Effectiveness of plasma treatment on pancreatic cancer cells

Non-equilibrium atmospheric pressure plasma (NEAPP) has attracted attention in cancer therapy. We explored the indirect effect of NEAPP through plasma-activated medium (PAM) on pancreatic cancer cells in vitro and in vivo. In this study, four pancreatic cancer cell lines were used and the antitumor effects of PAM treatment were evaluated using a cell proliferation assay. To explore functional mechanisms, morphological change and caspase-3/7 activation in cells were also assessed. Furthermore, reactive oxygen species (ROS) generation in cells was examined and N-acetyl cysteine (NAC), an intracellular ROS scavenger, was tested. Finally, the antitumor effect of local injection of PAM was investigated in a mouse xenograft model. We found that PAM treatment had lethal effect on pancreatic cancer cells. Typical morphological findings suggestive of apoptosis such as vacuolization of cell membranes, small and round cells and aggregation of cell nuclei, were observed in the PAM treated cells. Caspase-3/7 activation was detected in accordance with the observed morphological changes. Additionally, ROS uptake was observed in all cell lines tested, while the antitumor effects of PAM were completely inhibited with NAC. In the mouse xenograft model, the calculated tumor volume on day 28 in the PAM treatment group was significantly smaller compared with the control group [28±22 vs. 89±38 (mm3 ± SD), p=0.0031]. These results show that PAM treatment of pancreatic cancer might be a promising therapeutic strategy.

[1]  A. Lowy,et al.  Surgery for pancreatic cancer: recent controversies and current practice. , 2005, Gastroenterology.

[2]  M. Hori,et al.  Nonequilibrium atmospheric pressure plasma with ultrahigh electron density and high performance for glass surface cleaning , 2008 .

[3]  O. Aruoma Free radicals, oxidative stress, and antioxidants in human health and disease , 1998, Journal of the American Oil Chemists' Society.

[4]  M. Ohmura,et al.  CD44 variant regulates redox status in cancer cells by stabilizing the xCT subunit of system xc(-) and thereby promotes tumor growth. , 2011, Cancer cell.

[5]  Woong-Kwon Kim,et al.  DNA damage and mitochondria dysfunction in cell apoptosis induced by nonthermal air plasma , 2010 .

[6]  Jie Liu,et al.  Spheroid body-forming cells in the human gastric cancer cell line MKN-45 possess cancer stem cell properties , 2012, International journal of oncology.

[7]  A. Fridman,et al.  Effects of Non-Thermal Plasma on Mammalian Cells , 2011, PloS one.

[8]  Masaaki Mizuno,et al.  Effectiveness of plasma treatment on gastric cancer cells , 2015, Gastric Cancer.

[9]  N. Sasahira,et al.  Intravenous and Intraperitoneal Paclitaxel with S-1 for Refractory Pancreatic Cancer with Malignant Ascites: an Interim Analysis , 2014, Journal of Gastrointestinal Cancer.

[10]  A. Jemal,et al.  Cancer statistics, 2012 , 2012, CA: a cancer journal for clinicians.

[11]  F Levi,et al.  European cancer mortality predictions for the year 2016 with focus on leukaemias. , 2016, Annals of oncology : official journal of the European Society for Medical Oncology.

[12]  Y. Nishimura,et al.  Microbicidal activities of low frequency atmospheric pressure plasma jets on oral pathogens. , 2011, Dental materials journal.

[13]  P. Brun,et al.  Disinfection of Ocular Cells and Tissues by Atmospheric-Pressure Cold Plasma , 2012, PloS one.

[14]  A. Jemal,et al.  Global Cancer Statistics , 2011 .

[15]  G. Barrera Oxidative Stress and Lipid Peroxidation Products in Cancer Progression and Therapy , 2012, ISRN oncology.

[16]  Ari,et al.  Non-thermal Plasma Induces Apoptosis in Melanoma Cells via Production of Intracellular Reactive Oxygen Species , 2010 .

[17]  M. Nishikawa Reactive oxygen species in tumor metastasis. , 2008, Cancer letters.

[18]  J. Zimmermann,et al.  Focus on Plasma Medicine , 2009 .

[19]  Laura H. Tang,et al.  Survival after Resection of Pancreatic Adenocarcinoma: Results from a Single Institution over Three Decades , 2011, Annals of Surgical Oncology.

[20]  A. le Pape,et al.  Effects of a Non Thermal Plasma Treatment Alone or in Combination with Gemcitabine in a MIA PaCa2-luc Orthotopic Pancreatic Carcinoma Model , 2012, PloS one.

[21]  M. Mizuno,et al.  Plasma-Activated Medium Selectively Kills Glioblastoma Brain Tumor Cells by Down-Regulating a Survival Signaling Molecule, AKT Kinase , 2011 .

[22]  S. Elmore Apoptosis: A Review of Programmed Cell Death , 2007, Toxicologic pathology.

[23]  J. Bahn,et al.  Effects of atmospheric nonthermal plasma on invasion of colorectal cancer cells. , 2010, Applied physics letters.

[24]  Alisa Morss Clyne,et al.  Non-thermal dielectric barrier discharge plasma induces angiogenesis through reactive oxygen species , 2011, 2011 Annual International Conference of the IEEE Engineering in Medicine and Biology Society.

[25]  M. Mizuno,et al.  Effect of Indirect Nonequilibrium Atmospheric Pressure Plasma on Anti-Proliferative Activity against Chronic Chemo-Resistant Ovarian Cancer Cells In Vitro and In Vivo , 2013, PloS one.

[26]  A. Fridman Plasma Chemistry: Frontmatter , 2008 .

[27]  Mari Mino-Kenudson,et al.  Pancreatic ductal adenocarcinoma: long-term survival does not equal cure. , 2012, Surgery.

[28]  K. Oda,et al.  Ratio of CD44 epithelial variant to CD44 hematopoietic variant is a useful prognostic indicator in gastric and colorectal carcinoma. , 1998, Japanese journal of clinical oncology.

[29]  H. Saya,et al.  Inversed relationship between CD44 variant and c-Myc due to oxidative stress-induced canonical Wnt activation. , 2014, Biochemical and biophysical research communications.

[30]  A. Lupu,et al.  Tumoral and Normal Cells Treatment With High-Voltage Pulsed Cold Atmospheric Plasma Jets , 2010, IEEE Transactions on Plasma Science.

[31]  M. Hori,et al.  Selective Killing of Ovarian Cancer Cells through Induction of Apoptosis by a Nonequilibrium Atmospheric Pressure Plasma , 2012 .

[32]  M Keidar,et al.  Cold plasma selectivity and the possibility of a paradigm shift in cancer therapy , 2011, British Journal of Cancer.

[33]  G. Kim,et al.  Biomedical Applications of Low Temperature Atmospheric Pressure Plasmas to Cancerous Cell Treatment and Tooth Bleaching , 2011 .

[34]  Gregory Fridman,et al.  Applied Plasma Medicine , 2008 .

[35]  S. Chevalier,et al.  Mitochondrial DNA damage is sensitive to exogenous H(2)O(2) but independent of cellular ROS production in prostate cancer cells. , 2011, Mutation research.

[36]  Gregory Fridman,et al.  Floating Electrode Dielectric Barrier Discharge Plasma in Air Promoting Apoptotic Behavior in Melanoma Skin Cancer Cell Lines , 2007 .

[37]  K. Song,et al.  Non-Thermal Atmospheric Pressure Plasma Preferentially Induces Apoptosis in p53-Mutated Cancer Cells by Activating ROS Stress-Response Pathways , 2014, PloS one.

[38]  Gregory Fridman,et al.  Blood Coagulation and Living Tissue Sterilization by Floating-Electrode Dielectric Barrier Discharge in Air , 2006 .

[39]  Chul-Ho Kim,et al.  Induction of cell growth arrest by atmospheric non-thermal plasma in colorectal cancer cells. , 2010, Journal of biotechnology.

[40]  T. Yao,et al.  Pathology and prognosis of gastric carcinoma. Findings in 10,000 patients who underwent primary gastrectomy , 1992, Cancer.

[41]  M. Hori,et al.  Laser Scattering Diagnosis of a 60-Hz Non-Equilibrium Atmospheric Pressure Plasma Jet , 2011 .

[42]  B. Györffy,et al.  Biomarkers downstream of RAS: a search for robust transcriptional targets. , 2010, Current cancer drug targets.

[43]  F Levi,et al.  European cancer mortality predictions for the year 2012. , 2012, Annals of oncology : official journal of the European Society for Medical Oncology.

[44]  Eva Stoffels,et al.  Superficial treatment of mammalian cells using plasma needle , 2003 .

[45]  Jue Zhang,et al.  An evaluation of anti-oxidative protection for cells against atmospheric pressure cold plasma treatment , 2012 .

[46]  P. Murawa,et al.  The trifunctional antibody catumaxomab for the treatment of malignant ascites due to epithelial cancer: Results of a prospective randomized phase II/III trial , 2010, International journal of cancer.

[47]  C. Nicco,et al.  Differential modulation of normal and tumor cell proliferation by reactive oxygen species. , 2005, Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie.

[48]  A. Clyne,et al.  Endothelial Cell Proliferation is Enhanced by Low Dose Non-Thermal Plasma Through Fibroblast Growth Factor-2 Release , 2010, Annals of Biomedical Engineering.

[49]  J. Sobilo,et al.  Antitumor Effect of Plasma Treatment on U87 Glioma Xenografts: Preliminary Results , 2010 .

[50]  D. Graham,et al.  The Role of CD44 in the Pathogenesis, Diagnosis, and Therapy of Gastric Cancer , 2011, Gut and liver.

[51]  C. Figdor,et al.  De-novo expression of CD44 and survival in gastric cancer , 1993, The Lancet.

[52]  H. Nagawa,et al.  Phase II study of weekly intravenous and intraperitoneal paclitaxel combined with S-1 for advanced gastric cancer with peritoneal metastasis. , 2009, Journal of clinical oncology : official journal of the American Society of Clinical Oncology.

[53]  D. Levison,et al.  Pathological prognostic factors in the second British Stomach Cancer Group trial of adjuvant therapy in resectable gastric cancer. , 1995, British Journal of Cancer.

[54]  M. Imamura,et al.  Evaluation of extra- and intracellular OH radical generation, cancer cell injury, and apoptosis induced by a non-thermal atmospheric-pressure plasma jet , 2013 .

[55]  Takehiko Sato,et al.  A key inactivation factor of HeLa cell viability by a plasma flow , 2011 .

[56]  R. Burger,et al.  Intraperitoneal cisplatin and paclitaxel in ovarian cancer. , 2006, The New England journal of medicine.

[57]  S. Yamada,et al.  Overexpression of L1CAM is Associated with Tumor Progression and Prognosis via ERK Signaling in Gastric Cancer , 2014, Annals of Surgical Oncology.

[58]  D. Klimstra,et al.  Long-term survival after curative resection for pancreatic ductal adenocarcinoma. Clinicopathologic analysis of 5-year survivors. , 1996, Annals of surgery.

[59]  J. Donohue,et al.  Long-Term Survival After Pancreatoduodenectomy for Pancreatic Adenocarcinoma: Is Cure Possible? , 2008, Annals of surgery.

[60]  S. Saglam,et al.  Long-term survival after curative resection for pancreatic ductal adenocarcinoma. , 2004, Journal of clinical oncology : official journal of the American Society of Clinical Oncology.

[61]  Retraction Note to: Non-thermal Plasma Induces Apoptosis in Melanoma Cells via Production of Intracellular Reactive Oxygen Species , 2011, Annals of Biomedical Engineering.

[62]  J. Furuse,et al.  The Hepatobiliary and Pancreatic Oncology (HBPO) Group of the Japan Clinical Oncology Group (JCOG): history and future direction. , 2013, Japanese journal of clinical oncology.