Cisplatin

Manufactured by Meilun

Sourced in China

Cisplatin is a platinum-based inorganic compound used as a chemotherapeutic agent in the treatment of various types of cancer. It functions as a cytotoxic drug, interfering with DNA replication and transcription, ultimately leading to cell death.

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Lab products found in correlation

Methyl thiazolyl tetrazolium (mtt), by Merck Group (2 mentions) Dimethyl sulfoxide (dmso), by Merck Group (2 mentions) Microplate reader, by Thermo Fisher Scientific (2 mentions) Anti lc3a b antibody, by Cell Signaling Technology (1 mentions) Anti rabbit igg, by ZSGB-BIO (1 mentions) Anti mouse igg, by ZSGB-BIO (1 mentions) Anti nqo1, by Abcam (1 mentions) Antibodies against β actin, by ZSGB-BIO (1 mentions) Osthole, by Meilun (1 mentions) Dicoumarol, by MedChemExpress (1 mentions) 2 7 dichloro uorescin diacetate dcfh da, by Merck Group (1 mentions) 3 methyladenine, by MedChemExpress (1 mentions) Anti caspase 3, by Abcam (1 mentions) Anti gsdme, by Abcam (1 mentions) Anti parp 1, by Abcam (1 mentions) Z devd fmk, by MedChemExpress (1 mentions) Anti cleaved caspase 3, by Abcam (1 mentions) Fitc annexin 5 apoptosis detection kit, by BD (1 mentions) Z vad fmk, by MedChemExpress (1 mentions) N acetyl l cysteine nac, by Beyotime (1 mentions)

14 protocols using cisplatin

Regulation of HDAC2 Stability by USP43 in Ovarian Cancer

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HEY and OV-90 cells were purchased from Guangzhou Cellcook Biotech Co., Ltd. HEY cells were cultured in RPMI-1640 medium (Solarbio Science), which included 10% fetal bovine serum (FBS) (Tianhang Biotechnology). OV-90 cells were cultured in DMEM/F12 medium (Biosharp Life Science), which supplemented with 15% FBS. All cells were cultured in in an incubator at 37 °C with 5% CO₂.
Two USP43 shRNA vectors and USP43 overexpression vector were designed and cloned into transfer plasmids. Lentiviral particles were produced using 293T cells and harvested to infect HEY and OV-90 cells. Puromycin was added into the medium to obtain stably infected cells. For cisplatin sensitivity experiments, cells were treated with the indicated concentrations of cisplatin (meilunbio) for 48 h. To investigate the effect of USP43 on the stability of HDAC2, stable transfected cells were treated with 10 µM cycloheximide (Med Chem Express) for 0, 2, 4 and 6 h, respectively. For the detection of HDAC2 ubiquitination levels, stable transfected cells were pretreated with 20 µM MG132 (Aladdin) for 8 h.
HDAC2 knockdown in HEY cells were carried out using HDAC2 siRNA (si-HDAC2: 5’-GGUCAAUAAGACCAGAUAATT-3’) via Lipofectamine 3000 (Invitrogen), and si-NC as the negative control.

Pei L., Zhao F, & Zhang Y. (2023). USP43 impairs cisplatin sensitivity in epithelial ovarian cancer through HDAC2-dependent regulation of Wnt/β-catenin signaling pathway. Apoptosis, 29(1-2), 210-228.

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Cell Proliferation and Cisplatin Sensitivity Assays

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To detect cell proliferation ability, cells (1 × 10⁴/well) were seeded in 96-well plates and transfected with OE-TBX1 plasmids, miR-6727-5p inhibitor, TBX1 siRNA or their negative control for 0, 24, 48, 72 and 96 hours. To test the cisplatin sensitivity of CC cells, cells (1 × 10⁴/well) were seeded in 96-well plates and transfected with OE-TBX1 plasmids or empty vectors for 24 hours, then treated with different concentrations of cisplatin (0, 2, 4, 8, 16, 32 and 64 μM; MeilunBio, China) for additional 24 hours. After the treatment, cells were incubated with CCK-8 regents (KeyGEN, China) for 2 hours, and the optical density (OD) at 450 nm was measured with a microplate reader (BIOTEK, USA).

Liu H., Song M., Sun X., Zhang X., Miao H, & Wang Y. (2021). T-box transcription factor TBX1, targeted by microRNA-6727-5p, inhibits cell growth and enhances cisplatin chemosensitivity of cervical cancer cells through AKT and MAPK pathways. Bioengineered, 12(1), 565-577.

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Cytotoxicity Assay of Cisplatin and 5-FU

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The resuspended circ-BMI1/mock transfected cells were cultured in a 96-well plate in complete medium with a density of 5000/well cells and a volume of 50 μL. Added different concentrations of cisplatin (Meilun Biotechnology, China) and 5-fluorouracil (Meilun Biotechnology, China) prepared with complete medium to each well, totaling 50 μL. The cisplatin concentrations were 25 μmol/L, 50 μmol/L, 100 μmol/L, 150 μmol/L, 200 μmol/L, and the concentrations of 5-fluorouracil were 2.5 μmol/L, 10 μmol/L, 40 μmol/L, 80 μmol/L, 160 μmol/L, respectively. 10 µL of CCK-8 was added the next day. After 3 h of incubation, the OD value at 450 nm was measured by a microplate reader to calculate the growth inhibition rate (IR).

Zhao Q., Zhu X., Ke J.M., Su X.Y., Yi J., Wu D.L., Lin J, & Deng Z.Q. (2021). Circular RNA BMI1 Serves as a Potential Target for Diagnosis and Treatment in Esophageal Cancer. Technology in Cancer Research & Treatment, 20, 15330338211033075.

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Cisplatin Cytotoxicity Assay in 96-Well Plates

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Cells were seeded in 96-well plates at a density of 2×10 3 per well and cultured to 80% confluence in a 37˚C, 5% CO2 incubator. Cells were then exposed to cisplatin (Meilun, Dalian, China) of an ascending concentration range (0.75, 1.5, 3, 6, 12, 20, 40 μM) for 48h and 72h respectively, with five replicates for each testing point including untreated controls (0 μM cisplatin for 48h and 72h) and blank wells. Thereafter, cells were incubated with 8 μl MTT (0.2 mg/ml) (Sigma-Aldrich, St. Louis, MO, USA) for 4h followed by incubation with 200 μl DMSO (Sigma-Aldrich, St. Louis, MO, USA) to dissolve the crystals before reading the absorbance at 490 nm in a microplate reader (BIOTEK, Vermont, USA). The results are expressed as inhibitory rates and cisplatin IC 50 values (the concentrations resulting in 50% inhibition).

Zhao W.J., Deng B.Y., Wang X.M., Miao Y, & Wang J.N. (2015). XIAP associated factor 1 (XAF1) represses expression of X-linked inhibitor of apoptosis protein (XIAP) and regulates invasion, cell cycle, apoptosis, and cisplatin sensitivity of ovarian carcinoma cells. Asian Pacific journal of cancer prevention : APJCP, 16(6).

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Osthole and Cisplatin Co-treatment Induces Apoptosis

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Osthole and cisplatin were purchased from Meilunbio (purity≥98%, Dalian, China). Dicoumarol (DIC), Z-VAD-FMK, Z-DEVD-FMK and 3-Methyladenine (3-MA) were purchased from MedChem Express (Princeton, NJ, USA). N-Acetyl-L-cysteine (NAC) and JC-1 were purchased from Beyotime (Shanghai, China). Dimethyl sulfoxide (DMSO) and 2′, 7′-Dichlor-o uorescin diacetate (DCFH-DA) were purchased from Sigma-Aldrich (St. Louis, MO, USA). Anti-rabbit IgG, anti-mouse IgG and antibodies against β-actin were purchased from ZSGB-BIO (Beijing, China). Anti-LC3A/B antibody was purchased from Cell Signaling Technology (Danvers, MA, USA). Anti-NQO1, anti-caspase-3, anti-cleaved caspase-3, anti-PARP-1 and anti-GSDME antibodies were purchased from Abcam (Cambridge, MA, USA). FITC Annexin V Apoptosis Detection Kit (BD Pharmingen, CA, USA)

Huangfu M., Wang J., Yu D., Qin J., Guan X., Zhou L., Dou T., Liu Y., Wang L., Li X., Fu M., & Xu C. (2020). Osthole Induces Apoptosis, Secondary Necrosis and Mitophagy Via NQO1-Mediated ROS Overproduction in HeLa Cells.

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Generation of Brain-Metastatic Lung Cancer Cell Line

Cited 2 times

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The human lung cancer cell line PC9 was obtained from the Chinese Academy of Medical Sciences (Beijing, China). The BM derivative cells (PC9‐BrM1, PC9‐BrM2, and PC9‐BrM3) were derived from parental cell line PC9 by implanting tumor cells in immunodeficient mice via left ventricular injection; the metastatic cells were extracted from the harvested brain metastases as described in previous work.⁹ (link) PC9‐BrM3 was created with two more rounds of injection‐extraction‐expansion cycling; it has been shown to exhibit enriched brain metastasis activity.²⁴ (link), ²⁵ (link) These cells were cultured in Roswell Park Memorial Institute medium‐1640 (RPMI1640) with the supplementation of 10% fetal bovine serum (FBS), 100 U/mL penicillin and streptomycin (all those agents were obtained from Gibco, Invitrogen, Inc, USA) and a humidified environment with 5% CO2 at 37°C was used to maintain.
Cisplatin (MB1055), Carboplatin (MB1297), Ferrostatin‐1 (Fer‐1, MB4718), (1S, 3R)‐RSL3 (RSL3, MB4723), TGF‐β inhibitor SB431542 (MB5459), Wnt/β‐catenin inhibitor I WR‐1‐endo (MB4050), p38 MAPK inhibitor SB203580 (MB5062), and ERK inhibitor SCH772984 (MB3432) were purchased from Meilunbio (China). Wnt agonist 1 (B6059), Cycloheximide (CHX, A8244) was obtained from ApexBio (USA).

Liu W., Zhou Y., Duan W., Song J., Wei S., Xia S., Wang Y., Du X., Li E., Ren C., Wang W., Zhan Q, & Wang Q. (2021). Glutathione peroxidase 4‐dependent glutathione high‐consumption drives acquired platinum chemoresistance in lung cancer‐derived brain metastasis. Clinical and Translational Medicine, 11(9), e517.

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Evaluating Anti-Cancer Treatments in Xenograft Model

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H1975 cells (1 × 10⁶/ml) were inoculated subcutaneously into the right thigh of male 4 to 6 weeks old of Nu/Nu mice (Charles River). When the average tumor volume reached 200 mm³, the mice were randomized into 5 groups to receive the following treatments: (a) vehicle (methycellulose/Tween 80, per os) (Dalian Meilun Biotech); (b) CQ (50 mg/kg, intraperitoneal injection); (c) afatinib (20 mg/kg/day, per os); (d) afatinib (20 mg/kg/day, per os) + CQ (50 mg/kg, intraperitoneal injection) (e) cisplatin (10 mg/kg, every other day, intraperitoneal injection) (Dalian MeilunBiotech). Tumors were measured every 3 days, and tumor volumes were determined from caliper measurements of tumor length (L) and width (W) according to the formula V = (L × W²)/2. All animal procedures were performed in accordance with institutional guidelines and with approval from Fudan University Committee on Animal Care.

Hu X., Shi S., Wang H., Yu X., Wang Q., Jiang S., Ju D., Ye L, & Feng M. (2017). Blocking autophagy improves the anti-tumor activity of afatinib in lung adenocarcinoma with activating EGFR mutations in vitro and in vivo. Scientific Reports, 7, 4559.

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Cisplatin and Simotang Oral Liquid Protocol

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Cisplatin (DDP, a commonly used drug in combination chemotherapy) was purchased from Dalian Meilun Biotechnology Co., Ltd. (China) and Simotang Oral Liquid (SMT) was purchased from Hunan Hansen Pharmaceutical Co., Ltd. (China). Antibiotics were purchased from Sigma-Aldrich (China) Trading Co., Ltd. Other chemicals and reagents used in this study were of analytical grade.

Deng L., Zhou X., Lan Z., Tang K., Zhu X., Mo X., Zhao Z., Zhao Z, & Wu M. (2022). Simotang Alleviates the Gastrointestinal Side Effects of Chemotherapy by Altering Gut Microbiota. Journal of Microbiology and Biotechnology, 32(4), 405-418.

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Investigating SGK1 Role in Cell Proliferation

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To study the function of SGK1 on cell proliferation, the cells at the density of 5×10³ cells /well were seeded in a 96-well plate of y13B (LIU YI, Beijing, China; AF2006, Affinity). Vector construction and cell transfection or cells were treated with cisplatin (MB1055, meilunbio, Dalian) at 0, 0.25, 0.5, 1, 1.5, 2, 3, 4, and 6 μM after cell transfection. After further growth for different durations, 10 μL CCK-8 solution (96992, Sigma, USA) per well was added. After treatment at 37 °C for 1 h, the absorbance value at 450 nm was detected by a microplate reader (800Ts, BioTek, USA).

Zhang J., Lv W., Liu Y., Fu W., Chen B., Ma Q., Gao X, & Cui X. (2021). Knockdown of Serum- and Glucocorticoid-Regulated Kinase 1 Enhances Cisplatin Sensitivity of Gastric Cancer Through Suppressing the Nuclear Factor Kappa-B Signaling Pathway. Balkan Medical Journal, 38(6), 331-340.

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Cytotoxicity Assays of Anticancer Drugs

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Nelfinavir mesylate (CAS:159989-65-8), Adriamycin (CAS:25316-40-9), colchicine (CAS:64-86-8), paclitaxel (CAS:33069-62-4), imatinib mesylate (CAS:220127-57-1), and cisplatin (CAS:15663-27-1)
were purchased from Dalian Meilun Biotech Co., Ltd. (Dalian, China). All of the chemicals were dissolved in dimethyl sulfoxide and diluted with RPMI medium 1640 (Thermo Fisher Scientific, Waltham, MA). Cell Counting Kit-8 (CCK-8) kits were obtained from Bimake (Houston, TX). Annexin V Apoptosis Detection Kit FITC and SYTOX™ Red Dead Cell Stain were purchased from Invitrogen, Ltd. (Carlsbad, CA). Antibodies against P-gp were purchased from Cell Signaling Technology (Danvers, MA). Antibodies against ERK1/2, p-ERK1/2, Akt, p-Akt caspase-3, and cleaved caspase-3 were also obtained from Cell Signaling Technology. Bcl-2 and bax were purchased from the Proteintech Group (Wuhan, China).

Liu W., Meng Q., Sun Y., Wang C., Huo X., Liu Z., Sun P., Sun H., Ma X, & Liu K. (2018). Targeting P-Glycoprotein: Nelfinavir Reverses Adriamycin Resistance in K562/ADR Cells. Cellular physiology and biochemistry : international journal of experimental cellular physiology, biochemistry, and pharmacology, 51(4).

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