Examination of immunomodulatory and cytotoxic effects of aqueous exstract of white mistletoe (Viscum album L.) in vitro and in patients treated with anthracycline chemotherapeutics

Uspešnost antitumorske terapije je ograničena pojavom tumorske rezistencije na lekove putem molekularnih mehanizama koji i dalje ostaju nerazjašnjeni. Kako bi se smanjili neželjeni efekti i poboljšala efikasnost standardnih hemioterapija i radioterapija, počeli su da se primenjuju mnogi komplementarni ili alternativni medikamenti. Bela imela (lat. Viscum album L) se tradicionalno koristi u lečenju pojedinih vrsta tumora. Terapijska korist od primene bele imele kada se koristi zajedno sa hemioterapijom ili radioterapijom, odnosno operacijom doprinosi ukupnom poboljšanju kvaliteta života kod pacijenata obolelih od tumora. Dosadašnja istraživanja na ćelijama u kulturi i na životinjskim modelima, su pokazala da potencijalni mehanizmi delovanja sastojaka bele imele uključuju antitumorsko, antiinflamatorno i imunomodulatorno dejstvo. Uprkos intezivnim ispitivanjima, mehanizam antitumorskog dejstva ekstrakta bele imele je još uvek u velikom obimu nepoznat. Takođe, malo je i naučnih podataka o mehanizmu interakcije bioaktivnih komponenti bele imele i drugih antitumorskih citotoksičnih lekova, koji bi pružili zasnovanost daljem kliničkom ispitivanju. Cilj ove disertacije je utvrđivanje imunomodulatornog dejstva ekstrakta bele imele na ćelije imunskog sistema u in vitro i in vivo uslovima, ispitivanje citotoksičnog efekta ekstrakta bele imele in vitro i molekularnih mehanizma interakcije bioaktivnih komponenti ekstrakta bele imele sa citotoksičnim agensom iz grupe antraciklina na modelu tumorskih ćelija u kulturi. Rezultati dobijeni u in vitro istraživanjima su pokazali citotoksični i antiproliferativni efekat ekstrakta bele imele, putem indukcije apoptoze tumorskih ćelija unutrašnjim mitohondrijskim putem i indukcijom zastoja u G0/G1 fazi ćelijskog ciklusa. Kombinovani tretman tumorskih ćelija subterapeutskom koncentracijom doksorubicina i ekstraktom bele imele dovodi do sinergističkog inhibitornog efekta...Success of anticancer treatment has been impeded by resistance to anti-tumor therapy via molecular mechanisms that still remain elusive. To reduce side effects and improve the effectiveness of standard chemoradiation therapy, many complementary or alternative medicines (CAM) have been investigated. Mistletoe (Viscum album L) has been long used as CAM supporting cancer therapy. Therapeutic benefit of mistletoe preparations when utilized along with surgery, chemotherapy or radiotherapy contributes to the overall improvement in the quality of life in cancer patients. Several lines of evidence have revealed that mistletoe preparations in vitro and in vivo exert an anti-cancer, anti-inflamatory and immunomodulatory effects. Despite extensive experimental analyses of their biological properties, many questions regarding the precise mode of action of mistletoe still remain elusive. Also, there is only little information about interactions between mistletoe and anticancer drugs, which would provide validity for further clinical investigations. The aims of this study are investigation of immunomodulatory effects of mistletoe extracts on immune cells in vitro and in vivo, examination of cytotoxic effect in vitro, as well as molecular mechanisms underlying interactions between mistletoe and anticancer drug from the antraciklenes family on the tumor cells in culture. Our results from in vivo study study have shown cytotoxic and antiproliferative effect of mistletoe extract via induction of apoptosis of cancer cells through intrinsic mitochondrial pathway and induction of G0/G1 arrest. Also, combined treatment of cancer cells with sub-therapeutic concentration of doxorubicine and mistletoe extract led to synergistic inhibitory effect. Increased K562 and MCF7 cells apoptotic death during cotreatment was associated with reduced G2/M accumulation during doxorubicine treatment..

Additional Therapy with a Mistletoe Product during Adjuvant Chemotherapy of Breast Cancer Patients Improves Quality of Life: An Open Randomized Clinical Pilot Trial

Background. Breast cancer patients receiving adjuvant chemotherapy often experience a loss of quality of life. Moreover chemotherapy may induce neutropenia. Patients report a better quality of life when additionally treated with mistletoe products during chemotherapy. Methods. In this prospective randomized open-label pilot study 95 patients were randomized into three groups. All patients were treated with an adjuvant chemotherapy. The primary objective of the study was quality of life, the secondary objective was neutropenia. Here we report the comparison of HxA (n = 34) versus untreated control (n = 31). Results. In the explorative analysis ten of 15 scores of the EORTC QLQ-C30 showed a better quality of life in the HxA group compared to the control group ( < 0.001 to = 0.038 in Dunnett-T3 test). The difference was clinically relevant (difference of at least 5 points, range 5.4-12.2) in eight of the ten scores. Neutropenia occurred in 7/34 HxA patients and in 8/31 control patients (P = 0.628). Conclusions. This pilot study showed an improvement of quality of life by treating breast cancer patients with HxA additionally to CAF. Although the open design may be a limitation, the findings show the feasibility of a confirmatory study using the methods described here

Quality of Life and Neutropenia in Patients with Early Stage Breast Cancer: A Randomized Pilot Study Comparing Additional Treatment with Mistletoe Extract to Chemotherapy Alone

Background: Chemotherapy for breast cancer often deteriorates quality of life, augments fatigue, and induces neutropenia. Mistletoe preparations are frequently used by cancer patients in Central Europe. Physicians have reported better quality of life in breast cancer patients additionally treated with mistletoe preparations during chemotherapy. Mistletoe preparations also have immunostimulant properties and might therefore have protective effects against chemotherapy-induced neutropenia.Patients and Methods: We conducted a prospective randomized open label pilot study with 95 patients randomized into three groups. Two groups received Iscador® M special (IMS) or a different mistletoe preparation, respectively, additionally to chemotherapy with six cycles of cyclophosphamide, adriamycin, and 5-fluoro-uracil (CAF). A control group received CAF with no additional therapy. Here we report the comparison IMS (n = 30) vs. control (n = 31). Quality of life including fatigue was assessed with the European Organization for Research and Treatment of Cancer Quality of Life Questionnaire (EORTC-QLQ-C30). Neutropenia was defined as neutrophil counts <1,000/µl and assessed at baseline and one day before each CAF cycle.Results: In the descriptive analysis all 15 scores of the EORTC-QLQ-C30 showed better quality of life in the IMS group compared to the control group. In 12 scores the differences were significant (p < 0.02) and nine scores showed a clinically relevant and significant difference of at least 5 points. Neutropenia occurred in 3/30 IMS patients and in 8/31 control patients (p = 0.182).Conclusions: This pilot study showed an improvement of quality of life by treating breast cancer patients with IMS additionally to CAF. CAF-induced neutropenia showed a trend to lower frequency in the IMS group

Iscador Qu inhibits doxorubicin-induced senescence of MCF7 cells (vol 7, 3763, 2017)

A correction to this article has been published and is linked from the HTML and PDF versions of this paper. The error has been fixed in the paper

Cell death in irradiated prostate cancer cells assessed by flow cytometry

Despite the significant advances in cancer chemotherapy, radiotherapy still remains a method of choice for treatment of metastatic human prostate cancer. This study presents quantitative analysis of 60Co gamma-radiation effects on cell growth and cell death of metastatic human prostate cancer PC-3 cell line, performed in time (24-72h) and dose (2-20 Gy) dependent manner. The irradiated PC-3 cells were mostly dying by necrosis at late time intervals (72h), while apoptotic cell death was negligible. The EC50 or 50% of cytotoxicity was not achieved within the radiation doses used (2-20 Gy), but significant cell growth inhibition with IC50 of 10.4 Gy was observed. It is concluded that the increase in the radiation dose may have an important cytostatic effect, but for the complete eradication of metastatic prostate cancer novel cytotoxic drugs and radiosensitizers should be introduced as adjuvant.Uprkos značajnom napretku u hemoterapiji kancera, radioterapija ostaje metod izbora u tretmanu metastaziranog kancera prostate. Ovaj rad predstavlja kvantitativnu analizu efekata 60Co gama zračenja na ćelijski rasti i ćelijsku smrt PC-3 ćelijske linije humanog kancera prostate, pri čemu je praćena vremenska (2-72h) i dozna zavisnost (2-20 Gy). Ozračene PC-3 ćelije su uglavnom umirale nekrozom u kasnijem vremenskom intervalu (72h), dok je apoptoza bila zanemarljiva. Vrednost EC50 odnosno 50% citotoksičnosti nije dostignuta primenjenim dozama, ali je ustanovljena značajna inhibicija ćelijskog rasta, sa vrednošću IC50 od 10.4 Gy. Zaključeno je da povećanje doze može imati značajan citostatički efekat ali da je za kompletno odstranjivanje metastaziranog kancera prostate neophodno uvođenje novih citotoksičnih agenasa ili radiosenzitera kao adjuvanata.nul

Efekti ekstrakta bele imele na markere aktivacije i agregacije trombocita

BACKGROUND: Viscum album preparations are extensively used as complementary therapy in cancer and are shown to exert antitumor activities which involve the cytotoxic properties, induction of apoptosis, inhibition of angiogenesis and several other immunomodulatory mechanisms. AIM: The aim of this study was to investigate the eft fects of mistletoe extract on platelet as well as monocyte functions, as an important factors in immunomodulation of cancers metas tatic potencial and angiogenesis in tumors. METHODS: The effect of different concentrations of mistletoe extract on agonist-induced platelet activation markers and their aggregation with leukocytes was examined in the blood of healthy subjects ( n = 6 ) using flow cytometry. Effects on LPS -induced activation markers was det ermined in the blood of healthy subj ects as well as on THP- 1 cell line using an ELISA essays and flow cytometry. RESULTS: Mistletoe extract significantly inhibited agonist induced P selectin expression and platelet-monocytes aggregation. Additionally, mistletoe extract exerts anti-tumor effect through the stimulation of TNF-a production in LPS induced monocytes activation. CONCLUSION: Obtained data demonstrate that mistletoe extract was effective in modulating platelet and monocyte functions, as a part of pleiotropic anticancer effect.UVOD: Preparati biljke Viscum album se intenzivno koriste kao komplementarna terapija u lecenju kancera. Mehanizmi antitumorskog delovanja, potvrđeni in vitro, ukljucuju citotoksicno delovanje, indukciju apoptoze, inhibiciju angiogeneze, imunomodulatorno delovanje. CILJ: Cilj ovog istraživanja je ispitivanje uticaja ekstrakta bele imele na funkciju trombocita i monocita kao važnih faktora u imunomodulaciji kancerskog procesa, metastatskom potencijalu i tumorskoj angiogenezi. METODE: Uticaj razlicitih koncentracija ekstrakta bele imele na markere agonistom indukovane aktivacije trombocita i njihove agregacije sa leukocitima ispitivan je u krvi zdravih ispitanika (n=6) primenom protocne citometrije. Uticaj na markere LPS indukovane aktivacije određivan je u krvi ispitanika i kulturi THP-1 celija korišceenjem ELISA eseja i protocne citometrije. REZULTATI: Ekstrakt imele znacajno inhibira ekspresiju P-selektina i trombocitno-monocitnu agregaciju. Pokazana stimulacija produkcije TNF-a u LPS-om aktiviranim monocitima dodatno doprinosi antitumorskom potencijalu. ZAKLJUČAK: Dobijeni rezultati potvrđuju potenci jal ekstrakta imele da modulira trombocitnu i monocitnu funkciju, kao deo pleotropnog antitumorskog delovanja

Examination of immunomodulatory and cytotoxic effects of aqueous exstract of white mistletoe (Viscum album L.) in vitro and in patients treated with anthracycline chemotherapeutics

Uspešnost antitumorske terapije je ograničena pojavom tumorske rezistencije na lekove putem molekularnih mehanizama koji i dalje ostaju nerazjašnjeni. Kako bi se smanjili neželjeni efekti i poboljšala efikasnost standardnih hemioterapija i radioterapija, počeli su da se primenjuju mnogi komplementarni ili alternativni medikamenti. Bela imela (lat. Viscum album L) se tradicionalno koristi u lečenju pojedinih vrsta tumora. Terapijska korist od primene bele imele kada se koristi zajedno sa hemioterapijom ili radioterapijom, odnosno operacijom doprinosi ukupnom poboljšanju kvaliteta života kod pacijenata obolelih od tumora. Dosadašnja istraživanja na ćelijama u kulturi i na životinjskim modelima, su pokazala da potencijalni mehanizmi delovanja sastojaka bele imele uključuju antitumorsko, antiinflamatorno i imunomodulatorno dejstvo. Uprkos intezivnim ispitivanjima, mehanizam antitumorskog dejstva ekstrakta bele imele je još uvek u velikom obimu nepoznat. Takođe, malo je i naučnih podataka o mehanizmu interakcije bioaktivnih komponenti bele imele i drugih antitumorskih citotoksičnih lekova, koji bi pružili zasnovanost daljem kliničkom ispitivanju. Cilj ove disertacije je utvrđivanje imunomodulatornog dejstva ekstrakta bele imele na ćelije imunskog sistema u in vitro i in vivo uslovima, ispitivanje citotoksičnog efekta ekstrakta bele imele in vitro i molekularnih mehanizma interakcije bioaktivnih komponenti ekstrakta bele imele sa citotoksičnim agensom iz grupe antraciklina na modelu tumorskih ćelija u kulturi. Rezultati dobijeni u in vitro istraživanjima su pokazali citotoksični i antiproliferativni efekat ekstrakta bele imele, putem indukcije apoptoze tumorskih ćelija unutrašnjim mitohondrijskim putem i indukcijom zastoja u G0/G1 fazi ćelijskog ciklusa. Kombinovani tretman tumorskih ćelija subterapeutskom koncentracijom doksorubicina i ekstraktom bele imele dovodi do sinergističkog inhibitornog efekta...Success of anticancer treatment has been impeded by resistance to anti-tumor therapy via molecular mechanisms that still remain elusive. To reduce side effects and improve the effectiveness of standard chemoradiation therapy, many complementary or alternative medicines (CAM) have been investigated. Mistletoe (Viscum album L) has been long used as CAM supporting cancer therapy. Therapeutic benefit of mistletoe preparations when utilized along with surgery, chemotherapy or radiotherapy contributes to the overall improvement in the quality of life in cancer patients. Several lines of evidence have revealed that mistletoe preparations in vitro and in vivo exert an anti-cancer, anti-inflamatory and immunomodulatory effects. Despite extensive experimental analyses of their biological properties, many questions regarding the precise mode of action of mistletoe still remain elusive. Also, there is only little information about interactions between mistletoe and anticancer drugs, which would provide validity for further clinical investigations. The aims of this study are investigation of immunomodulatory effects of mistletoe extracts on immune cells in vitro and in vivo, examination of cytotoxic effect in vitro, as well as molecular mechanisms underlying interactions between mistletoe and anticancer drug from the antraciklenes family on the tumor cells in culture. Our results from in vivo study study have shown cytotoxic and antiproliferative effect of mistletoe extract via induction of apoptosis of cancer cells through intrinsic mitochondrial pathway and induction of G0/G1 arrest. Also, combined treatment of cancer cells with sub-therapeutic concentration of doxorubicine and mistletoe extract led to synergistic inhibitory effect. Increased K562 and MCF7 cells apoptotic death during cotreatment was associated with reduced G2/M accumulation during doxorubicine treatment..

Additional Therapy with a Mistletoe Product during Adjuvant Chemotherapy of Breast Cancer Patients Improves Quality of Life: An Open Randomized Clinical Pilot Trial

Background. Breast cancer patients receiving adjuvant chemotherapy often experience a loss of quality of life. Moreover chemotherapy may induce neutropenia. Patients report a better quality of life when additionally treated with mistletoe products during chemotherapy. Methods. In this prospective randomized open-label pilot study 95 patients were randomized into three groups. All patients were treated with an adjuvant chemotherapy. The primary objective of the study was quality of life, the secondary objective was neutropenia. Here we report the comparison of HxA (n = 34) versus untreated control (n = 31). Results. In the explorative analysis ten of 15 scores of the EORTC QLQ-C30 showed a better quality of life in the HxA group compared to the control group (P<0.001 to P=0.038 in Dunnett-T3 test). The difference was clinically relevant (difference of at least 5 points, range 5.4–12.2) in eight of the ten scores. Neutropenia occurred in 7/34 HxA patients and in 8/31 control patients (P = 0.628). Conclusions. This pilot study showed an improvement of quality of life by treating breast cancer patients with HxA additionally to CAF. Although the open design may be a limitation, the findings show the feasibility of a confirmatory study using the methods described here

Cell death in irradiated prostate cancer cells assessed by flow cytometry

Despite the significant advances in cancer chemotherapy, radiotherapy still remains a method of choice for treatment of metastatic human prostate cancer. This study presents quantitative analysis of 60Co gamma-radiation effects on cell growth and cell death of metastatic human prostate cancer PC-3 cell line, performed in time (24-72h) and dose (2-20 Gy) dependent manner. The irradiated PC-3 cells were mostly dying by necrosis at late time intervals (72h), while apoptotic cell death was negligible. The EC50 or 50% of cytotoxicity was not achieved within the radiation doses used (2-20 Gy), but significant cell growth inhibition with IC50 of 10.4 Gy was observed. It is concluded that the increase in the radiation dose may have an important cytostatic effect, but for the complete eradication of metastatic prostate cancer novel cytotoxic drugs and radiosensitizers should be introduced as adjuvant

Radiation-induced effects in PC-3 and DU-145 human prostate cancer cells

Background: Prostate cancer is the first as an incidence and the second as a cause of the oncologic mortality in the male population. There is a broad range of possibilities in the prostate cancer therapy. However, there is also much controversy on the most appropriate treatment in the various stages of the disease. Advanced disease is mostly treated by radiation therapy, sometimes in combination with hormone or chemotherapy. Irradiation induces damage of cell biomolecules, which can lead to the arrest in cell division, or to apoptotic or necrotic cell death. The aim of this study was to determine the dose dependence of radiation-induced cell death in two human prostate cancer cell lines, and to define the form of death of these cells. Methods: Human prostate cancer cell lines PC-3 and DU-145 were irradiated with 2 - 30 Gy from 60 Co g-source, at the dose rate of 20 Gy/h. The effect of irradiation on cell viability, morphology and DNA structure were followed 24 - 72 hours after treatment. Cells were analyzed by trypan blue exclusion assay, flow cytometry and DNA gel electrophoresis. Simultaneous staining of cells with Annexin V-FITC and propidium iodide enabled distinction of early apoptosis from late apoptosis and/or necrosis. Results: The results of trypan blue staining indicated that radiation-induced cell death was both time and dose dependent process. According to flow-cytometry and DNA fragmentation assay, necrosis was the prevailing form of the radiation-induced cell death in both PC-3 and DU-145 cells. The apoptosis occurred in insignificant number of cells, probably due to the mutant p53 gene present in both cell lines. The cell necrosis was dose dependent and was most pronounced 72 hours post treatment. Conclusion The prevailing form of radiation-induced PC-3 and DU-145 cell death is necrosis. Both PC-3 and DU-145 are rather radioresistant cell lines, as the dose necessary to induce 50% decrease in viable cell number is about 10 Gy.nul