Research ArticleBIOPHYSICS

Magnetism and photo dual-controlled supramolecular assembly for suppression of tumor invasion and metastasis

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Science Advances  19 Sep 2018:
Vol. 4, no. 9, eaat2297
DOI: 10.1126/sciadv.aat2297
  • Fig. 1 Schematic illustration of the formation of MitP-MNP⊂HACD nanofibers.
  • Fig. 2 Formation and characterization of MitP-MNP and MitP-MNP⊂HACD nanofibers.

    (A) Schematic illustration of formation of MitP-MNP⊂HACD nanofibers by cross-linking HACD with MitP-MNP. (B and C) TEM images and (D) dynamic light scattering (DLS) data for MitP-MNP and MitP-MNP⊂HACD nanofibers ([MitP-MNP] = 0.2 mg/ml and [HACD] = 0.2 mg/ml). (E) 1H nuclear magnetic resonance (NMR) titration of MitP with native β-CD, revealing that the cyclohexyl groups of MitP were included in the β-CD cavity: MitP alone (i) and [MitP]:[β-CD] [1:1, 1:2, 1:3, and 1:4 (ii to v), respectively]. ppm, parts per million.

  • Fig. 3 Magnetism- and photo-controlled assembly of MitP-MNP⊂HACD nanofibers.

    (A and B) Confocal microscopy images of the growth of MitP-MNP⊂HACD nanofibers along the direction of the geomagnetic field (0.050 mT). (C and D) Light microscopy images of the growth of MitP-MNP⊂HACD nanofibers in a pure geomagnetic field (0.048 mT; C) and in a metal-caged room with a decreased geomagnetic field (0.015 mT; D). (E) Confocal microscopy images of the growth of MitP-MNP⊂HACD nanofibers along the direction of an artificial magnetic field (0.308 mT). (F) Schematic illustration of photoresponsive assembly and disassembly of MitP-MNP⊂HACD nanofibers in the presence of an arylazopyrazole (AAP) carboxylate ([MitP-MNP] = 0.2 mg/ml, [HACD] = 0.2 mg/ml, and [AAP] = 0.2 mg/ml). Vis, visible. (G) Confocal microscopy images showing the temporal dependence of nanofiber growth in the presence of trans-AAP and cis-AAP carboxylates [generated by ultaviolet (UV) irradiation at 365 nm].

  • Fig. 4 Interaction of MitP-MNP⊂HACD nanofibers with mitochondria and disruption of mitochondrial function in A549 tumor cells by the nanofibers.

    (A) Schematic illustration of aggregation of mitochondria around MitP-MNP⊂HACD nanofibers along the direction of the geomagnetic field (0.050 mT). (B) Confocal microscopy images of DAPI-labeled mitochondria incubated with MitP-MNP and HACD for 2, 5, and 10 min ([MitP-MNP] = 0.2 mg/ml and [HACD] = 0.2 mg/ml). (C) Confocal microscopy images of cells treated with MitP-MNP alone or with MitP-MNP⊂HACD nanofibers (white arrows indicate intracellular nanofibers). (D) Images of Western blots of cytoplasmic and mitochondrial cytochrome C [Cyt C(cyto) and Cyt C(mit)], cleaved caspase-3 (an apoptosis-inducing factor), and tubulin. (E) Ratio of Cyt C(cyto) to Cyt C(mit). (F) Ratio of cleaved caspase-3 to tubulin. (G) Decrease of intracellular adenosine 5′-triphosphate (ATP) production caused by MitP-MNP⊂HACD nanofibers. (H) Decrease in cell viability caused by MitP-MNP⊂HACD nanofibers. (I) Cell cycle arrest and apoptosis caused by MitP-MNP⊂HACD nanofibers. Asterisks in E to H indicate statistically significant differences between groups (P < 0.05).

  • Fig. 5 Suppression of tumor cell invasion and metastasis by MitP-MNP⊂HACD nanofibers both in vitro and in vivo.

    (A) Schematic illustration of inhibition of tumor cell invasion into Matrigel by MitP-MNP⊂HACD nanofibers. (B) Light microscopy images of the interaction between RFP-tagged A549 lung tumor cells and MitP-MNP⊂HACD nanofibers during invasion into the Matrigel. (C) Confocal microscopy images of the invading tumor cells at various heights in the Matrigel. (D) Quantification of the fluorescence intensity of the tumor cells at various heights in the Matrigel. (E) Images of migration of the A549 cells to the scraped gaps. (F) In vivo inhibition of A549 cell metastasis by MitP-MNP⊂HACD nanofibers (dotted blue circles indicate tumor cell injection sites). (G) Survival curve for mice with A549 metastases; the black arrow indicates the time at which MitP-MNP or MitP-MNP⊂HACD nanofibers were injected into the mice.

Supplementary Materials

  • Supplementary material for this article is available at http://advances.sciencemag.org/cgi/content/full/4/9/eaat2297/DC1

    Table S1. Angles between the magnetic field line and the growth direction of supramolecular nanofibers and the growth rate of the MitP-MNP⊂HACD assembly under different magnetic fields.

    Fig. S1. Synthetic route of MitP-MNP.

    Fig. S2. Characterization of the synthesized MNP, MitP, and MitP-MNP.

    Fig. S3. Effect of HA, 1-adamantanecarboxylic acid, and arylazopyrazole on assembly of the nanofibers.

    Fig. S4. Effect of ADA and artificial magnetic field on aggregation of mitochondria along with the direction of external magnetic field.

    Fig. S5. Amplified confocal microscopy images of the A549 cells treated by MitP-MNP or MitP-MNP⊂HACD.

    Fig. S6. Localization of ABP-MNP as negative control.

    Fig. S7. Effect of MitP-MNP, MitP-MNP⊂HACD, and HACD on cell viability and cell death.

    Fig. S8. Confocal microscopy images of MitP-MNP⊂HACD in the Matrigel invasion model.

    Fig. S9. Binding activity of MitP-MNP⊂HACD with the tumor cells (A549-RFP) and the normal cells (293T) and the impact of the nanofibers on the RES organs.

    Movie S1. Confocal microscopic observation of MitP-MNP and HACD aggregate in the geomagnetism field.

    Movie S2. Confocal microscopic observation of MitP-MNP and HACD aggregate in the geomagnetism field with different sample locations.

    Movie S3. Microscopic observation of MitP-MNP and HACD aggregate upon deviation from Earth’s magnetic field line.

    Movie S4. Microscopic observation of MitP-MNP and HACD aggregate with a canceling system for artificial magnetism fields.

    Movie S5. Microscopic observation of MitP-MNP and HACD aggregate in a metal-caged room.

    Movie S6. Microscopic observation of MitP-MNP and HACD aggregate in an artificial magnetic field (0.308 mT).

    Movie S7. Microscopic observation of MitP-MNP and HACD aggregate in an artificial magnetic field (2.968 mT).

  • Supplementary Materials

    The PDF file includes:

    • Table S1. Angles between the magnetic field line and the growth direction of supramolecular nanofibers and the growth rate of the MitP-MNP⊂HACD assembly under different magnetic fields.
    • Fig. S1. Synthetic route of MitP-MNP.
    • Fig. S2. Characterization of the synthesized MNP, MitP, and MitP-MNP.
    • Fig. S3. Effect of HA, 1-adamantanecarboxylic acid, and arylazopyrazole on assembly of the nanofibers.
    • Fig. S4. Effect of ADA and artificial magnetic field on aggregation of mitochondria along with the direction of external magnetic field.
    • Fig. S5. Amplified confocal microscopy images of the A549 cells treated by MitP-MNP or MitP-MNP⊂HACD.
    • Fig. S6. Localization of ABP-MNP as negative control.
    • Fig. S7. Effect of MitP-MNP, MitP-MNP⊂HACD, and HACD on cell viability and cell death.
    • Fig. S8. Confocal microscopy images of MitP-MNP⊂HACD in the Matrigel invasion model.
    • Fig. S9. Binding activity of MitP-MNP⊂HACD with the tumor cells (A549-RFP) and the normal cells (293T) and the impact of the nanofibers on the RES organs.

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    Other Supplementary Material for this manuscript includes the following:

    • Movie S1 (.mp4 format). Confocal microscopic observation of MitP-MNP and HACD aggregate in the geomagnetism field.
    • Movie S2 (.mp4 format). Confocal microscopic observation of MitP-MNP and HACD aggregate in the geomagnetism field with different sample locations.
    • Movie S3 (.mp4 format). Microscopic observation of MitP-MNP and HACD aggregate upon deviation from Earth’s magnetic field line.
    • Movie S4 (.mp4 format). Microscopic observation of MitP-MNP and HACD aggregate with a canceling system for artificial magnetism fields.
    • Movie S5 (.mp4 format). Microscopic observation of MitP-MNP and HACD aggregate in a metal-caged room.
    • Movie S6 (.mp4 format). Microscopic observation of MitP-MNP and HACD aggregate in an artificial magnetic field (0.308 mT).
    • Movie S7 (.mp4 format). Microscopic observation of MitP-MNP and HACD aggregate in an artificial magnetic field (2.968 mT).

    Files in this Data Supplement:

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