Double face of cytochrome c in cancers by Raman imaging

Abstract

Cytochrome c (Cyt c) is a key protein that is needed to maintain life (respiration) and cell death (apoptosis). The dual-function of Cyt c comes from its capability to act as mitochondrial redox carrier that transfers electrons between the membrane-embedded complexes III and IV and to serve as a cytoplasmic apoptosis-triggering agent, activating the caspase cascade. However, the precise roles of Cyt c in mitochondria, cytoplasm and extracellular matrix under normal and pathological conditions are not completely understood. To date, no pathway of Cyt c release that results in caspase activation has been compellingly demonstrated in any invertebrate. The significance of mitochondrial dysfunctionality has not been studied in ductal carcinoma to the best of our knowledge. We used Raman spectroscopy and imaging to monitor changes in the redox state of the mitochondrial cytochromes in ex vivo surgically resected specimens of human breast tissues, and in vitro human breast cells of normal cells (MCF 10A), slightly malignant cells (MCF7) and highly aggressive cells (MDA-MB-231). We showed that Raman imaging provides insight into the biology of human breast ductal cancer. Here we show that proper concentration of monounsaturated fatty acids, saturated fatty acids, cardiolipin and Cyt c is critical in the correct breast ductal functioning and constitutes an important parameter to assess breast epithelial cells integrity and homeostasis. We look inside human breast ducts by Raman imaging answering fundamental questions about location and distribution of various biochemical components inside the lumen, epithelial cells of the duct and the extracellular matrix around the cancer duct during cancer development in situ. Our results show that human breast cancers demonstrate a redox imbalance compared to normal tissue. The reduced cytochrome c is upregulated in all stages of cancers development. The results of the paper shed light on a largely non-investigated issues regarding cytochromes and mitochondrial function in electron transfer chain. We found in histopathologically controlled breast cancer duct that Cyt c, cardiolipin, and palmitic acid are the main components inside the lumen of cancerous duct in situ. The presented results show direct evidence that Cyt c is released to the lumen from the epithelial cells in cancerous duct. In contrast the lumen in normal duct is empty and free of Cyt c. Our results demonstrate how Cyt c is likely to function in cancer development. We anticipate our results to be a starting point for more sophisticated in vitro and in vivo animal models. For example, the correlation between concentration of Cyt c and cancer grade could be tested in various types of cancer. Furthermore, Cyt c is a target of anti-cancer drug development and a well-defined and quantitative Raman based assay for oxidative phosphorylation and apoptosis will be relevant for such developments.

The effect of COVID-19 mRNA vaccine on human lung carcinoma cells in vitro by means of Raman spectroscopy and imaging

“…It is worth of noticing that a similar effect of down regulation of cytochrome c concentration was reported for brain cancer cells vs. cancer aggressiveness [9,15]. We showed that the Raman signal of the band at 1584 cm -1 related to the concentration of cytochrome c in mitochondria of a single cell in vitro decreases with brain tumor aggressiveness [10,15].…”

“…The peak at 1584 cm -1 in Figure 2 represents the “redox state Raman marker” of cytochrome c. Recently we demonstrated that this Raman vibration can serve as a sensitive indicator of oxidized and reduced forms of cytochrome c [9,10,12]. It indicates that the Raman peak at 1584 cm -1 can be used as a marker to explore apoptosis and oxidative phosphorylation in mitochondria [16,18].…”

Hyperglycemia and Cancer. Human lung carcinoma by means of Raman spectroscopy and imaging

“…The Raman intensity of the oxidized form is much smaller than that of the reduced form. [34] [26] [44] [25] Inside a normal mitochondrium cytochrome c exists in the oxidized form. Dysfunction of mitochondrium associated with several malignancies, including cancer or hyperglycemia blocks the transfer of electrons between complexes III and IV of the respiratory chain, resulting in decreasing the efficiency of the oxidative phosphorylation (respiration) process and lower ATP synthesis.…”

“…We will concentrate on de novo lipogenesis, which is being increasingly recognized as a hallmark of cancer. [32] [33] [34]…”

“…We will concentrate on de novo lipogenesis, which is being increasingly recognized as a hallmark of cancer. [32] [33] [34] The aim of this paper is to demonstrate the possibility of monitoring glucose changes occurring in cancer cells by Raman spectroscopy and Raman imaging. The aim of this work is to understand the role of sugars in metabolism of human lung cancer cell lines CCL-185.…”