Cold Atmospheric Plasma Does Not Affect Stellate Cells Phenotype in Pancreatic Cancer Tissue in Ovo

Abstract

Pancreatic ductal adenocarcinoma (PDAC) is a challenging neoplastic disease, mainly due to the development of resistance to radio- and chemotherapy. Cold atmospheric plasma (CAP) is an alternative technology that can eliminate cancer cells through oxidative damage, as shown in vitro, in ovo, and in vivo. However, how CAP affects the pancreatic stellate cells (PSCs), key players in the invasion and metastasis of PDAC, is poorly understood. This study aims to determine the effect of an anti-PDAC CAP treatment on PSCs tissue developed in ovo using mono- and co-cultures of RLT-PSC (PSCs) and Mia PaCa-2 cells (PDAC). We measured tissue reduction upon CAP treatment and mRNA expression of PSC activation markers and extracellular matrix (ECM) remodelling factors via qRT-PCR. Protein expression of selected markers was confirmed via immunohistochemistry. CAP inhibited growth in Mia PaCa-2 and co-cultured tissue, but its effectiveness was reduced in the latter, which correlates with reduced ki67 levels. CAP did not alter the mRNA expression of PSC activation and ECM remodelling markers. No changes in MMP2 and MMP9 expression were observed in RLT-PSCs, but small changes were observed in Mia PaCa-2 cells. Our findings support the ability of CAP to eliminate PDAC cells, without altering the PSCs.

Conductive Gas Plasma Treatment Augments Tumor Toxicity of Ringer’s Lactate Solutions in a Model of Peritoneal Carcinomatosis

“…In addition to immune cell suppression, desmoplastic reactions of tumor resident stromal fibroblasts can occur, e.g., pancreatic stellate cells (PSC) in the context of pancreatic ductal carcinoma, which are a major cause of therapy resistance and poor patient outcome. Direct and indirect plasma treatment regimens have recently been shown to counteract PSC activation and extracellular matrix remodeling [ 46 , 59 , 60 , 61 ], as well as exuberant proliferation and activity of stromal fibroblasts after peritoneal surgery and prolonged inflammation [ 62 ], which translates to conditions in the TME that might be additionally beneficial after cytoreductive surgery in PC.…”

The effect of local non‐thermal plasma therapy on the cancer‐immunity cycle in a melanoma mouse model

“…Immune cells in the NTP‐treated mice also had higher cytotoxic capacity in the tumor, as indicated by increased GrB expression in both CD8 + T cells and NK cells. The effect of NTP on other cell types within the tumor has also been previously reported (e.g., cancer‐associated fibroblasts, 50 tumor‐associated macrophages 51 ) and will be investigated in future studies. Furthermore, it is of great interest to determine the localization of recruited immune cells in the tumor microenvironment with respect to the site of NTP treatment.…”

Cold atmospheric plasma: Novel opportunities for tumor microenvironment targeting

“…Since the first discovery on the anti‐cancer efficacy of CAP in 2007, consecutive efforts have been devoted to investigate its onco‐therapeutic impacts in varied types of cancers with demonstrated efficacies already been proven in, for example, triple negative breast cancers, 201 bladder cancers, 202 prostate cancers, 203 melanomas, 204 and pancreatic cancers. 205 Differential cell death events can be triggered by CAP in a dose‐dependent manner 206 that include, for example, cell cycle arrest, 203 autophagy, 207 apoptosis, 201 ferroptosis, 208 immunogenic cell death (ICD), 209 and necrotic cell death. 210 It has also been proposed that CAP can modulate the immunogenic response 211 and drug sensitivity 212 of cancer cells, halt cancer invasion and metastasis, 202 and rewire the metabolic reprogramming of malignant cells, 213 among others.…”

“…Cold atmospheric plasma is composed of varied reactive oxygen and nitrogen species (RONS) including short‐lived species such as hydroxyl radical (OH·), singlet oxygen (O), superoxide (O 2− ), and nitric oxide (NO·), and long‐lived species such as hydrogen peroxide (H 2 O 2 ), ozone (O 3 ), anionic (OONO − ), and protonated (ONOOH) forms of peroxynitrite. Since the first discovery on the anti‐cancer efficacy of CAP in 2007, consecutive efforts have been devoted to investigate its onco‐therapeutic impacts in varied types of cancers with demonstrated efficacies already been proven in, for example, triple negative breast cancers, 201 bladder cancers, 202 prostate cancers, 203 melanomas, 204 and pancreatic cancers 205 . Differential cell death events can be triggered by CAP in a dose‐dependent manner 206 that include, for example, cell cycle arrest, 203 autophagy, 207 apoptosis, 201 ferroptosis, 208 immunogenic cell death (ICD), 209 and necrotic cell death 210 .…”