Effect of ethanolic extract of lady's mantle (Alchemilla vulgaris agg.) on malignant cell phenotype and tumor microenvironment in syngenic murine melanoma and breast cancer models

[S. A. Jelača]

137 str.

Biologija tumora - Eksperimentalna onkologija / Tumor biology - Experimental oncology

Virak (Alchemilla vulgaris agg.) je biljka sa dugom istorijom upotrebe u narodnoj medicini koja se koristi u lečenju različitih bolesti. U ovoj studiji ispitan je uticaj etanolnog ekstrakta A. vulgaris na rast ćelijskih linija melanoma i dojke različitog porekla in vitro dok su efekti in vivo praćeni na mišjim modelima melanoma (B16F1 i B16F10) i tumora dojke (4T1). Takođe, analiziran je i uticaj na lokalni i sistemski imunski odgovor u modelima B16F1 i 4T1. Pokazano je da ekstrakt ostvaruje snažan antitumorski efekat na svim ispitivanim linijama. Tretman ekstraktom je doveo do inhibicije proliferacije i indukcije smrti autofagijom u B16F1 ćelijama, dok je u B16F10 klonu suprimirana deoba praćena apoptozom nezavisnom od aktivacije kaspaza. Slično, 4T1 ćelije su pod tretmanom podlegle programiranoj ćelijskoj smrti tipa I i II dok su im proliferativni i klonogeni potencijal, adhezivnost, migratornost i invazivnost smanjeni kroz inhibiciju integrinskog signalnog puta. Delotvornost tretmana je potvrđena in vivo, sa značajnim smanjenjem finalnog volumena tumora u sva tri modela. Pored direktnog uticaja na tumorske ćelije u modelima B16F1 i 4T1 in vivo, ekstrakt stimuliše sistemski i lokalni antitumorski imunski odgovor. Imunosupresivno okruženje je potisnuto maturacijom dendritskih ćelija sa poboljšanom sposobnošću da prezentuju antigene i doprinesu aktivaciji CD8+ T limfocita u oba modela. Dodatno, u modelu B16F1, tretman je povećao procenat Th-1 ćelija koje produkuju proinflamatorne citokine i smanjio prisustvo regulatornih T ćelija i proizvodnju antiinflamatornih citokina. Svi navedeni efekti su praćeni odsustvom toksičnosti. Rezultati ove studije predstavljaju prve naučne podatke o višestrukim antitumorskim potencijalima A. vulgaris ekstrakta in vivo.

Alchemilla vulgaris agg. is a plant with a long history of use in folk medicine for the treatment of various diseases. This study investigated the effect of the ethanolic extract of A. vulgaris on melanoma and breast cancer cell lines in vitro, and its effect in murine models of melanoma (B16F1 and B16F10) and breast tumor (4T1) in vivo. Moreover, the impact on local and systemic immune response was analyzed in the B16F1 and 4T1 models. The results showed that the extract exerted a strong antitumor effect on all tested cells. In B16F1 cells, the treatment inhibited proliferation and induced autophagic cell death, while in B16F10 counterpart, the treatment led to caspase-independent apoptosis along with inhibition of proliferation. Similarly, 4T1 cells underwent type I and II programmed cell death under treatment, with reduced proliferative and clonogenic potential, adhesion, migration, and invasion due to inhibition of the integrin signaling pathway. The efficacy of the treatment was also confirmed in vivo, with a significant reduction in final tumor volume in all three models. Besides direct effects on tumor cells in B16F1 and 4T1 models, the extract significantly promoted systemic and local antitumor immune response. In both models, the immunosuppressive environment was alleviated by the maturation of dendritic cells, which showed an enhanced ability to present antigens and activate CD8+ T cells. Also, in the B16F1 model, the extract increased the proportion of Th-1 cells producing pro-inflammatory cytokines, while it reduced the presence of regulatory T cells and the production of anti-inflammatory cytokines. Remarkably, no systemic toxicity was observed. This study is the first scientific proof of antitumor potential of A. vulgaris extract in vivo.

Alchemilla vulgaris agg., apoptoza, autofagija, imunski odgovor, invazija, melanom, migracija, mikrosredina tumora, tumor dojke

Alchemilla vulgaris agg., apoptosis, autophagy, immune response, invasion, melanoma, migration, tumor microenvironment, breast tumor

577.27:618.19‑006(043.3)

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