The objective of our studies were seeds of two lupin species Lupinus luteus L. and Lupinus angustifolius L. cvs. Lord and Graf respectively. Lupin seeds were germinated at 15 and 24°C and during two, three and four days. In the lupin sprouts antinutritional factors: alkaloids and raffinose family oligosaccharides (RFOs) and five nitrogen fractions: non protein (Nnp), albumin (A), globulin (G), glutelin and prolamin (Gt+P) and nitrogen residue fraction (Nr) were determined. The level of these compounds was compared with the proper ones of initial material (not germinated seeds). These studies showed that the germination process clearly affects the decrease of antinutritional factors: RFOs and alkaloids. The decrease level of these compounds depended on such factors like, lupin species and used germination conditions. It was found on the base of nitrogen analysis of particular protein fractions that the germination process of lupin seeds causes deep quantitative and qualitative changes in fractional composition of lupin proteins. It especially concerns the decrease of globulin and residual fraction content and distinct increase of Nnp fraction. The changes in other fractions were not so unequivocal in comparison with the mentioned above and depended on lupin species, temperature and time of germination. Qualitative changes of A, G and Gt+P fractions caused by germination were confirmed by gel electrophoresis (SDS-PAGE). The amino acid analysis of seeds and sprouts of Nnp fractions showed an increased content of Asp, Ser, Ala, Pro non essential amino acids (NEAA), and Val, Met, iLeu, Leu, Thr essential amino acids (EAA). Simultaneously a decrease of Glu, Arg (NEAA), Phe, Lis, Cys (EAA) contents was observed. Generally the germination process causes the decrease of total NEAA and an increase of total EAA in Nnp fractions of both lupin species.

Wound fluids (WF) are believed to play a role in the local recurrences by inducing an inflammatory process in scar tissue area. Given that most local relapse in primary breast cancer patients occur within the scar tissue area, researchers have investigated whether localized radiotherapy, such as intraoperative radiotherapy (IORT), could be more effective than postoperative RT in inhibiting local tumor recurrence. The epithelial-mesenchymal transition (EMT) program plays a critical role in promoting metastasis in epithelium-derived carcinoma. Given this background the main aim of the present study was to determine the mechanisms by which IORT decreases the tumorigenic potential of WF. We assumed that postoperative fluids from patients would activate the radiation-induced bystander effect (RIBE) in treated cells, thus altering the tumor microenvironment. To confirm this hypothesis, WF collected from patients after breast conserving surgery (BCS) alone, after BCS followed by IORT treatment or WF from BCS patients together with RIBE medium were incubated with MCF7 and MDA-MB-468 cells. Changes in the CSC phenotype, in EMT program and potential to migrate were performed to determine the possible role of WF on the migration of breast cancer cells. Our findings show that wound fluids stimulate the CSC phenotype and EMT program in breast cancer cell lines. This effect was partially abrogated when the cells were incubated in wound fluids collected from patients after breast-conserving surgery followed by IORT. Additionally, we confirmed the role of radiation-induced bystander effect in altering the properties of the WF to induce the CSC phenotype and EMT program.

Breast cancer is the most common cause of skin metastases in women. The probability of their occurrence ranges from about 5% in the entire population to as much as 30% in the late stages of the disease. Although rarely life-threatening, they have a major impact on the quality of life of patients with this diagnosis, being the cause of pain, effusion, ulceration, infection, and psychological discomfort. Available methods of treatment, both local and systemic, often fail to provide adequate control of the disease. A particular challenge seems to be the treatment of those patients with cutaneous metastases who, due to the extent of their metastases, are not eligible for resection, in whom the possibility of radiation therapy has already been used, and in whom systemic therapy is ineffective or contraindicated. A new method providing the opportunity for effective treatment is electrochemotherapy (ECT). ECT combines electropulsation of tumor cells (by local application of electric pulses) and administration of antineoplastic drugs such as cisplatin or bleomycin (either intravenous or intratumoral). Several clinical studies have demonstrated that ECT provides safe, efficient, and non-invasive locoregional treatment for chest wall breast cancer recurrence.

The wound healing process after surgery alters the area surrounding the original tumor and around the scar, and the modified microenvironment is more favorable for tumor recurrence. Intraoperative radiotherapy (IORT) is one of the more novel strategies in breast cancer (BC) treatment. Irradiation during surgery has effects on the tumor microenvironment, abrogating the proliferative cascade induced by surgical wound healing. The aim of the present study was to determine the effect of surgical wound fluids from IOERT treatment (RT-WF) compared with wound fluids from conservative-breast surgery only (WF) on the cancer stem cell phenotype in a panel of BC cell lines. Post-operative wound fluids were derived from patients with BC who underwent a tumor resection (quadrantectomy) plus intraoperative electron radiotherapy using a single dose of ≤10 Gy on the tumor bed and surrounding tissues, or from those who underwent a tumor resection without IOERT. Cell lines were incubated with 10% wound fluids, and after 4 days, the cluster of differentiation (CD)44+/CD24−/low phenotype and aldehyde dehydrogenase 1 (ALDH1) activity were determined by flow cytometry. The two types of fluid each affected the CD44+/CD24−/low phenotype. The results varied markedly between each cell line, even for the same histological subtypes. RT-WF decreased the CD44+/CD24−/low populations in the basal-like BT-549 and MDA-MB-468 cell lines, whereas in the luminal type MCF7 cell line, the two fluids inhibited these populations. The HER-OE subtypes harbored a minimal CD44+/CD24−/low population, but the growth of SK-BR-3 was stimulated by the two post-operative fluids. WF exhibited a stronger effect on ALDH1 activity compared with RT-WF. The stimulatory effect was dependent on the histological subtype of the cell line and the strongest dependence was observed in luminal subtypes characterized by low dehydrogenase activity in the control group. The present results enable a better understanding of the mechanism of recurrence and metastases following BC surgery. With respect to histological phenotype, its effect on tumor progression, either local or systemic, strongly suggests the requirement for further research and clinical validation.

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Overall, tumor micromilieu and tumor growth could not be changed by glycolysis modifiers in the three tumor cell lines in vivo. Further studies are necessary to explore the impact of metabolic targets on radiation response.

Breast cancer is the most common malignant disease occurring in women. Conservative breast cancer surgery followed by radiation therapy is currently the standard treatment for this type of cancer. The majority of metastases occur within the scar, which initiated a series of studies. As a result, clinical trials aimed to assess whether localized radiotherapy, as intraoperative radiotherapy (IORT), may more effective in inhibiting the formation of local recurrence compared with the standard postoperative whole breast radiotherapy. The present study determined the role of postoperative wound fluids (WFs) from patients diagnosed with breast cancer subsequent to breast conserving surgery or breast conserving surgery followed by IORT on the expression of three microRNAs (miRNAs), consisting of miR-21, miR-155 and miR-221, in distinct breast cancer cell lines that represent the general subtypes of breast cancer. It was determined that the miRNAs responsible for breast cancer progression, induction of tumorigenesis and enrichment of the cancer stem cell phenotype, which is responsible for resistance to tumor therapy, were highly upregulated in the human epidermal growth factor receptor 2-positive breast cancer SK-BR-3 cell line following stimulation with WFs. It is worth emphasizing, that those changes were more significant in WFs collected from patients after surgery alone. The BT-549 cell line showed altered expression only of miR-155 following incubation with WFs. Notably, this change was not associated with IORT. Additionally, it was indicated that both WFs and RT-WF strongly downregulated the expression of miR-21, miR-155 and miR-221 in basal/epithelial and luminal subtypes of breast cancer. It was concluded that the present study contributes to an increased understanding of the role of surgical WFs and IORT treatment in the regulation of miRNA expression. This may enable the development of the current knowledge of breast cancer biology subsequent to IORT treatment and substantially to improve the therapy in the future.