Method of estimating antitumor effect of histone deacetylase inhibitor.
A technique for sirtuins and inhibitors, applied in the field of predicting the anti-tumor effect of histone sirtuin inhibitors
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Embodiment 1
[0074] Evaluation of compound A sensitive tumors and compound A resistant tumors
[0075] (1) Reagent preparation
[0076]Weigh the necessary amount of FK228, add a solvent (10% HCO-60 / salt water), and perform ultrasonic dissolution. Before the start of the experiment, the positive control substance, paclitaxel, was dissolved in Cremophor EL / ethanol (1:1) solution and stored under refrigeration. When used, it was diluted by adding 9 times the amount of normal saline to make 2.4 mg / mL (solvent composition: 5% Cremophor EL-5% ethanol-90% saline).
[0077] (2) Experimental animals
[0078] In the anti-tumor test of the drug, BALB / cANnNCrJ-nu / nu mice (male, 6 weeks old) were purchased from Charles River of Japan and used for the test after being domesticated for one week or more. The mice are raised in an SPF environment and freely take in water and food.
[0079] (3) Experimental tumor
[0080] Divide 2~3×10 7 Human cultured kidney cancer cell line 1 (ACHN: obtained from ATCC) and hu...
Embodiment 2
[0091] Evaluation of compound A sensitive tumors and compound A resistant tumors
[0092]
[0093] (1) Experimental materials
[0094] Pharmaceutical Compound A (FK228)
[0095] Dosage: 3.2mg / kg
[0096] Dosing capacity: 10mL / kg
[0097] Solvent: 10% HCO-60 / saline solution
[0098] Dosage form: solution (when used)
[0099] Tumor cell human prostate cancer PC-3 (tumor section 3mm×3mm×3mm / mouse inoculation site s.c.)
[0100] Human gastric cancer SC-6; obtained by the Central Laboratory Animal Research Institute
[0101] (Tumor section 3mm×3mm×3mm / mouse inoculation site s.c.)
[0102] Human renal carcinoma ACHN (Tumor section 3mm×3mm×3mm / mouse transplantation site s.c.)
[0103] Human renal carcinoma A498; Obtained from ATCC (Tumor section 3mm×3mm×3mm / mouse inoculation site s.c.)
[0104] Subordinate animal male BALB c / nu / nu
[0105] (2) Sequence and result
[0106] According to the method shown in Example 1, a 3mm square tumor section (human prostate cancer PC-3, human gastric ca...
Embodiment 3
[0108] Analyze the gene expression patterns of compound A-sensitive tumors and compound A-resistant tumors by gene chip
[0109] In Example 2, gene expression patterns in tumors that can be confirmed as sensitive or resistant were studied.
[0110] (1) Experimental materials
[0111] Tumor cell human prostate cancer PC-3 (tumor section 3mm×3mm×3mm / mouse inoculation site s.c.)
[0112] Human gastric cancer SC-6; obtained from the Central Laboratory of Experimental Animals (Tumor Section 3mm×3mm×3mm / mouse inoculation site s.c.)
[0113] Human renal cancer ACHN (Tumor section 3mm×3mm×3mm / mouse inoculation site s.c.)
[0114] Human renal carcinoma A498; Obtained from ATCC (Tumor section 3mm×3mm×3mm / mouse inoculation site s.c.)
[0115] Subordinate animal male BALB c / nu / nu
[0116] RNA extraction Rneasy Mini Kit (50) (Qiagen)
[0117] RNase, DNase free water (Life Technologies)
[0118] DNA Synthesis Superscript Choice System (Life Technologies)
[0119] Etchachinmate(Nippon gene)
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