| 名稱 | Doxycycline (hyclate) |
| 描述 | Doxycycline hyclate (WC2031) belongs to the tetracycline class of antibiotics and is a broad-spectrum metalloproteinase (MMP) inhibitor with oral activity. Doxycycline hyclate has antibacterial activity and antitumor activity. |
| 細(xì)胞實(shí)驗(yàn) | Doxycycline is diluted in culture medium at a concentration of 10 μg/mL. All in vitro treatments are performed in SMC cultures at 90% confluence, when ECM synthesis in SMC starts to be evident. Doxycycline (20 nM) is diluted in culture medium at a concentration of 10 μg/mL (20 nM), at which no toxicity or variation in primary cultured SMC proliferation has been reported, as well as in other cell lines and the incubation time is 48 h. SMC-C and SMC-Ch are seeded at equal cell density in 6-well plates and, when confluence reaches 90%, 1 mL culture medium is added to each well containing 3.7×104?Bq L-[5-3H]-proline (9.62×1011 Bq/mmol). After 48?h incubation, cells are lysed with 0.5 mL 0.5 mol/L NaOH for 1 h. The resulting solution is neutralized with an equal amount of 0.5 mol/L HCl, and 50 μL are used to measure total proteins with the Bradford method. One volume of 10% TCA is added to the remaining 250 μL and centrifuged at 13,000 g for 15?min at 4°C. The resulting precipitate is dissolved in 100 μL 0.2 mol/L NaOH, and then neutralized with 1 mol/L HCl. The solution is incubated with collagenase buffer (Tris-HCl, pH 7.6 20 mM, and CaCl2 250 mM final concentration) and 10 units of collagenase at 37°C overnight. Then, 150 μL 10% TCA are added and centrifuged at 13000 g for 15?min at 4°C. The resulting supernatant is added to 4 mL of scintillation fluid and the radioactivity is measured in a liquid scintillation counter LS 600 TA. |
| 激酶實(shí)驗(yàn) | Gelatin (0.1% (w/v) is added to standard LaemmLi acrylamide polymerization mixture. Tissue extract is mixed 1:2 with sample buffer [250 mM Tris-Cl pH 6.8, 10% (w/v) SDS, 20% (v/v) glycerol, 0.005% (w/v) bromphenol blue]. Serum is diluted 1:10 with electrophoresis buffer (2.5 mM Tris, 20 mM glycine, 0.005% SDS) and mixed 1:2 with sample buffer. Twenty μLs are loaded after 10-min incubation at room temperature without boiling. After electrophoresis at 90 V, the gels are soaked in 2.5% (w/v) Triton X-100, incubated 2 to 3 days at 37°C in gelatin digestion buffer [50 mM Tris-Cl, pH 8.0, 8 mM CaCl2, 10 mM ZnSO2, 0.02% (w/v) NaN3], stained in 0.05% Coomassie blue R-250 in acetic acid/methanol/water (1:4.5:4.5 by volume), destained in 10% acetic acid and 5% methanol, and scanned for lysis band intensity. The lysis band intensity is proportional to gelatinase activity and is quantified densitometrically by using One-Dimensional Scan software. The result, a number between 0.07 and 3.75, is normalized to the protein content by dividing the densitometry result with the relative optical density from the BCA protein assay kit result. The result is used for the analysis as the arbitrary unit. For the total MMP activity results of lysis bands of pro-MMP-9, active MMP-9, pro-MMP-2, and active MMP-2 are added. A protein size marker is used to determine the correct size. |
| 體外活性 | 方法：22 種腫瘤細(xì)胞用 Doxycycline hyclate 處理 48 h，使用 MTT 方法檢測(cè)細(xì)胞活力。
結(jié)果：肺癌細(xì)胞對(duì) Doxycycline 比其他大多數(shù)細(xì)胞系更敏感。NCI-H446 和 A549 細(xì)胞對(duì) Doxycycline 敏感，IC50分別為 1.70 μM 和 1.06 μM。[1]
方法：人胰腺癌細(xì)胞 PANC-1 用 Doxycycline hyclate (10-40 μg/mL) 處理 8-24 h，使用 Flow Cytometry 方法檢測(cè)細(xì)胞周期情況。
結(jié)果：Doxycycline 使細(xì)胞停滯在細(xì)胞周期的 G1-S 期。[2] |
| 體內(nèi)活性 | 方法：為檢測(cè)體內(nèi)抗腫瘤活性，將 Doxycycline hyclate (50 mg/kg) 腹腔注射給攜帶人乳腺癌腫瘤 4T1 的 BALB/c 小鼠，每天一次，持續(xù)十五天。
結(jié)果：Doxycycline 延遲腫瘤生長(zhǎng)并降低腫瘤相關(guān)巨噬細(xì)胞和血管的數(shù)量。[3]
方法：為檢測(cè)體內(nèi)抗菌活性，將 Doxycycline hyclate (40 mg/kg) 腹腔注射給暴露于 B. pseudomallei 1026b 的 BALB/c 小鼠，每天兩次，持續(xù)兩周。
結(jié)果：用 Doxycycline 治療的小鼠 100% 存活，并且從大多數(shù)存活小鼠的肺或脾臟中沒有擴(kuò)增出 B. pseudomallei 的 DNA。[4] |
| 存儲(chǔ)條件 | Powder: -20°C for 3 years | In solvent: -80°C for 1 year | Shipping with blue ice. |
| 溶解度 | H2O : 13.89 mg/mL, Sonication is recommended.
DMSO : 55 mg/mL (107.23 mM), Sonication is recommended.
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| 關(guān)鍵字 | U343 | anti-proliferative activity | SVG | MCF 12A | WC-2031 | antibacterial activity | Matrix metalloproteinases | 293T | LNT-229 | hyclate | inhibit | Doxycycline (hyclate) | WC 2031 | Antibiotic | MMP | Doxycycline Hydrochloride | Inhibitor | Bacterial | G55 | Doxycycline |
| 相關(guān)產(chǎn)品 | Crystal Violet | Dehydroacetic acid sodium | Doxycycline | Methyl anthranilate | Glucosamine | Neomycin sulfate | Metronidazole | Dimethyl sulfoxide | EDTA copper(II) disodium salt | Ampicillin sodium | Sulfamethoxazole sodium | Kanamycin sulfate |
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