Mitomycin C (Ametycine) is a DNA cross-linking agent and induces DNA damaging. Mitomycin C is an antitumor agent and antibiotic that shows extraordinary ability to inhibit DNA synthesis. Mitomycin C is an ADC Cytotoxin and induces apoptosis^[1][2][3].

The HCT116 (p53^-/-) cells are minimally sensitive to either Mitomycin C (Ametycine) or TRAIL alone. However, surprisingly, combination treatment with MMC and TRAIL decreases cell viability significantly. Although Mitomycin C and TRAIL alone are moderately effective, Mitomycin C substantially enhances the effect of TRAIL on suppression of the cell proliferation. Mitomycin C and TRAIL treatment alone induces 9.5% and 35.0% apoptosis, respectively. However, combination treatment with Mitomycin C and TRAIL enhances apoptosis to 66.6%^[1].
Mitomycin C is a cytotoxic chemotherapeutic agent that causes DNA damage in the form of DNA cross-links as well as a variety of DNA monoadducts and is known to induce p53^[2].

上海金畔生物科技有限公司 has not independently confirmed the accuracy of these methods. They are for reference only.

Mice bearing xenografted HCT116 (p53^-/-) colon tumors and HT-29 colon tumors are treated with Mitomycin C (Ametycine; i.p., 1 mg/kg) and TRAIL (i.v., 100 μg) every other day. Animals are treated with 10 consecutive cycles of the combination therapy regimen. The combination therapy suppresses tumor growth significantly and does not impact the weight of the mice, indicating that the therapeutic combination of Mitomycin C and TRAIL is well-tolerated and has anti-tumor activity in vivo^[1].
Intravesical Mitomycin C instillations has an effect on body weight. After 3 consecutive weekly instillations of 1 mg/mL Mitomycin C there is almost no weight gain, whereas rats in the other 3 groups has a statistically significant weight gain compared with MMC treated rats^[3].

上海金畔生物科技有限公司 has not independently confirmed the accuracy of these methods. They are for reference only.

334.33

C₁₅H₁₈N₄O₅

50-07-7

丝裂霉素 C；丝裂霉素；密吐霉素；自力霉素；嘧吡霉素；突变霉素

Room temperature in continental US; may vary elsewhere.

4°C, protect from light

*In solvent : -80°C, 6 months; -20°C, 1 month (protect from light)

In Vitro: 

DMSO : 50 mg/mL (149.55 mM; Need ultrasonic)

H₂O : < 0.1 mg/mL (ultrasonic;warming;heat to 60°C) (insoluble)

*

请根据产品在不同溶剂中的溶解度选择合适的溶剂配制储备液；一旦配成溶液，请分装保存，避免反复冻融造成的产品失效。
储备液的保存方式和期限：-80°C, 6 months; -20°C, 1 month (protect from light)。-80°C 储存时，请在 6 个月内使用，-20°C 储存时，请在 1 个月内使用。

In Vivo:

请根据您的实验动物和给药方式选择适当的溶解方案。以下溶解方案都请先按照 In Vitro 方式配制澄清的储备液，再依次添加助溶剂：

——为保证实验结果的可靠性，澄清的储备液可以根据储存条件，适当保存；体内实验的工作液，建议您现用现配，当天使用； 以下溶剂前显示的百
分比是指该溶剂在您配制终溶液中的体积占比；如在配制过程中出现沉淀、析出现象，可以通过加热和/或超声的方式助溶

• 1.

  请依序添加每种溶剂： 10% DMSO    40% PEG300    5% Tween-80    45% saline

  Solubility: ≥ 2.08 mg/mL (6.22 mM); Clear solution

  此方案可获得 ≥ 2.08 mg/mL (6.22 mM，饱和度未知) 的澄清溶液。

  以 1 mL 工作液为例，取 100 μL 20.8 mg/mL 的澄清 DMSO 储备液加到 400 μL PEG300 中，混合均匀；向上述体系中加入50 μL Tween-80，混合均匀；然后继续加入 450 μL生理盐水定容至 1 mL。

  将 0.9 g 氯化钠，完全溶解于 100 mL ddH₂O 中，得到澄清透明的生理盐水溶液

• 2.

  请依序添加每种溶剂： 10% DMSO    90% (20% SBE-β-CD in saline)

  Solubility: ≥ 2.08 mg/mL (6.22 mM); Clear solution

  此方案可获得 ≥ 2.08 mg/mL (6.22 mM，饱和度未知) 的澄清溶液。

  以 1 mL 工作液为例，取 100 μL 20.8 mg/mL 的澄清 DMSO 储备液加到 900 μL 20% 的 SBE-β-CD 生理盐水水溶液中，混合均匀。

  将 2 g 磺丁基醚 β-环糊精加入 5 mL 生理盐水中，再用生理盐水定容至 10 mL，完全溶解，澄清透明

• [1]. Cheng H, et al. Mitomycin C potentiates TRAIL-induced apoptosis through p53-independent upregulation of death receptors: Evidence for the role of c-Jun N-terminal kinase activation. Cell Cycle. 2012 Sep 1;11(17): 3312-23.

  [2]. Abbas T, et al. Differential activation of p53 by the various adducts of mitomycin C. J Biol Chem. 2002 Oct 25;277(43):40513-9.

  [3]. Michielsen D, et al. Mitomycin C: functional bladder damage in rats after repeat intravesical instillations. J Urol. 2005 Jun;173(6):2166-70.

Colon adenocarcinoma HCT116 and HT-29 human colon cancer cells are used. The CellTiter-Glo Luminescent Cell Viability Assay is used to measure cell viability, which use a unique, stable form of luciferase to measure ATP as an indicator of viable cells, and the luminescent signal produced is proportional to the number of viable cells present in culture. Cells are pretreated with Mitomycin C (5 μM) for 12 h or 24 h, and then exposed to different concentrations of TRAIL for 12 h. An equal volume (100 μL) of CellTiter-Glo^TM reagent is added and the solution is mixed gently for 2 min on an orbital shaker. Mixture is incubated at room temperature for 10 min to allow luminescent signal to stabilize, and then imaging is performed using the Xenogen IVIS system to quantify the cell viability^[1].

上海金畔生物科技有限公司 has not independently confirmed the accuracy of these methods. They are for reference only.

Mice^[1]
Four- to 6-wk-old NCr nude mice are treated with Mitomycin C (1 mg/kg) by intraperitoneal injection for 24 h, followed by one intravenous dose of purified rhTRAIL (100 μg). As a negative control, a subset of the mice are injected (i.p. and i.v.) with saline (vehicle) at the same frequency of treatment. Animals are treated for 3 consecutive weeks. The tumor size is monitored every week using caliper measurements of the tumor volume.
Rats^[3]
Young adult female Wistar rats at age 13 weeks with a median weight of 217 g (range 187 to 255) are randomized into 4 groups of 10 each, namely a normal group with no instillations, an NaCl 0.9% or placebo group that received instillations with the solvent of the chemotherapeutic agent, Mitomycin C (1 mg/mL) group.

上海金畔生物科技有限公司 has not independently confirmed the accuracy of these methods. They are for reference only.

• [1]. Cheng H, et al. Mitomycin C potentiates TRAIL-induced apoptosis through p53-independent upregulation of death receptors: Evidence for the role of c-Jun N-terminal kinase activation. Cell Cycle. 2012 Sep 1;11(17): 3312-23.

  [2]. Abbas T, et al. Differential activation of p53 by the various adducts of mitomycin C. J Biol Chem. 2002 Oct 25;277(43):40513-9.

  [3]. Michielsen D, et al. Mitomycin C: functional bladder damage in rats after repeat intravesical instillations. J Urol. 2005 Jun;173(6):2166-70.