ETM study of electroporation influence on cell morphology in human malignant melanoma and human primary gingival fibroblast cells

Skołucka, Nina; Daczewska, Małgorzata; Saczko, Jolanta; Chwiłkowska, Agnieszka; Choromańska, Anna; Kotulska, Małgorzata; Kamińska, Iwona; Kulbacka, Julita

doi:10.1016/s2221-1691(11)60003-8

Cited by 13 publications

(10 citation statements)

References 15 publications

(18 reference statements)

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“…However, recent mathematical calculations have shown that such pulses can actually have effects on internal membranes 49 . This theoretical model has been confirmed in vitro by our laboratory (unpublished data) and by others, although a 2-fold higher field amplitude was used in this latter study 50 . Hence, the microsecond electric pulses used in our study and in ECT might directly provoke CRT translocation to the cell membrane.…”

Section: Discussionsupporting

confidence: 77%

Electrochemotherapy with bleomycin induces hallmarks of immunogenic cell death in murine colon cancer cells

Calvet

Famin

André³

et al. 2014

OncoImmunology

171

173

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Electrochemotherapy (ECT) is a local cancer treatment that has been used over the course of more than 2 decades for the removal of cutaneous and subcutaneous tumors. Several lines of evidence support the premise that the immune system is an important factor underlying anticancer treatment efficacy, potentially including patient responses to ECT. The concept of immunogenic cell death (ICD) arose a few years ago, stating that some cancer treatments generate danger-associated molecular patterns (DAMPs) that trigger an adaptive immune response against tumors. Hence, dying cancer cells behave as a therapeutic vaccine, eliciting a cytotoxic immune response against surviving malignant cells. In our study, we sought to evaluate the ability of ECT to generate cancer cell death encompassing the immunostimulatory characteristics of ICD. To this end, we assayed CT26 murine colon cancer cells in vitro in response to either electric pulses (EPs) application only or in combination with the anticancer drug bleomycin (that is ECT) by quantification of calreticulin (CRT) membrane externalization, as well as the liberation of adenosine triphosphate (ATP) and high mobility group box 1 (HMGB1) protein. We show here that cell permeabilizing yet non-lethal electric pulses induce CRT exposure on the cell surface of EP-only treated cancer cells, as well as ATP release. However, the association of electric pulses along with the chemotherapeutic agent bleomycin was mandatory for HMGB1 release coincident with regimen-induced cell death. These data obtained in vitro were then substantiated by vaccination protocols performed in immunocompetent mice, showing that the injection of dying ECT-treated cells elicits an antitumor immune response that prevents the growth of a subsequent administration of viable cancer cells. We also confirmed previous results showing ECT treatment is much more efficient in immunocompetent animals than in immunodeficient ones, causing complete regressions in the former but not in the latter. This supports a central role for immunity in this beneficial outcome. In conclusion, we show that ECT not only possesses an intrinsic cytotoxic property toward cancer cells but also generates a systemic anticancer immune response via the activation of ICD. Hence, ECT may represent an interesting approach to treat solid tumors while preventing recurrence and metastasis, possibly in combination with immunostimulating agents.

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Section: Discussionsupporting

confidence: 77%

Electrochemotherapy with bleomycin induces hallmarks of immunogenic cell death in murine colon cancer cells

Calvet

Famin

André³

et al. 2014

OncoImmunology

171

173

View full text Add to dashboard Cite

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“…As expected, immature cardiomyocytes show significantly lower viability threshold value of the pulse intensities than cells of other tissues treated by the same protocol (e.g. Saczko et al 2010;Skolucka et al 2011). The effect of EP above this pulse intensity can also be observed on phase contrast images of cells, obtained directly after EP.…”

Section: Discussionsupporting

confidence: 72%

Electroporation-induced changes in normal immature rat myoblasts (H9C2)

Kamińska¹,

Kotulska²,

Stecka³

et al. 2012

gpb

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Application of a high electric field causes an electric shock to the heart. This is utilized in defibrillation to reestablish normal contraction rhythms during dangerous arrhythmias or in cardiac arrest. If shock-induced transmembrane potentials are large enough, they can cause tissue destruction due to irreversible electroporation (EP). Also electrochemotherapy of nearby tissues may have an adverse effect on the heart. Herein, we present experimental data on effects of electroporation in culture of cardiac cells (H9C2). The electric field was applied in short pulses of 25-3250 V/cm, 50 µs each. The viability of cells was tested by MTT assay after 24 hours. For detection of DNA fragmentation, associated with apoptosis, alkaline and neutral comet assays were performed after EP. Additionally phase contrast images of cells obtained directly after EP were analyzed. Although cell images indicated disruption of cell membranes after EP with high intensities, only a few percent of apoptotic cells and no necrotic effects in the cell nucleus could be observed in comet assay tests performed 2 hours post EP. MTT viability test showed that pulse intensities above 375 V/cm are destructive for myocytes viability.

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“…The electro-photodynamic therapy applied to lung cancer cells (A549) revealed a significant increase of the cytotoxic effect on cells when compared to HpD-PDT alone, even at drug doses significantly lower than typical levels. The results also demonstrated that the drug was delivered within minutes, instead of hours as in standard PDT (Skolucka et al 2011). Labanauskiene et al (2009 examined Chinese hamster lung fibroblast cell line (DC-3F) (2007) and murine hepatoma MH22A cells.…”

Section: Discussionmentioning

confidence: 78%

“…The Epon blocks were cut on the ultramicrotome (Ultracut E, Reihert, Germany). Ultrathin sections were contrasted with uranyl acetate and lead citrate according to the method described by Skolucka et al (2011) and examined with a TEM Zeiss EM 900 (Carl Zeiss, Oberkochen, Germany).…”

Section: Transmission Electron Microscope (Tem)mentioning

confidence: 99%

Cellular stress induced by photodynamic reaction with CoTPPS and MnTMPyPCl5 in combination with electroporation in human colon adenocarcinoma cell lines (LoVo and LoVoDX)

Kulbacka¹,

Kotulska

Rembiałkowska³

et al. 2013

Cell Stress and Chaperones

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Two porphyrins, CoTPPS and MnTMPyPCl 5 , were tested for their photodynamic activity and potential novel use in a therapy of human cancers. We investigated an effect of photodynamic reaction (PDR), electroporation (EP) and their combination (electro-photodynamic reaction [EP-PDR]) on human colon adenocarcinoma cell lines (LoVo and resistant to doxorubicin LoVoDX), human breast adenocarcinoma (wild type MCF-7/WT and resistant to doxorubicin MCF-7/DOX), and human melanoma (Me45). The efficiency of macromolecules transport was examined with cytofluorymetry by assessing the degree of propidium iodide (PI) penetration. Additionally, cellular ultrastructure after EP was evaluated. We determined cyto-and photo-cytotoxic effect on the cells viability (MTT assay) after standard PDR and PDR combined with EP. Intracellular distribution and mitochondrial colocalization of both porphyrins was also performed. The experiments proved that both complexes exhibit desirable photodynamic properties on LoVo LoVoDX cells, and EP effectively supports photodynamic method in this type of cancer. The application of EP provided shorter time of incubation (only 10 min) and enhanced effect of applied therapy. The porphyrins did not affect the MCF-7 and Me45 cell lines.

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ETM study of electroporation influence on cell morphology in human malignant melanoma and human primary gingival fibroblast cells

Abstract: Asian Pac J Trop Biomed 2011; 1(2): 94-98

Cited by 13 publications

References 15 publications

Electrochemotherapy with bleomycin induces hallmarks of immunogenic cell death in murine colon cancer cells

Electrochemotherapy with bleomycin induces hallmarks of immunogenic cell death in murine colon cancer cells

Electroporation-induced changes in normal immature rat myoblasts (H9C2)

Cellular stress induced by photodynamic reaction with CoTPPS and MnTMPyPCl5 in combination with electroporation in human colon adenocarcinoma cell lines (LoVo and LoVoDX)

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