MHCC97H (97H), a high-metastatic human HCC cell line, was established in Liver Cancer Institute of Zhongshan Hospital [15 (link)]. MHCC97H cells from the 12th to the 15th passage were used in our experiments. Hep3B, a low metastatic potential HCC cell line was purchased from the America Type Culture Collection(ATCC, HB 8064™). The oxaliplatin-resistant HCC cell lines MHCC97H–OXA and Hep3B–OXA selected at a 25umol/L concentration of oxaliplatin were successfully established from MHCC97H and Hep3B, by exposing cells to gradually increasing oxaliplatin (Sigma, St. Louis, MO, USA) from 2 umol/L to 25umol/L in our laboratory [14 (link)]. The IC50 value of surviving HCC cells treated with oxaliplatin was about 10-fold as high as that of their parental cells (Additional file 1 : Figure S1).MHCC97H and MHCC97H–OXA were cultured in Dulbecco’s Modified Eagle’s Medium (DMEM, Gibco Invitrogen, Carlsbad, CA, USA) supplemented with 10% fetal bovine serum (FBS, Life Technologies/Gibco). Hep3B and Hep3B–OXA cells were cultured in minimum essential medium (MEM) supplemented with 10% fetal bovine serum. Cells(1 × 106) were seeded into 25 cm culture flask for 72 h per passage.
Oxaliplatin
Manufactured by Thermo Fisher Scientific
Sourced in United States
Oxaliplatin is a platinum-based chemotherapy medication used in the treatment of various types of cancer. It functions as a cytotoxic agent, disrupting the DNA structure and inhibiting cell division in rapidly dividing cancer cells.
Automatically generated - may contain errors
Lab products found in correlation
Oxaliplatin, by Merck Group (2 mentions)
Dmem culture medium, by Thermo Fisher Scientific (2 mentions)
Hep3b, by Merck Group (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (1 mentions)
Sorafenib, by Thermo Fisher Scientific (1 mentions)
Sorafenib, by Bayer (1 mentions)
Oxaliplatin, by Selleck Chemicals (1 mentions)
Annexin 5 pi staining kit, by Thermo Fisher Scientific (1 mentions)
Penicillin, by Thermo Fisher Scientific (1 mentions)
Rpmi 1640 medium, by Thermo Fisher Scientific (1 mentions)
Oxaliplatin, by Sanofi (1 mentions)
6 protocols using oxaliplatin
1
Establishment of Oxaliplatin-resistant HCC Cell Lines
2
Dissolving Oxaliplatin and Sorafenib
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Oxaliplatin was purchased from Sigma-Aldrich; Merck KGaA (Darmstadt Germany). Sorafenib was synthesized at Bayer (Newbury, UK). Oxaliplatin and Sorafenib were dissolved in Dulbecco's modified Eagle's medium (DMEM) containing 0.1% dimethylsulfoxide (Invitrogen; Thermo Fisher Scientific, Inc., Waltham, MA, USA).
3
Oxaliplatin-Induced Apoptosis in Pancreatic Cancer Cells
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Panc-1 cells were treated with 50 μM oxaliplatin (S1224, Selleck, Shanghai, China) for 48 h. MIAPaCa-2 cells were treated with 30 μM oxaliplatin for 48 h. Cell apoptosis was detected by using an Annexin V-PI staining kit (BMS500FI, Invitrogen, USA). Briefly, the cells were harvested and resuspended in Annexin V binding buffer with FITC-conjugated Annexin V and PI dye for 15 min. Then, the cells were analyzed using flow cytometer within 1 h. The experiment was repeated three times.
4
Establishing Oxaliplatin-Resistant LoVo Cells
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LoVo cells or OR-LoVo cells were cultivated in RPMI 1640 medium (GibcoTM, Invitrogen Corporation, France) containing 10% FBS (characterized fetal bovine serum; HyClone, Logan, UT, USA), and 1% penicillin (Invitrogen Corp., Carlsbad, CA, USA) and were then incubated in humidified air with 5% CO2 at 37 °C.
Oxaliplatin-resistant LoVo cells (OR-LoVo cells) were established from LoVo cells following the protocol in our previous study [10 (link)]. In this study, the drug-resistant colorectal cancer cell line was established by exposing a LoVo cell line to doses of 0–25 μM Oxaliplatin (OXA), which was changed from IV-injected liquid medicine (Sanofi, Paris, France) obtained from Dr. Ming-Cheng Chen [10 (link)] to the purified drug (Sigma, St. Louis, MO, USA), in a dose-dependent manner. The IC50 (50% cell death) of OXA in the LoVo cell line was found to be 15 μM, and this concentration was used to induce 90% cell death. The surviving cells were recovered to 80% in a culture plate and then passaged in the same OXA concentration to increase the OXA dose. In this study, we selected a cell population whose survival was under 3-fold the OXA IC50 concentration (45 μM) and identified this population as the OXA-resistant LoVo cell line.
Oxaliplatin-resistant LoVo cells (OR-LoVo cells) were established from LoVo cells following the protocol in our previous study [10 (link)]. In this study, the drug-resistant colorectal cancer cell line was established by exposing a LoVo cell line to doses of 0–25 μM Oxaliplatin (OXA), which was changed from IV-injected liquid medicine (Sanofi, Paris, France) obtained from Dr. Ming-Cheng Chen [10 (link)] to the purified drug (Sigma, St. Louis, MO, USA), in a dose-dependent manner. The IC50 (50% cell death) of OXA in the LoVo cell line was found to be 15 μM, and this concentration was used to induce 90% cell death. The surviving cells were recovered to 80% in a culture plate and then passaged in the same OXA concentration to increase the OXA dose. In this study, we selected a cell population whose survival was under 3-fold the OXA IC50 concentration (45 μM) and identified this population as the OXA-resistant LoVo cell line.
5
Oxaliplatin-Induced Mechanical Hyperalgesia
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To induce mechanical hyperalgesia, oxaliplatin (SAGENT pharmaceuticals, IL) was administered intraperitoneally at 3 mg/Kg for mice and 2 mg/Kg for rats for five consecutive days at a total dose of 15 mg/Kg and 10 mg/Kg, respectively. For in vitro culture experiment, oxaliplatin was diluted with DMEM culture medium (Gibco) at indicated concentrations.
6
Oxaliplatin-Induced Mechanical Hyperalgesia
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