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SCIENCE CHINA Materials, Volume 60, Issue 3: 269-278(2017) https://doi.org/10.1007/s40843-016-5152-2

The dynamic interactions between chemotherapy drugs and plasmid DNA investigated by atomic force microscopy

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  • ReceivedOct 31, 2016
  • AcceptedDec 19, 2016
  • PublishedJan 13, 2017

Abstract

The advent of atomic force microscopy (AFM) provides a powerful tool for imaging individual DNA molecules. Chemotherapy drugs are often related to DNAs. Though many specific drug-DNA interactions have been observed by AFM, knowledge about the dynamic interactions between chemotherapy drugs and plasmid DNAs is still scarce. In this work, AFM was applied to investigate the nanoscale interactions between plasmid DNAs and two commercial chemotherapy drugs (methotrexate and cisplatin). Plasmid DNAs were immobilized on mica which was coated by silanes in advance. AFM imaging distinctly revealed the dynamic changes of single plasmid DNAs after the stimulation of methotrexate and cisplatin. Geometric features of plasmid DNAs were extracted from AFM images and the statistical results showed that the geometric features of plasmid DNAs changed significantly after the stimulation of drugs. This research provides a novel idea to study the actions of chemotherapy drugs against plasmid DNAs at the single-molecule level.


Funded by

National Natural Science Foundation of China(61503372,61522312,61327014,61433017)

the Youth Innovation Promotion Association CAS

and the CAS FEA International Partnership Program for Creative Research Teams.


Acknowledgment

This work was supported by the National Natural Science Foundation of China (61503372, 61522312, 61327014 and 61433017), the Youth Innovation Promotion Association CAS, and the CAS FEA International Partnership Program for Creative Research Teams.


Interest statement

The authors declare that there is no conflict of interests.


Contributions statement

Li M and Liu L designed the project. Li M performed the experiments and wrote the paper with support from Xi N and Wang Y. Xiao X was involved in data analysis. All authors contributed to the general discussion.


Author information

Mi Li is currently an associate professor at the State Key Laboratory of Robotics, Shenyang Institute of Automation, Chinese Academy of Sciences, Shenyang, China. His research interests include AFM-based imaging and mechanical analysis of biological systems.


Lianqing Liu is currently a professor at the State Key Laboratory of Robotics, Shenyang Institute of Automation, Chinese Academy of Sciences, Shenyang, China. His research interests include micro/nano system, nanodevice fabrication, nanobiotechnology and biosensors.


Ning Xi is currently a John D. Ryder Professor of Electrical and Computer Engineering at Michigan State University, East Lansing and a professor of Shenyang Institute of Automation, Chinese Academy of Sciences, Shenyang, China. His research interests include robotics, manufacturing automation, nanosensors, micro/nano manufacturing, and intelligent control and systems.


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  • Figure 1

    The experimental platform. (a) An actual photograph of AFM. (b) Optical image of the AFM cantilever. (c) Tuning the driving frequency of AFM cantilever.

  • Figure 2

    AFM imaging of plasmid DNA on mica. (a, b) AFM height images of bare mica (a) and plasmid DNAs on bare mica (b). (c) AFM height image of APTES-coated mica without DNA. (d, e) AFM height images of plasmid DNAs on APTES-coated mica (d, large size scan; e, small size scan of a single plasmid DNA). (f) Section curves taken along the red line in (b) and blue line in (e), respectively.

  • Figure 3

    AFM height images of single plasmid DNAs obtained at different scan forces: (a) 200 pN, (b) 350 pN, (c) 500 pN and their corresponding section curves taken along the red line in each image. (d, e) AFM height images under scan force of 100 pN. (f) Section curves taken along the red line in (d) and blue line in (e) respectively.

  • Figure 4

    AFM height images of single plasmid DNAs from the control group (without drugs) and extraction of geometric information. The red arrows denote the cross points.

  • Figure 5

    AFM height images of plasmid DNAs after the stimulation of methotrexate: (a, b) 1 ng μL−1, 30 min; (c–e) 5 ng μL−1, 30 min.

  • Figure 6

    Dynamic changes of supercoil plasmid DNAs after the stimulation of methotrexate. (a, b) Control group without methotrexate. The arrows in (a, b) indicate the supercoil structures, and the insets in are the section curves taken along the red lines. (c, d) 5 ng μL−1 methotrexate, 30 min. (e, f) 5 ng μL−1 methotrexate, 60 min. The arrows in (e, f) indicate the linear DNAs.

  • Figure 7

    AFM height images of plasmid DNAs after the stimulation of cisplatin. (a–e) 5 ng μL−1, 30 min. (f–h) 5 ng μL−1, 60 min. (i–k) Section curves taken along the red lines in (f–h), respectively.

  • Figure 8

    Changes of geometric features of plasmid DNAs after the stimulation of chemotherapy drugs (a, methotrexate; b, cisplatin) for different drug concentrations and incubation times. The values of length and width are shown as mean±SEM.

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