Apoptosis Induced by Culturing MH134 Cells in the Presence of Porcine Skin Gelatin In Vitro

Abstract

Porcine skin (PS) gelatin showed antitumor effect in vitro on MH134 murine hepatoma cells. We analyzed the effect of PS gelatin on MH134 cells compared to the effect of Bovine bone (BB) gelatin, 5-Fluorouracil (5-FU) and 7-ethyl-10-hydroxycamptothecin (SN-38). We previously suggested that PS gelatin induces apoptosis in MH134 cells using flow cytometric analysis. We performed agarose gel electrophoresis and electron microscopic studies to ascertain apoptosis. Agarose gel electrophoresis showed the typical DNA ladder pattern and electron microscopic findings revealed characteristic features in the case of apoptosis on PS gelatin. SN-38 also showed DNA ladder pattern and ultrastructural changes in apoptosis. 5-FU didn't show DNA ladder pattern but electron microscope revealed changes in necrosis. On the other hand, BB gelatin didn't induce apoptosis or necrosis.

In vitro impact of a whey protein isolate (WPI) and collagen hydrolysates (CHs) on B16F10 melanoma cells proliferation

“…Ito et al [8] demonstrated in vitro that porcine skin (PS) gelatine induced apoptosis in MH134 cells evaluated through flow cytometry, agarose gel electrophoresis and electron microscopy. On the other hand, bovine bone (BB) gelatine did not induce apoptosis or necrosis.…”

“…On the other hand, bovine bone (BB) gelatine did not induce apoptosis or necrosis. Probably, these differences resulted from different primary sequence of amino acids in these collagen molecules [8].…”

“…Collagen has already shown bioactivities regarding inhibition of gastric ulcer lesions [28][29][30], tumorigenesis [4][5][6]8], suggesting its possible function in extracellular matrix influencing the intra and extra cellular signals and differentiation, genetic changes, stimulation of specific enzymatic cascades, apoptosis as well as other molecular and cellular mechanisms [23]. Inhibitory effect of fibrillar collagen on human melanoma and fibrosarcoma cells proliferation involving activation of the tyrosine kinase discoidin domain receptor 2, which is independent of effects on cell spreading, promoted growth arrest in the G0/G1 phase of the cell cycle [6].…”

“…Annexin V is a phospholipid-binding protein with high affinity for phosphatidylserine, what facilitates the identification of apoptotic cells with exposed phosphatidylserine. Necrotic cells are promptly stained with PI as well as with annexin V whereas apoptotic cells only stains with annexin V [8].…”

“…Ito et al [8] demonstrated in vitro, that porcine skin gelatine presented anti-tumoral effect on MH134 murine hepatoma cells, inducing programmed cell death (apoptosis).…”