Yonsei Med J.  2017 Mar;58(2):272-281. 10.3349/ymj.2017.58.2.272.

Comparative Effects of Non-Thermal Atmospheric Pressure Plasma on Migration and Invasion in Oral Squamous Cell Cancer, by Gas Type

• Kang SU
• Seo SJ
• Kim YS
• Shin YS
• Koh YW
• Lee CM
• Yang SS
• Lee JS
• Moon E
• Kang H
• Ryeo JB
• Lee Y
• Kim CH

• KMID: 2427114
• DOI: http://doi.org/10.3349/ymj.2017.58.2.272

Abstract

PURPOSE
The fourth state of matter, plasma is known as an ionized gas with electrons, radicals and ions. The use of non-thermal plasma (NTP) in cancer research became possible because of the progresses in plasma medicine. Previous studies on the potential NTP-mediated cancer therapy have mainly concentrated on cancer cell apoptosis. In the present study, we compared the inhibitory effect of NTP on cell migration and invasion in the oral squamous cancer cell lines.
MATERIALS AND METHODS
We used oral squamous cancer cell lines (SCC1483, MSKQLL1) and different gases (Nâ‚‚, He, and Ar). To investigate the mechanism of plasma treatment, using different gases (Nâ‚‚, He, and Ar) which induces anti-migration and anti-invasion properties, we performed wound healing assay, invasion assay and gelatin zymography.
RESULTS
The results showed that NTP inhibits cancer cell migration and invasion of oral squamous cancer cell. In addition, focal adhesion kinase expression and matrix metalloproteinase-2/9 activity were also inhibited.
CONCLUSION
The suppression of cancer cell invasion by NTP varied depending on the type of gas. Comparison of the three gases revealed that Nâ‚‚ NTP inhibited cell migration and invasion most potently via decreased expression of focal adhesion kinase and matrix metalloproteinase activity.

Keyword

Non thermal atmospheric pressure plasma; oral squamous cancer cell; focal adhesion kinase; matrix metallopeptidase-2/9; paxillin

MeSH Terms

Apoptosis/drug effects
Carcinoma, Squamous Cell/*metabolism
Cell Line, Tumor
Cell Movement/*drug effects
Focal Adhesion Protein-Tyrosine Kinases
Humans
Matrix Metalloproteinase 2/genetics/*metabolism
Matrix Metalloproteinase 9/genetics/*metabolism
Mouth Neoplasms/metabolism/*therapy
Neoplasm Invasiveness
Plasma Gases/*pharmacology
Wound Healing
Plasma Gases
Focal Adhesion Protein-Tyrosine Kinases
Matrix Metalloproteinase 2
Matrix Metalloproteinase 9

Figure

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  Fig. 1 Optical emission spectrum (OES) of plasma jet according to gases (A) OES of NTP induced by N2 gas, (B) He gas, and (C) Ar gas. NTP, non-thermal plasma.

• 

  Fig. 2 Cell death after 1, 3, and 5 min NTP treatment according to gases (N2, He, or Ar). (A) Annexin V-PI assay and quantification of SCC1483. Bar graph: mean±SD of 3 independent experiments. *p<0.05, ***p<0.001. NTP, non-thermal plasma; PI, propidium iodide. *p<0.05. NTP, non-thermal plasma; PI, propidium iodide.

• 

  Fig. 3 Wound healing assay of 3 min NTP treatment according to gases (N2, He, or Ar) on (A) SCC1483 and (B) MSKQLL1. Data represents mean±SD of three independent experiments. *p<0.05, ***p<0.001. NTP, non-thermal plasma.

• 

  Fig. 4 Invasion assay of 3 min NTP treatment according to gases (N2, He, or Ar) on SCC1483 (A) and MSKQLL1 (B). Data represent mean±SD of three independent experiments. **p<0.01, ***p<0.001. NTP, non-thermal plasma. (C) Gelatin zymography of 3 min NTP treatment according to gases (N2, He, or Ar) for MMP-2/9 on SCC1483 and MSKQLL1. *p<0.05, ***p<0.001. NTP, non-thermal plasma; MMPs, matrix metalloproteinases.

• 

  Fig. 5 (A) Western blots of p-focal adhesion kinase (FAK), FAK, Integrin (b3), p-paxillin, paxillin, and uncleaved caspase-3 after 3 min NTP treatment according to gases (N2, He, or Ar). (B) Immunocytochemical assay for p-FAK after 3 min NTP treatment according to gases (N2, He, or Ar) in SCC1483 and (C) MSKQLL1. Scale bar=10 µm. Each figure is representative of three experiments with triplicates. NTP, non-thermal plasma.

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