Cell migration inhibition activity of a non-RGD disintegrin from Crotalus durissus collilineatus venom View Full Text


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Article Info

DATE

2018-10-20

AUTHORS

Isadora Sousa de Oliveira, Rafaella Varzoni Manzini, Isabela Gobbo Ferreira, Iara Aimê Cardoso, Karla de Castro Figueiredo Bordon, Ana Rita Thomazela Machado, Lusânia Maria Greggi Antunes, José Cesar Rosa, Eliane Candiani Arantes

ABSTRACT

BackgroundIn recent decades, snake venom disintegrins have received special attention due to their potential use in anticancer therapy. Disintegrins are small and cysteine-rich proteins present in snake venoms and can interact with specific integrins to inhibit their activities in cell-cell and cell-ECM interactions. These molecules, known to inhibit platelet aggregation, are also capable of interacting with certain cancer-related integrins, and may interfere in important processes involved in carcinogenesis. Therefore, disintegrin from Crotalus durissus collilineatus venom was isolated, structurally characterized and evaluated for its toxicity and ability to interfere with cell proliferation and migration in MDA-MB-231, a human breast cancer cell line.MethodsBased on previous studies, disintegrin was isolated by FPLC, through two chromatographic steps, both on reversed phase C-18 columns. The isolated disintegrin was structurally characterized by Tris-Tricine-SDS-PAGE, mass spectrometry and N-terminal sequencing. For the functional assays, MTT and wound-healing assays were performed in order to investigate cytotoxicity and effect on cell migration in vitro, respectively.ResultsDisintegrin presented a molecular mass of 7287.4 Da and its amino acid sequence shared similarity with the disintegrin domain of P-II metalloproteases. Using functional assays, the disintegrin showed low cytotoxicity (15% and 17%, at 3 and 6 μg/mL, respectively) after 24 h of incubation and in the wound-healing assay, the disintegrin (3 μg/mL) was able to significantly inhibit cell migration (24%, p < 0.05), compared to negative control.ConclusionThus, our results demonstrate that non-RGD disintegrin from C. d. collilineatus induces low cytotoxicity and inhibits migration of human breast cancer cells. Therefore, it may be a very useful molecular tool for understanding ECM-cell interaction cancer-related mechanisms involved in an important integrin family that highlights molecular aspects of tumorigenesis. Also, non-RGD disintegrin has potential to serve as an agent in anticancer therapy or adjuvant component combined with other anticancer drugs. More... »

PAGES

28

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URI

http://scigraph.springernature.com/pub.10.1186/s40409-018-0167-6

DOI

http://dx.doi.org/10.1186/s40409-018-0167-6

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https://app.dimensions.ai/details/publication/pub.1107740923

PUBMED

https://www.ncbi.nlm.nih.gov/pubmed/30377432


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22 schema:keywords BackgroundIn recent decades
23 ConclusionThus
24 Crotalus durissus collilineatus venom
25 DA
26 FPLC
27 MDA-MB-231
28 MTT
29 MethodsBased
30 N-terminal sequencing
31 RGD-disintegrins
32 SDS-PAGE
33 Tris-Tricine
34 ability
35 acid sequence
36 activity
37 adjuvant components
38 agents
39 aggregation
40 amino acid sequence
41 anticancer drugs
42 anticancer therapy
43 aspects
44 assays
45 attention
46 breast cancer cell lines
47 breast cancer cells
48 cancer cell lines
49 cancer cells
50 cancer-related mechanisms
51 carcinogenesis
52 cell lines
53 cell migration
54 cell proliferation
55 cell-ECM interactions
56 cells
57 chromatographic steps
58 collilineatus
59 collilineatus venom
60 column
61 components
62 control
63 cysteine-rich protein
64 cytotoxicity
65 decades
66 disintegrin
67 disintegrin domain
68 domain
69 drugs
70 effect
71 family
72 functional assays
73 human breast cancer cell lines
74 human breast cancer cells
75 important process
76 incubation
77 inhibition activity
78 integrin family
79 integrins
80 interaction
81 lines
82 low cytotoxicity
83 mass
84 mass spectrometry
85 mechanism
86 metalloproteases
87 migration
88 migration inhibition activity
89 molecular aspects
90 molecular mass
91 molecular tools
92 molecules
93 negative control
94 order
95 phase C
96 platelet aggregation
97 potential use
98 previous studies
99 process
100 proliferation
101 protein
102 recent decades
103 results
104 reversed phase C
105 sequence
106 sequencing
107 similarity
108 snake venom
109 snake venom disintegrins
110 special attention
111 specific integrins
112 spectrometry
113 step
114 study
115 therapy
116 tool
117 toxicity
118 tumorigenesis
119 use
120 useful molecular tool
121 venom
122 vitro
123 wound-healing assays
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