN cells were cultured in Minimum Essential Medium (Gibco). 786-O cells were maintained in RPMI-1640 medium (Gibco). Caki-1cells were maintained in McCoy's 5A Medium (Gibco). The culture media of all cell lines were supplemented with fetal bovine serum (FBS, 10%, Gibco), 1% penicillin/streptomycin (Gibco). RCC cell lines were cultured at 37 °C in 5% Co2. Sunitinib-resistant 786-O and 769-p cell lines (786-O-SR and 769-p-SR) were continuously exposed to increasing dose of Sunitinib (Selleck, USA) for about 12 weeks. The starting dose was 5 μM and this was increased to 10 μM after 4 weeks, to 15 μM after another 4 weeks and continued at 15 μM for the last 4 weeks. The established resistant 786-O and 769-P cell lines were then maintained in DMEM medium with 10% (v/v) FBS and 10 μM Sunitinib.
The cell lines in this study were authenticated by short tandem repeat (STR) profiling and detected for mycoplasma contamination using a Mycoplasma Detection Kit (Selleck Chemicals), and the most recent tests were conducted in September 2018. All cell lines used in this study were cultured within 40 passages.
The cell lines in this study were authenticated by short tandem repeat (STR) profiling and detected for mycoplasma contamination using a Mycoplasma Detection Kit (Selleck Chemicals), and the most recent tests were conducted in September 2018. All cell lines used in this study were cultured within 40 passages.
6
Breast Cancer Cell Line Characterization
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
MCF7, BT549, MDAMB231, MDAMB468, Hs578T, SUM159PT, BT549, and BT20 breast cancer cell lines were obtained from ATCC (Manassas, VA, USA). MCF7 cells were grown in DMEM medium, the other cell lines were grown in RPMI medium, with 10% fetal bovine serum (FBS), penicillin (100 U/mL), and streptomycin (100 µg/mL). Cells were plated 24 h before treatment with different drugs at the indicated concentrations. Recombinant human EGF, heregulin (HRG), IGF1, angiotensin II (Ang II), and angiotensin 1–7 (Ang 1–7) were obtained from Sigma Chemical Co (St. Louis, MO, USA). Recombinant HGF and FGF were from Abcam. Recombinant PDGF-BB was from Gibco. Trastuzumab, gifitinib, lapatinib, sunitinib, OSI-906, and doxorubicin were obtained from Selleck Chemicals. The PRCP inhibitor (PRCP-7414, referred to as PRCPi in text) was from Calbiochem (catalog number 504044).
7
Pharmacological Inhibition Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
RPMI 1640, penicillin/streptomycin solution, fetal bovine serum (FBS) were obtained from Invitrogen (Camarillo, CA, USA). Ketoconazole (KTZ) and sunitinib were purchased from Selleckchem (Houston, TX). U0126 and SB203580 were procured from Promega (Madison, WI, USA) and SP600125 from Sigma (Sigma-Aldrich, St. Louis, MO, USA). Unless otherwise indicated, all other drugs were purchased from Sigma (St. Louis, MO, USA)11 (link),12 (link).
8
Establishing Sunitinib-Resistant Renal Cancer Cell Lines
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Human immortalized normal kidney cell HK2, human papillary renal carcinoma cell (RCC) line ACHN, and clear RCC line 786-O were purchased from Cell Bank of the Chinese Academy of Sciences (Shanghai, China). Cells were cultured in Dulbecco’s modified Eagle’s medium (DMEM) (Gaithersburg, MD, United States) containing 10% fetal bovine serum (FBS) (Gaithersburg, MD, United States).
The parental ACHN and 786-O cells were treated with sunitinib Malate (Selleck Chemicals, Houston, TX, United States) at an initial dose of 0.25 μM for 72 h. Subsequently, the concentrations of sunitinib increased by 0.25 μM every 72 h. The survival clones were subsequently passaged until the IC50 values reached 18 μM in the 10th month. The stable sunitinib-resistant sublines were maintained with the medium supplemented by 0.02 μM sunitinib.
The parental ACHN and 786-O cells were treated with sunitinib Malate (Selleck Chemicals, Houston, TX, United States) at an initial dose of 0.25 μM for 72 h. Subsequently, the concentrations of sunitinib increased by 0.25 μM every 72 h. The survival clones were subsequently passaged until the IC50 values reached 18 μM in the 10th month. The stable sunitinib-resistant sublines were maintained with the medium supplemented by 0.02 μM sunitinib.
9
Breast Cancer Cell Colony Formation
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
For the colony formation assays, BT474 TRZ_R and HCC1954 breast cancer cells were plated into 6-well plates (2 × 103 cells/well) and incubated at 37 °C, O/N. The next day, the cells were treated with 2 μM ponatinib or sunitinib (Selleck Chemicals) and then incubated for an additional 10 days. Cell colonies were fixed with 10% ethanol, stained with 0.01% crystal violet, and observed using a CK40 inverted microscope (Olympus, Tokyo, Japan).
10
Isolation and Culture of Primary ATC Cells
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!