Searching for combination therapy by clustering methods: Stimulation of PKC in Golgi apparatus combined with hypericin induced PDT

Selection of Unique Molecules for Cancer Treatment by Distance-Based Method: Hypericin Effect on Respiratory Chain

“…Cell lysates were prepared in RIPA buffer (150 mM NaCl, 1% Triton X-100, 0.5% sodium deoxycholate, 0.1% sodium dodecyl sulfate, 20 mM Tris buffer saline at pH 8.0) with inhibitor cocktail (2x 1:100, HaltTM Protease & Phosphatase Inhibitor Cocktail, ThermoFisher Scientific, Waltham, MA, USA) from cells incubated for 1 h with 0.5 µM Gö 6976, 0.5 µM PMA, 10 µM rottlerin, and 0.5 µM hypericin in 10% FBS or 2% UG. Electrophoresis was performed according to the protocol described previously [6]. Proteins were transferred to a nitrocellulose membrane.…”

“…The literature and references from our previous publications [6,[8][9][10][11][12][13][14][15][16][17][18][19][20][21][22][23][24] provide us with quantitative information that suggests a possible classification. We first categorize and standardize the obtained knowledge into two main areas to investigate the topic using our chosen methods.…”

“…For this reason, a greater effort is devoted to multiscale analysis of the problem [32,33]. As shown in our recent research [6], the induction of PKC activation and phosphorylation depends on the lipid content, and lipid metabolism could be controlled with the hydrophobic molecules used. According to the heterogeneous distribution of lipids in the tumor, the cell response and signaling pathways triggered by PKC could also be heterogeneous.…”

“…The problem of analysis, as well as comparing the stabilizing or destabilizing effect of different types of stimuli leading to either survival or cell death, can be applied to the specific case of protein kinase C (PKC) regulation. In this context, we have already identified four regulators of PKC: Gö 6976, PMA, rottlerin, and hypericin [6]. The evaluation can be based on a reduction to three levels of molecular effects, e.g., to discrete values of 0, 1, and 2.…”

“…There is some continuity of development in our research steps, which we briefly summarize here. We have shown that distance-based approaches, specifically cluster analysis of fluorescence lifetime probability vectors, can be used to reliably assess the extent of oxidative stress to glioma cells based on differences in sensor fluorescence lifetimes between five different treatments [6]. A recent study investigated statistical fluorescence lifetime abnormalities in oxygenation and oxidative stress in cells and tissues using different spacing models, which then served as the basis for hierarchical clustering approaches [7].…”