Tissue Tolerable Plasma (TTP) induces apoptosis in pancreatic cancer cells in vitro and in vivo

BMC Cancer

Lars Ivo Partecke¹  Katja Evert⁴  Jan Haugk¹  Friderike Doering¹  Lars Normann¹  Stephan Diedrich¹  Frank-Ulrich Weiss³  Matthias Evert⁴  Nils Olaf Huebner⁵  Cristin Guenther¹  Claus Dieter Heidecke¹  Axel Kramer⁵  René Bussiahn²  Klaus-Dieter Weltmann²  Onur Pati¹  Claudia Bender⁵  Wolfram von Bernstorff¹ 

[1] Department of General, Visceral, Thoracic and Vascular Surgery, University Medicine Greifswald, Ernst-Moritz-Arndt University, Greifswald, Germany

[2] Leibniz Institute of Plasma Science and Technology (INP), Felix Hausdorff-Str. 2, Greifswald, Germany

[3] Department of Gastroenterology and Nephrology, University Medicine Greifswald, Ernst-Moritz-Arndt University, Greifswald, Germany

[4] Institute of Pathology, University Medicine Greifswald, Ernst-Moritz-Arndt University, Greifswald, Germany

[5] Institute of Hygiene and Environmental Medicine, University Medicine Greifswald, Ernst-Moritz-Arndt University, Greifswald, Germany

关键词: Pancreatic cancer;    TUM-CAM Assay;    Apoptosis;    Plasma medicine;    Tissue tolerable plasma (TTP);   

Others  :  1080137
DOI  :  10.1186/1471-2407-12-473

 received in 2012-04-01, accepted in 2012-10-10,  发布年份 2012

【 摘 要 】

Background

The rate of microscopic incomplete resections of gastrointestinal cancers including pancreatic cancer has not changed considerably over the past years. Future intra-operative applications of tissue tolerable plasmas (TTP) could help to address this problem. Plasma is generated by feeding energy, like electrical discharges, to gases. The development of non-thermal atmospheric plasmas displaying spectra of temperature within or just above physiological ranges allows biological or medical applications of plasmas.

Methods

We have investigated the effects of tissue tolerable plasmas (TTP) on the human pancreatic cancer cell line Colo-357 and PaTu8988T and the murine cell line 6606PDA in vitro (Annexin-V-FITC/DAPI-Assay and propidium iodide DNA staining assay) as well as in the in vivo tumour chorio-allantoic membrane (TUM-CAM) assay using Colo-357.

Results

TTP of 20 seconds (s) induced a mild elevation of an experimental surface temperature of 23.7 degree Celsius up to 26.63+/−0.40 degree Celsius. In vitro TTP significantly (p=0.0003) decreased cell viability showing the strongest effects after 20s TTP. Also, TTP effects increased over time levelling off after 72 hours (30.1+/−4.4% of dead cells (untreated control) versus 78.0+/−9.6% (20s TTP)). However, analyzing these cells for apoptosis 10s TTP revealed the largest proportion of apoptotic cells (34.8+/−7.2%, p=0.0009 versus 12.3+/−6.6%, 20s TTP) suggesting non-apoptotic cell death in the majority of cells after 20s TTP. Using solid Colo-357 tumours in the TUM-CAM model TUNEL-staining showed TTP-induced apoptosis up to a depth of tissue penetration (DETiP) of 48.8+/−12.3μm (20s TTP, p<0.0001). This was mirrored by a significant (p<0.0001) reduction of Ki-67+ proliferating cells (80.9+/−13.2% versus 37.7+/−14.6%, p<0.0001) in the top cell layers as well as typical changes on HE specimens. The bottom cell layers were not affected by TTP.

Conclusions

Our data suggest possible future intra-operative applications of TTP to reduce microscopic residual disease in pancreatic cancer resections. Further promising applications include other malignancies (central liver/lung tumours) as well as synergistic effects combining TTP with chemotherapies. Yet, adaptations of plasma sources as well as of the composition of effective components of TTP are required to optimize their synergistic apoptotic actions.

【 授权许可】

   
2012 Partecke et al.; licensee BioMed Central Ltd.

【 图 表 】

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