Microtubules, part of the cytoskeleton that provides structure and shape to the cytoplasm of cells, are tubular polymers of tubulin formed by polymerization. Nocodazole is a standard anti-microtubule agent that interferes with the polymerization and now be mostly used as depolymerizing drug in basic cell biology research[8]. Nocodazole can be used as a high-affinity ligand for the cancer-related kinases including Abl phosphorylated, c-Kit, BRAF, and MEK. Furthermore, Nocodazole induces apoptosis in chronic lymphocytic leukemia cells, inhibits insulin-stimulated glucose transport and decreases apoptosis in some human colon carcinoma cells[9]. In vitro Tubulin Polymerization Assays, Nocodazole inhibited tumor cell lines including NCIH460, SKOV3, BT549,451LU, COLO-205 at 48h with IC50 values 68nM, 80nM, 69nM, 72nM, 88nM respectively, which was reflected by a decreased level of steady state turbidity in a dose-dependent manner[5]. To learn how the spindle assembly checkpoint (SAC) inhibitory is extinguished, 9A-Hec1-expressing cells exhibited a robust mitotic arrest after incubation in 5 mM nocodazole for 5 h. As comparison, WT-Hec1 was treated with a moderately low dose (300 nM) of nocodazole, which resulted in the loss of all non-kinetochore spindle microtubules but retention of kinetochore–microtubule[6]. In addition, nocodazole can also improve the CRISPR-mediated HDR efficiency, a 600-bp homology in both arms leads to high-level genome knock in, with 97–100% of the donor insertion events being mediated by HDR. The combined use of CCND1, a cyclin that functions in G1/S transition, and nocodazole, a G2/M phase synchronizer, doubles HDR efficiency to up to 30% in iPSCs, the results showed an additive effect on enhancing precise genome editing[7].
作用机制
Nocodazole will bind to β-Tb and block polymerization, arrest mammalian cells in mitosis and release them into cytokinesis and G1 as well as mis-regulate mitotic kinases[8].
Nocodazole/诺考达唑 细胞实验
Cell Line
Concentration
Treated Time
Description
References
TLX1-T-ALL lymphoblasts
100 ng/ml
24 h
To test the function of the mitotic checkpoint in TLX1-induced leukemias, the results showed that TLX1 mouse T-ALL lymphoblasts fail to arrest in mitosis following disruption of the mitotic spindle
Nat Med. 2010 Nov;16(11):1321-7.
Mouse embryonic stem cells
50 ng/ml
6 h
Used for synchronizing mouse embryonic stem cells, mitotic cells were collected via shake-off.
Nucleic Acids Res. 2023 Jun 9;51(10):5040-5055.
mouse cortical neurons
6 μM
30 min
To test the effect of Cep120 overexpression on microtubule stability, it was found that Cep120 overexpression protected MTs against the depolymerization effect of nocodazole.
Cell Rep. 2022 Apr 19;39(3):110686.
SN neurons
45 nM
12 h
To simulate ICH and microtubule depolymerization, and observe its effects on neurons.
J Am Heart Assoc. 2018 Jan 18;7(2):e007626.
MCF7 cells
10 µM
4 h
To study the effect of nocodazole on focal adhesion (FA) formation and disassembly in MCF7 cells. Results showed that nocodazole treatment increased FA numbers, followed by rapid disassembly and reassembly after nocodazole washout.
Br J Cancer. 2009 Feb 24;100(4):633-43.
MDA231-ErbB2 cells
10 µM
4 h
To study the effect of nocodazole on focal adhesion (FA) formation and disassembly in MDA231-ErbB2 cells. Results showed that nocodazole treatment increased FA numbers, followed by rapid disassembly and reassembly after nocodazole washout.
Br J Cancer. 2009 Feb 24;100(4):633-43.
HSCs
10 nM
24 h
To determine the type of HSC division, results showed that G5/G6 HSCs had a lower frequency of symmetric self-renewal division and a higher frequency of symmetric commitment division.
Nat Commun. 2021 Nov 25;12(1):6850.
Nocodazole/诺考达唑 动物实验
Species
Animal Model
Administration
Dosage
Frequency
Description
References
C57/BL6 mice
ICH model
Intrastriatal injection
100 μM(10 μL)
Single injection, lasting 7 days
To simulate the pathological characteristics of ICH and observe the effects of nocodazole on motor function and dopamine neurons.
J Am Heart Assoc. 2018 Jan 18;7(2):e007626.
Nude mice
HCT116 cell xenograft model
10 mg/kg
Every 2 days for 10 days
Nocodazole inhibited tumor growth by inducing LATS2 to suppress β-catenin/BCL9-mediated transcription.
Cell Rep. 2013 Dec 26;5(6):1650-63
Mice
Newborn mice
Subcutaneous injection
0.4 mM
Single injection
To study the effect of cortical microtubules on tight junction function in vivo. The results showed that disruption of microtubules led to loss of tight junction function, increasing the diffusion of biotin.
Antiproliferative activity against human 451LU cells after 48 hrs by alamar blue assay, IC50=72 nM
21563750
human A549 cells
Proliferation assay
48 h
Antiproliferative activity against human A549 cells after 48 hrs by SRB assay, GI50=20 nM
25827522
human A549 cells
Cytotoxic assay
24 h
Cytotoxicity against human A549 cells after 24 hrs by MTT assay, IC50=36.6 nM
20110137
human A549 cells
Growth inhibition assay
Growth inhibition of human A549 cells by Sulforhodamine B assay, GI50=0.1 μM
21106458
human BT549 cells
Proliferation assay
48 h
Antiproliferative activity against human BT549 cells after 48 hrs by alamar blue assay, IC50=69 nM
21563750
human COLO205 cells
Proliferation assay
48 h
Antiproliferative activity against human COLO205 cells after 48 hrs by alamar blue assay , IC50=88 nM
21563750
human HaCaT cells
Cytotoxic assay
48 h
Cytotoxic activity against human HaCaT cells assessed as reduction in cell growth incubated for 48 hrs by MTT assay, IC50=8.9 μM
26231080
human HeLa cells
Proliferation assay
48 h
Antiproliferative activity against human HeLa cells after 48 hrs by SRB assay, GI50=15 nM
25827522
human Jurkat T cells
Function assay
36 h
Induction of microtubule damage in human Jurkat T cells overexpressing Bcl2 after 36 hrs by immunostaining method
22197393
human KB/HeLa cells
Function assay
24 h
Cell cycle arrest in human KB/HeLa cells assessed as G2/M phase accumulation after 24 hrs by FACS analysis, EC50=91 nM
19220018
human MCF7 cells
Growth inhibition assay
Growth inhibition of human MCF7 cells by Sulforhodamine B assay, GI50=0.1 μM
21106458
human MDA-MB-231 cells
Proliferation assay
48 h
Antiproliferative activity against human MDA-MB-231 cells after 48 hrs by SRB assay, GI50=17 nM
25827522
human MIAPaCa2 cells
Proliferation assay
48 h
Antiproliferative activity against human MIAPaCa2 cells assessed as growth inhibition after 48 hrs by SRB assay, GI50=0.95 μM
25131956
human NCI-H460 cells
Proliferation assay
48 h
Antiproliferative activity against human NCI-H460 cells after 48 hrs by alamar blue assay, IC50=68 nM
21563750
human NCI-H460 cells
Proliferation assay
48 h
Antiproliferative activity against human NCI-H460 cells after 48 hrs by XTT assay, IC50=0.15 μM
20537765
human PANC1 cells
Proliferation assay
48 h
Antiproliferative activity against human PANC1 cells after 48 hrs by SRB assay, GI50=10 nM
25827522
human PC3M cells
0.5 ug/ml
Function assay
24 h
Induction of apoptosis in human PC3M cells assessed as accumulation at subG1 phase at 0.5 ug/ml after 24 hrs by flow cytometry
18247573
human RKO cells
Proliferation assay
Antiproliferative activity against human RKO cells by XTT assay, IC50=0.11 μM
19220018
human SF268 cells
Cytotoxic assay
48 h
Cytotoxicity against human SF268 cells after 48 hrs by XTT assay, IC50=0.3 μM
21705223
human SGC7901 cells
Function assay
48 h
Induction of distortions in tubulin assembly in human SGC7901 cells assessed as abnormal mitotic spindle formation at 2 fold IC50 level incubated for 48 hrs by immunofluorescence staining based fluorescence microscopy
25262051
human SKBR3 cells
Growth inhibition assay
48 h
Growth inhibition of human SKBR3 cells after 48 hrs by MTT assay
22850214
human SK-N-SH cells
Growth inhibition assay
Growth inhibition of human SK-N-SH cells by Sulforhodamine B assay, GI50=0.66 μM
21106458
human SKOV3 cells
Proliferation assay
48 h
Antiproliferative activity against human SKOV3 cells after 48 hrs by alamar blue assay, IC50=80 nM
21563750
human SW480 cells
Proliferation assay
48 h
Antiproliferative activity against human SW480 cells after 48 hrs by alamar blue assay, IC50=0.12 μM
21563750
human U2OS cells
Function assay
72 h
Induction of mitotic arrest in human U2OS cells assessed as mitotic index after 72 hrs by microscopic analysis, EC50=50 nM
24900887
K562 cells
Function assay
Effective concentration required to arrest propidium iodide stained K562 cells in G2/M phase, EC50=82 nM
12852768
L1210 murine leukemia cell
Growth inhibition assay
In vitro for its inhibitory activity against L1210 murine leukemia cell growth, IC50=38 nM
1738146
LT12 cells
Proliferation assay
48 h
Antiproliferative activity against LT12 cells by XTT assay after 48 hrs, IC50=40 nM
17181164
lymphoid leukemia L1210 cells
Proliferation assay
48 h
Inhibitory concentration against proliferation of cultured lymphoid leukemia L1210 cells during 48 hrs, ID50=27 nM
7131483
mouse NIH3T3 cells
0.5 ug/ml
Function assay
24 h
Induction of apoptosis in mouse NIH3T3 cells assessed as accumulation at subG1 phase at 0.5 ug/ml after 24 hrs by flow cytometry
18247573
mouse P388 cells
Proliferation assay
48 h
Antiproliferative activity against mouse P388 cells after 48 hrs by XTT assay, IC50=40 nM
19220018
rat LT12 cells
Proliferation assay
48 h
Antiproliferative activity against rat LT12 cells after 48 hrs by XTT assay, IC50=6 nM
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