Colcemid
(Synonyms: 秋水仙碱,Demecolcine) 目录号 : GC40664Colcemid是一种细胞骨架抑制剂，可以在哺乳动物细胞中诱导G2/M期的有丝分裂停滞或在卵母细胞中诱导囊泡破裂（GVBD）期的减数分裂停滞。
Cas No.:477-30-5
Sample solution is provided at 25 µL, 10mM.
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Quality Control & SDS
- View current batch:
- Purity: >99.50%
- COA (Certificate Of Analysis)
- SDS (Safety Data Sheet)
- Datasheet
| Cell experiment [1]: | |
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Cell lines |
Syrian hamster embryo cells |
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Preparation Method |
Treatment of Syrian hamster embryo cells in culture with 0.01-0.1 µg/ml of colcemid for 48 h resulted in morphological and neoplastic transformation of the cells. |
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Reaction Conditions |
0.01-0.1 µg/ml of colcemid for 48 h |
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Applications |
Cell transformation was observed with doses which were non-cytotoxic and did not cause mitotic inhibition of the cells. Higher dose of colcemid (greater than 0.1 µg/ml) resulted in mitotic inhibition of the cells and a significant loss of colony forming ability, but no increase in the frequency of morphological transformation.A 14-fold increase in the number of aneuploid cells with a near diploid chromosome complement was found in cultures treated with 0.1 µg/ml colcemid and both chromosome loss and gain were induced. |
| Animal experiment [2]: | |
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Animal models |
Tumor bearing mice |
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Preparation Method |
Mice were injected intravenously with Colcemid |
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Dosage form |
5.82 µg/hr for 4h |
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Applications |
A mitotic linear accumulation was obtained by continuous colcemid infusion at 5.82 µg/hr. Low dose colcemid infusion (0.582 and 1.455 µg/hr) for 14 hours did not accumulated mitotic cells, but doses more than 5.82 µg/hr of colcemid blocked it completely, accumulating 25.5% of cells after a 20 hours infusion. |
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References: [1]. Tsutsui T, Maizumi H, et,al. Colcemid-induced neoplastic transformation and aneuploidy in Syrian hamster embryo cells. Carcinogenesis. 1984 Jan;5(1):89-93. doi: 10.1093/carcin/5.1.89. PMID: 6690091. |
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Colcemid is a cytoskeletal inhibitor that induces mitotic arrest in the G2/M phase or meiotic arrest in the vesicle rupture (GVBD) phase in mammalian cells or oocytes, respectively[1][2].Colchicine interferes with microtubule polymerization by tightly binding to tubulin dimers and prevents spindle microtubule formation by depolymerization [7].
Cell transformation was observed with doses which were non-cytotoxic and did not cause mitotic inhibition of the cells. Higher dose of colcemid (greater than 0.1 µg/ml) resulted in mitotic inhibition of the cells and a significant loss of colony forming ability, but no increase in the frequency of morphological transformation.A 14-fold increase in the number of aneuploid cells with a near diploid chromosome complement was found in cultures treated with 0.1 µg/ml colcemid and both chromosome loss and gain were induced[4]. Colcemid was more cytotoxic to cells in G2 + M than to G1 + S phase cells, and it slowed the progression of G1 cells to S. These effects of colcemid were much greater in aneuploid B16 melanoma cells than in pseudodiploid Chinese hamster ovary (CHO) cells[5]. Colcemid promotes UVC-induced apoptosis in Chinese hamster ovary cells (CHO.K1).Although colcemid did not affect the excision of UV-induced DNA damages such as photoproducts or cyclobutane pyrimidine dimers, colcemid accumulated the DNA breaks when it was added to cells following UV-irradiation[3].
A mitotic linear accumulation was obtained by continuous colcemid infusion at 5.82 µg/hr. Low dose colcemid infusion (0.582 and 1.455 µg/hr) for 14 hours did not accumulated mitotic cells, but doses more than 5.82 µg/hr of colcemid blocked it completely, accumulating 25.5% of cells after a 20 hours infusion[6].
Colcemid是一种细胞骨架抑制剂，可以在哺乳动物细胞或卵母细胞中诱导G2/M期的有丝分裂停滞或囊泡破裂（GVBD）期的减数分裂停滞[1][2]。而秋水仙碱则通过紧密结合到微管蛋白二聚体上干扰微管聚合，并通过去聚合作用阻止纺锤体微管形成[7]。
在非细胞毒性剂量下观察到了细胞转化，这些剂量不会引起细胞有丝分裂抑制。高剂量的科尔塞米德（大于0.1微克/毫升）导致细胞有丝分裂抑制和显著的菌落形成能力损失，但没有增加形态转化频率。用0.1微克/毫升科尔塞米德处理培养物中发现近二倍体染色体组合的非整倍体细胞数量增加了14倍，并诱导染色体缺失和增益。科尔塞米德对G2 + M期的细胞比对G1 + S期的更具有细胞性毒性，并减缓G1期向S期进展。这种影响在非整倍体B16黑色素瘤细胞中比伪二倍体中国仓鼠卵巢（CHO） 细胞更为明显。科尔塞米德促进UVC诱导的中国仓鼠卵巢（CHO.K1） 细胞凋亡。虽然科尔塞米德不影响紫外线诱导DNA损伤如光产物或环戊基嘧啶二聚物的切除，但当它在紫外线照射后加入细胞时，会积累DNA断裂。
通过持续注射5.82微克/小时的科尔赛米德，获得了有丝分裂线性积累。低剂量（0.582和1.455微克/小时）的科尔赛米德注射14个小时并没有积累有丝分裂细胞，但是超过5.82微克/小时的剂量完全阻止了它，并在20个小时的注射后积累了25.5% 的细胞。
References:
[1]. Tsuchida T, Yoshimura K, et,al. Colcemid-induced apoptosis of cultured human glioma: electron microscopic and confocal laser microscopic observation of cells sorted in different phases of cell cycle. Cytometry. 1998 Apr 1;31(4):295-9. doi: 10.1002/(sici)1097-0320(19980401)31:43.0.co;2-i. PMID: 9551605.
[2]. Ashley M Rozario, Sam DuwÉ, et,al.Ultra-Low Colcemid Doses Induce Microtubule Dysfunction as Revealed by Super-Resolution Microscopy bioRxiv 2020.08.13.249664; doi:https://doi.org/10.1101/2020.08.13.249664
[3]. Li H, Chang TW, et,al. Colcemid inhibits the rejoining of the nucleotide excision repair of UVC-induced DNA damages in Chinese hamster ovary cells. Mutat Res. 2005 Dec 30;588(2):118-28. doi: 10.1016/j.mrgentox.2005.09.005. Epub 2005 Nov 11. PMID: 16290038.
[4]. Tsutsui T, Maizumi H, et,al. Colcemid-induced neoplastic transformation and aneuploidy in Syrian hamster embryo cells. Carcinogenesis. 1984 Jan;5(1):89-93. doi: 10.1093/carcin/5.1.89. PMID: 6690091.
[5]. Bhuyan BK, Adams EG, et,al. Colcemid effects on B16 melanoma cell progression and aberrant mitotic division. J Cell Physiol. 1987 Aug;132(2):237-45. doi: 10.1002/jcp.1041320207. PMID: 3624316.
[6]. Nomura T. [In vivo cell cycle synchronization of the murine sarcoma 180 by continuous colcemid infusion (author's transl)]. Nihon Seikeigeka Gakkai Zasshi. 1980 Dec;54(12):1719-32. Japanese. PMID: 7288228.
[7]. Rieder CL, Palazzo RE. Colcemid and the mitotic cycle. J Cell Sci. 1992 Jul;102 ( Pt 3):387-92. doi: 10.1242/jcs.102.3.387. PMID: 1506421.
| Cas No. | 477-30-5 | SDF | |
| 别名 | 秋水仙碱,Demecolcine | ||
| 化学名 | (7S)-6,7-dihydro-1,2,3,10-tetramethoxy-7-(methylamino)-benzo[a]heptalen-9(5H)-one | ||
| Canonical SMILES | COC1=CC=C2C([C@@H](NC)CCC3=C2C(OC)=C(OC)C(OC)=C3)=CC1=O | ||
| 分子式 | C21H25NO5 | 分子量 | 371.4 |
| 溶解度 | 30mg/mL in ethanol, 25mg/mL in DMSO or DMF | 储存条件 | Store at -20°C |
| General tips | 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 储存时，请在 6 个月内使用，-20°C 储存时，请在 1 个月内使用。 为了提高溶解度，请将管子加热至37℃，然后在超声波浴中震荡一段时间。 |
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| Shipping Condition | 评估样品解决方案：配备蓝冰进行发货。所有其他可用尺寸：配备RT，或根据请求配备蓝冰。 | ||
| 制备储备液 | |||
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1 mg | 5 mg | 10 mg |
| 1 mM | 2.6925 mL | 13.4626 mL | 26.9251 mL |
| 5 mM | 0.5385 mL | 2.6925 mL | 5.385 mL |
| 10 mM | 0.2693 mL | 1.3463 mL | 2.6925 mL |
| 第一步：请输入基本实验信息（考虑到实验过程中的损耗，建议多配一只动物的药量） | ||||||||||
| 给药剂量 | mg/kg |  |
动物平均体重 | g | |
每只动物给药体积 | ul | |
动物数量 | 只 |
| 第二步：请输入动物体内配方组成（配方适用于不溶于水的药物；不同批次药物配方比例不同，请联系GLPBIO为您提供正确的澄清溶液配方） | ||||||||||
| % DMSO % % Tween 80 % saline | ||||||||||
| 计算重置 | ||||||||||
计算结果:
工作液浓度： mg/ml；
DMSO母液配制方法： mg 药物溶于 μL DMSO溶液（母液浓度 mg/mL，
体内配方配制方法：取 μL DMSO母液，加入 μL PEG300，混匀澄清后加入μL Tween 80，混匀澄清后加入 μL saline，混匀澄清。
1. 首先保证母液是澄清的；
2.
一定要按照顺序依次将溶剂加入，进行下一步操作之前必须保证上一步操作得到的是澄清的溶液，可采用涡旋、超声或水浴加热等物理方法助溶。
3. 以上所有助溶剂都可在 GlpBio 网站选购。
Effect of Colcemid on the centrosome and microtubules in dermal melanophores of Xenopus laevis larvae in vivo
Cell Mol Biol (Noisy-le-grand) 1999 Nov;45(7):1099-117.PMID:10644015doi
An electron microscopy study showed that in melanophores with dispersed and aggregated pigment the sensitivity of the centrosome and the stability of microtubules were different and depended on the Colcemid concentration. The structure of the centrosome didn't change upon exposure to Colcemid in dispersed melanophores. In aggregated melanophores, on exposure to 10(-6) M Colcemid, the centrosome retained its structure; Colcemid at 10(-5)-10(-3) M caused a dramatic collapse of the centrosome. Treatment of aggregated melanophores with Colcemid resulted in the complete disassembly of the microtubules; though microtubules in dispersed melanophores appear to be Colcemid resistant. Light microscopy studies indicated that in Xenopus melanophores with aggregated or dispersed pigment melanosomes didn't change their location after exposure to 10(-3)-10(-6) M Colcemid. Subsequent incubation in colcemid-free medium revealed that the cells retained their ability to translocate melanosomes in response to hormone stimulation. Electron microscopy data revealed the inactivation of the centrosome as MTOC (microtubule-organizing center) in dispersed melanophores with melatonin substituted for MSH in the presence of Colcemid. In contrast, with melanocyte-stimulating hormone (MSH) substituted for melatonin, we observed the activation of the centrosome in aggregated cells. We showed that in aggregated melanophores pigment movement proceeded in the complete absence of microtubules, suggesting the involvement of a microtubule-independent component in the hormone-induced melanosome dispersion. However, we observed abnormal aggregation along colcemid-resistent microtubules in dispersed melanophores, suggesting the involvement of not only stable but also labile microtubules in the centripetal movement of melanosomes. The results raise the intriguing questions about the mechanism of the hormone and Colcemid action on the centrosome structure and microtubule network in melanophores with dispersed and aggregated pigment.
Investigation of sphingosine kinase 1 inhibitory potential of cinchonine and Colcemid targeting anticancer therapy
J Biomol Struct Dyn 2022 Sep;40(14):6350-6362.PMID:33565370DOI:10.1080/07391102.2021.1882341.
Sphingosine kinase 1 (SphK1) and sphingosine-1-phosphate (S1P) signaling regulates numerous diseases such as cancer, diabetes, and inflammation-related ailments, rheumatoid arthritis, atherosclerosis, and multiple sclerosis. The importance of SphK1 in chemo-resistance has been extensively explored in breast, lung, colon, and hepatocellular carcinomas. SphK1 is considered an attractive drug target for the development of anticancer therapy. New drug molecules targeting the S1P signaling are required owing to its pleiotropic nature and association with multiple downstream targets. Here, we have investigated the binding affinity and SphK1 inhibitory potential of cinchonine and Colcemid using a combined molecular docking and simulation studies followed by experimental analysis. These compounds bind to SphK1 with a significantly high affinity and subsequently inhibit kinase activity (IC50 7-9 μM). Further, MD simulation studies revealed that both cinchonine and Colcemid bind to the residues at the active site pocket of SphK1 with several non-covalent interactions, which may be responsible for inhibiting its kinase activity. Besides, the binding of cinchonine and Colcemid causes substantial conformational changes in the structure of SphK1. Taken together, cinchonine and Colcemid may be implicated in designing potential drug molecules with improved affinity and specificity for SphK1 targeting anticancer therapy.Communicated by Ramaswamy H. Sarma.
Colcemid treatment during oocyte maturation improves preimplantation development of cloned pig embryos by influencing meiotic progression and cytoplasmic maturation
Mol Reprod Dev 2015 Jun;82(6):489-97.PMID:25982990DOI:10.1002/mrd.22498.
The objective of this study was to examine the effects of Colcemid treatment during oocyte in vitro maturation (IVM) and embryonic development after parthenogenetic activation (PA) and somatic-cell nucleus transfer (SCNT) in pigs. Immature oocytes were treated with Colcemid from 0 to 22, 38 to 42, or 0 to 22 hr followed by 38 to 42 hr during IVM (designated as COL0-22, COL38-42, and COL0-22/38-42, respectively). The proportion of oocytes reaching the germinal vesicle (GV)/GV breakdown (GVBD) stage after 22 hr of IVM was higher in COL0-22 (98.4%) than in controls not exposed to Colcemid (68.7%). The proportion of metaphase-II (MII) oocytes after 30 hr of IVM was higher in control (79.6%) than in COL0-22 oocytes (61.7%); overall nuclear progression to the MII stage was not influenced by Colcemid treatment by the end of the IVM period (93.8, 86.7, 86.8, and 84.8% for control, COL0-22, COL38-42, and COL0-22/38-42, respectively). COL0-22 oocytes showed higher intra-oocyte glutathione content (1.7 vs. 1.0-1.3 pixels/oocyte) and increased blastocyst formation after PA (68.7% vs. 42.5-52.2%) and SCNT (39.4% vs. 16.3-28.6%) than control, COL38-42, and COL0-22/38-42 oocytes. Colcemid treatment for 0-22 and 0-22/38-42 hr of IVM also stimulated the expression of cyclin-dependent kinase 1 (CDK1), proliferating cell nuclear antigen (PCNA), and extracellular signal-regulated kinase 2 (ERK2) mRNAs. Our results thus demonstrate that the presence of Colcemid during the early stage of IVM stimulates preimplantation development of PA and SCNT porcine embryos by improving the cytoplasmic microenvironment.
A fluorescent analog of Colcemid, N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)-colcemid, as a probe for the colcemid-binding sites of tubulin and microtubules
J Biol Chem 1987 May 5;262(13):6318-22.PMID:3571259doi
The synthesis and biological testing of the fluorescent analog of Colcemid, N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)-colcemid (NBD-colcemid), are here described. NBD-colcemid exhibited a visible absorption maximum at 465 nm and fluoresced in the range of 520-540 nm, highly in environments of low polarity, whereas only slightly in aqueous solution. The addition of NBD-colcemid to bovine brain tubulin was accompanied by a striking enhancement of fluorescence. The fluorescent titration study suggested a stoichiometric binding of NBD-colcemid to tubulin. Assembled microtubules were directly visualized after mixing with NBD-colcemid using a fluorescence microscope. NBD-colcemid reversibly disrupted the metaphase spindles of sea urchin eggs as well as unlabeled Colcemid. However, even when the birefringence of spindles was mostly lost, self-quenching properties of the NBD fluorescence allowed tubulin and its oligomers aggregated in higher concentrations in eggs to be visualized under a fluorescence microscope. The results suggest a wide applicability of NBD-colcemid as a fluorescent probe for studying the interactions of Colcemid with tubulin and microtubules, as well as for localizing other colcemid-binding structures within cells.
Colcemid inhibits the rejoining of the nucleotide excision repair of UVC-induced DNA damages in Chinese hamster ovary cells
Mutat Res 2005 Dec 30;588(2):118-28.PMID:16290038DOI:10.1016/j.mrgentox.2005.09.005.
In our previous study, we found that Colcemid, an inhibitor of mitotic spindle, promotes UVC-induced apoptosis in Chinese hamster ovary cells (CHO.K1). In this study, a brief treatment of Colcemid on cells after but not before UV irradiation could synergistically reduce the cell viability. Although Colcemid did not affect the excision of UV-induced DNA damages such as [6-4] photoproducts or cyclobutane pyrimidine dimers, Colcemid accumulated the DNA breaks when it was added to cells following UV-irradiation. This Colcemid effect required nucleotide excision repair (NER) since the same accumulation of DNA breaks was barely or not detected in two NER defective strains of CHO cells, UV5 or UV24. Furthermore, the Colcemid effect was not due to semi-conservative DNA replication or mitosis since the colcemid-caused accumulation of DNA breaks was also seen in non-replicating cells. Moreover, Colcemid inhibited rejoining of DNA breaks accumulated by hydroxyurea/cytosine arabinoside following UV irradiation. Nevertheless, Colcemid did not affect the unscheduled DNA synthesis as assayed by the incorporation of bromodeoxyuridine. Taken together, our results suggest that Colcemid might inhibit the step of ligation of NER pathways.