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3 protocols using fluvastatin sodium

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Statins Overcome Erlotinib Resistance in NSCLC

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Experiments were carried out with human lung adenocarcinoma cell lines A549 (ATCC CCL_185), Calu6 (ATCC HTB-56) and NCI-H1993 (ATCC CRL-5909). All cell lines were obtained from American Type Cell Culture Collection (ATCC) and cultured in DMEM growth medium (GIBCO #31966-21), supplemented with 10% heat-inactivated foetal bovine serum (FBS) and 1% antibiotics (penicillin, streptomycin). Cells were kept at 37 °C and 5% CO2 in the incubator and were passaged at 80–90% confluence every 2–3 days to maintain continuous logarithmic growth. All cell lines studied in this work were erlotinib resistant and EGFR wild type (Table 1). Cells were treated with pitavastatin calcium (SelleckChem, #S1759), fluvastatin sodium (SelleckChem, #S1909) and erlotinib hydrochloride (SelleckChem, #S1023).

Human NSCLC cell lines harbouring different genetic mutations examined in the study.

Cell lineEGFRK-RasMetp53Erlotinib
A549WildtypeMutatedWildtypeWildtypeResistant
Calu6WildtypeMutatedWildtypeMutatedResistant
H1993WildtypeWildtypeAmplifiedMutatedResistant
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2

Comprehensive Analytical Reagents Protocol

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Water (LC-MS Grade, Optima; 10509404), Acetonitrile (LC-MS Grade, Optima; 10001334), Methanol (LC-MS Grade, Optima, A456-212) and Formic acid (LC-MS Grade, Thermo Scientific Pierce; 13454279) were purchased from Fisher Chemicals. Human cell lysate (MS Compatible Human Protein Extract, Digest, V6951) and Trypsin (Sequence grade, V511X) were purchased from Promega. DL-Dithiothreitol (BioUltra, 43815), Iodoacetamide (BioUltra, I1149) Ammonium Bicarbonate (Eluent additive for LC-MS, 40867), Yeast Nitrogen Base Without Amino Acids (Y0626) and glass beads (acid washed, 425-600 µm, Sigma, G8772) were purchased from Sigma Aldrich. Urea (puriss. P.a., reag. Ph. Eur., 33247H) and Acetic Acid (Eluent additive for LC-MS, 49199) were purchased from Honeywell Research Chemicals. Rosuvastatin Calcium (S2169), Fluvastatin Sodium (S1909), Pyrimethamine (S2006), Pitavastatin Calcium (S1759), Pemetrexed Disodium Hydrate (S7785), Pravastatin Sodium (S3036), Clotrimazole (S1606), Miconazole Nitrate (S1956), Lovastatin (S2061), Ketoconazole (S1353), Atorvastatin (S2077), Methotrexate Disodium (S5097), Simvastatin (S1792), Uniconazole (S3660), Itraconazole (S2476) were purchased from Selleck chemicals and Pralatrexate (A4350) was purchased from APExBIO. Control samples for SARS-CoV-2 study were prepared from commercial Human Plasma (EDTA, Pooled Donor, Genetex GTX73265).
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3

Cholesterol Biosynthesis Pathway Modulators

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We purchased Pitavastatin Calcium (S1759), Fluvastatin sodium (S1909), ABT-199 (S0848), and CHIR-98014 (S2745) from Selleckchem. We purchased cholesterol (S5442), mevalonolactone (M4667), and coenzyme Q10 (C9538) from Sigma-Aldrich. We purchased YM-53601 (18113), farnesyl pyrophosphate ammonium salt (63250) and geranylgeranyl pyrophosphate ammonium salt (63330) from Cayman Chemical.
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