Breast cancer is the most common cancer type affecting women worldwide [1]. It has been the leading cause of cancer in women in Europe for many years, but survival rates are high [2]. Despite these successes, new technologies are needed for the treatment of all stages of breast cancer, not only to improve outcomes, but also especially to provide more options for all patients who are not suitable or who do not respond to standard treatment.

A new therapeutic modality that may give an advantage over surgery in patients who are not suitable for conventional treatments is photodynamic therapy (PDT) [3]. Photodynamic therapy is based on the reception of a photosensitive photosensitizer (PS) by the tumor and its interaction with light [4]. PS causes cellular damage by producing singular oxygen (singlet oxygen) after interaction with light of the appropriate wavelength and in the presence of oxygen [5].

Aza-BODIPYs, boron complexes of aza-dipyrromethenes, have absorption and emission above 650 nm in the visible region. The effects of phthalonitrile presence on the photophysical and photochemical properties of the aza-BODIPY structure depend largely on the types of substituents [6]. In our previous study, aza-BODIPY dye carrying phthalnitrile groups in the para positions of distal phenyl groups (aza-BODIPY), was prepared and the photophysical, photochemical and photobiological properties of this aza-BODIPY derivative were studied. Furthermore, the cytotoxicity of this aza-BODIPY derivatiev against breast cancer cell line (MCF-7 cells) was investigated in our previous study [7,97]. On the cancer cell line MCF-7, testing for the aza-BODIPY compound's under dark and light conditions were conducted [7,97].

It has been demonstrated that using PDT and particular PSs successfully kill cancer cells [8]. The underlying mechanism of cell death induction must be identified in order to evaluate the effectiveness of PDT utilizing particular PSs. Breast cancer is a complex disease that can present with different levels of aggressiveness, making it difficult to treat. There is still no clear explanation of how PDT destroys tumor cells in breast cancer. However, it remains a matter of curiosity where PDT, PS, are localized within the cell and how they affect cytotoxicity. Despite the fact that it is believed that MCF-7 cells utilize various molecular cancer biology pathways, there are gaps in the literature on this topic. Therefore, in this study, we evaluated the carcinoma and cell death mode pathways modulated by aza-BODIPY PDT of human breast cancer.

Various signaling pathways are involved in normal breast development in the organism. These include mainly the Wnt/β-catenin pathway, the estrogen receptor signaling pathway, and the HER2 signaling pathway [9].

In this study, we investigated the effect of aza-BODIPY-PDT on the expression of 88 carcinoma-associated genes in the breast cancer cell line MCF-7 using the GeneQuery™ Human Basal Cell Carcinoma qPCR Array Kit. We have provided a brief account of the significance of the genes analyzed in this study below

The first gene category is the APC gene, which by constitutively boosting the Wingless/Wnt signal transduction pathway, can cause β-catenin to accumulate in the cytoplasm and abnormal cellular proliferation. All of these things are thought to contribute to APC's inhibition of the Wingless/Wnt signaling pathway. The Wingless/Wnt signaling pathway is emerging as a new therapeutic target for the treatment of triple-negative breast cancer, which refers to ER, PR, and HER 2-negative breast cancer [10].

And also APC and APC2 are both expressed in human breast epithelium, and both may be implicated in the control of β-catenin/Wnt signaling [11]. Loss of heterozygosity somatic mutation, promoter hypermethylation in breast cancers, and other mechanisms have been shown to reduce APC [12].

The other gene group is CD151; family of integrin-associated cell surface molecules, through research on tetraspanins, it has also been possible to determine the functional connection between integrins and Wnt signaling in breast cancer [10]. Members of the tetraspanin family, such as CD151, CD9, CD82, and TSPAN12, have all been discovered to have a part in regulating carcinogenesis in relation to Wnt signaling [13].

In addition to these gene families, the screening approach revealed Casein kinase 1 gamma 2 (CSNK1G2), a serine-/threonine protein kinase, to be the most sensitive target to TAM. Breast cancer cells that express the estrogen receptor (ER+) were highly cytotoxicly impacted, whereas ER- cells were just marginally toxicated. Additionally, CSNK1G2 knockdown greatly decreased tumor spheroid formation and the expression of breast stem cell marker genes including CD44/CD2 in ER+ breast cancer cells [14].

Another crucial gene class is IF4E, which is overexpressed in a variety of human cancers. Experimental model [15] have revealed that the gene [16] acts as an oncogene. Breast cancer patients with poor prognoses have been found to have a higher recurrence rate in both node-negative and node-positive patients, and increased levels of the proteins cyclin D1, c-Myc, and VEGF have been associated to overexpression of eIF4E in this disease [17,18].

A different class of genes thought to be associated with cancer is FZD5; The Frizzled family member FZD5, which is selectively expressed in TNBC and is connected to a poor prognosis for the condition, has been identified [19].

Glycogen Synthase Kinase-3 (GSK-3) is a serine/threonine protein kinase that has been identified as a crucial enzyme involved in glycogen metabolism, but is now recognized as a regulator of multiple cellular activities. It is one of an important class of genes suspected to be related with breast cancer. By phosphorylating a variety of metabolic, signaling, and structural proteins, GSK-3 helps to control its activity [20]. Numerous prognostic indicators are associated with the overexpression of GSK-3 in human breast cancer, and patients with breast cancer who have GSK-3 expression levels in the highest quartile (246 of 1686 cases) are at increased risk of distant recurrence 5 and 10 years after tumor excision, with increases of 7 and 1.7 times, respectively [21].

A different class of gene is LRP2 (megalin), the endocytic membrane receptor, is known to be necessary for the endocytosis of numerous ligands in specialized epithelium, including the thyroid gland, mammary glandular epithelium, and the proximal tubules of the kidney. Furthermore, it is yet unknown how LRP2 is regulated and what role it plays in malignancies coming from these tissues [22].

A new target for the treatment of cancer is the Rab23 gene, which is thought to be a molecule that adversely affects the hedgehog (Hh) signaling pathway. According to a study, Rab23 is produced by breast cancer cells and, in addition to causing cell death, ectopic Rab23 expression stops breast cancer cells from multiplying and developing. The inhibition of Gli1 and Gli2 mRNA expression by Rab23 is thought to be the cause of these findings. According to these findings, Rab23 may be a viable target for the therapy of breast cancer [23].

Moreover Tcf7l1, a crucial Wnt/β catenin signaling pathway effector molecule, is highly expressed in a variety of malignancies and encourages tumor growth [24]. Breast cancer [25], colorectal cancer [26], squamous cell carcinoma tumors on the skin [27], and colorectal cancer tumors [28] have all been documented to have high expression levels of Tcf7l1, which promotes tumor growth. Although the majority of these findings concentrated on Tcf7l1′s effects on tumor proliferation, Slyper et al. demonstrated that Tcf7l1 improves breast cancer cells' capacity to form spheres and aids in their capacity to self-renew [25].

WNT ligands are an additional important protein subgroup. Wnt signaling is a highly conserved signaling mechanism that regulates the development of cancer in addition to embryonic and organ development. Wnt signaling is mainly involved in the growth and metastasis of breast cancer, according to assessments of gene expression profiles and genome-wide sequencing. Three well-known Wnt signaling pathways—Wnt/ β-catenin, Wnt-planar cell polarity (PCP), and Wnt-Ca2+ signaling—have overlapping elements but serve distinct functions in the development of breast cancer. The modulation of the immunological microenvironment in breast cancer, trunk maintenance, treatment resistance, phenotypic sculpting, etc. are all demonstrated to be impacted by Wnt signaling in recent studies [29].

A variety of β-catenin-independent, non-canonical WNT signaling cascades can be started by WNT ligands like WNT5A/B or WNT11 binding to FZDs and alternative co-receptors; the most studied of these are the WNT/Calcium (Ca2+) and WNT/planar cell polarity (PCP) signals. The PKC, NF-kB, and CREB are activated as a result of an increase in intracellular calcium levels that characterizes the WNT/Ca2+ pathway [30].

Numerous tumor cells, such as those from cervical, breast, prostate, and colon cancers, have been shown to proliferate and migrate more when WNT11 is overexpressed [31]. These findings reveal that WNT11 signaling is essential for cancer cell proliferation, invasion, and metastasis [32].

According to one study, Wnt5b is a crucial regulator that controls the Basal-like breast cancer (BLBC) phenotype by triggering both canonical and non-canonical WNT signaling. Wnt5b is a possible treatment target for BLBC in addition to being a diagnostic biomarker [33].

WNT16 was identified as the gene increased in carcinoma-associated fibroblasts (CAFs) following chemotherapy based on the findings of a microarray investigation [34]. Because of these factors, it is possible that chemotherapy-induced WNT16 expression in CAFs is linked to breast cancer treatment resistance [35].

E-cad protein is necessary for the growth and upkeep of polarized and differentiated epithelium via intercellular adhesion complexes. In more severe cases of some epithelial carcinomas, dysregulation of this molecule, which is thought to be an invasion suppressor, is frequently seen. It has been discovered that normal E-cad function prevents the spread of breast cancer. Somatic E-cad inactivation has been related to an aggressive pattern of breast cancer, involving lymphovascular invasion and metastases in axillary lymph nodes, according to publications [36], [37], [38].

Besides these FAT2 gene group is also important for cancer patients. Patients with FAT2, which is highly expressed in lung squamous cell carcinoma and breast cancer, have a bad prognosis. Overall survival in patients with FAT2 mutations has been shown to be significantly worse in esophageal squamous cell adenocarcinoma and carcinoma [39]. FAT2 mutation in somatic cells is suggested to encourage metastasis in colorectal cancer [40].

One of the genes associated with cancer is FZD6. It has been discovered that Frizzled receptors mediate Wnt ligand signaling, which is typically produced in cancer and is crucial for controlling tissue development and differentiation. According to one study, the triple negative breast cancer (TNBC) subtype is more likely to have the Wnt receptor frizzled 6 (FZD6) gene amplified than other subtypes of breast cancer [41].

Hhat belongs to a distinct gene class that has been linked to breast cancer. In a study, it was discovered that HAT is necessary for the development of ER positive, HER2 positive, and tamoxifen-resistant breast cancer cells. According to studies, ER positive cells proliferate more when Hhat is expressed more, while ER positive cells proliferate less when Hhat is depleted [42]. A separate research revealed that inhibiting Hhat with the selective small molecule inhibitor of Hhat, RU-SKI 43, also inhibited the development of ER positive cells [43].

The other gene is MC1R, The G-protein coupled receptor MC1R plays a role in the etiology of melanoma by causing UV light-induced melanin production and DNA repair in melanocytes. According to studies, breast cancer tissue has significantly greater levels of MC1R expression than healthy breast tissue, and both in vitro and in vivo MC1R downregulation significantly reduced cell proliferation [44].

Additionally, the Wnt signaling pathway includes transcription factor 7-like 2 (TCF7L2), also known as T cell factor 4 (TCF4), which is situated on 10q25.2 [45]. Tumor onset and growth have been discovered to be strongly correlated with the activation of the Wnt signaling system. The association between TCF7L2 polymorphisms and breast cancer susceptibility is quite intriguing given the available evidence. TCF7L2 genetic mutations have been linked to an increased risk of breast cancer, according to analyses [46].

One of the important protein is AXIN1. AXIN1 is a multi-domain scaffold protein with tumor-suppressive properties; it primarily acts as a rate-limiting component in the complex that breaks down β-catenin [47]. Other than its well-known role as a negative regulator of Wnt/ß-catenin signaling [48].

C/EBPa is another gene that has been linked to cancer. The subfamily's founding member is the critical region leucine zipper (bZIP) transcription factor CCAAT/enhancer-binding protein alpha (C/EBPa) [49].It regulates the expression of numerous genes that are unique to distinct lineages as well as the cell cycle [50]. C/EBPA gene changes are present in about 10 % of instances of acute myeloid leukemia (AML), demonstrating the tumor suppressive role of C/EBPa in the development of cancer [51]. Additionally, extensive sequencing studies have shown that C/EBPa is one of 125 genes that could be impacted by intragenic changes to cause cancer [52]. Although deregulated C/EBPa expression has been observed in a number of solid tumors, such as liver, breast, and lung cancer, its relationship to carcinogenesis is yet unclear. An investigation of C/EBPa expression in healthy breast tissue and breast carcinomas [53] revealed that the majority of breast cancer samples had decreased levels of C/EBPa.

The CTNNB1 gene, which is found on chromosome 3p22.1, also encodes the -catenin protein in addition to these. It plays a significant role in the cell-to-cell contacts of the Wnt signaling pathway and is essential for the transcriptional regulation of that system by connecting cadherins to the actin cytoskeleton. The importance of CTNNB1 activating mutations and β-catenin dysregulation in the emergence of cancer is supported by a number of lines of evidence [54,55]. The majority of research has concentrated on the issue of the failure of the β-catenin/Wnt pathway in breast cancer [56,57].

FBXW11 is a crucial component of the genes that we looked at. According to results from earlier studies, FBXW11 stimulates the signaling pathways for nuclear factor-kB (NF-kB) and β-catenin/T-cell factor at the same time to promote the proliferation of lymphocytic leukemia cells [58].

Another gene is FZD7. According to studies, the Fzd7/beta-catenin pathway, which promotes cell proliferation, is also active in triple-negative breast cancer (TNBC) [59].

The other gene group is Hhip, endothelial cells exhibit large quantities of Hhip, an inhibitor of the hh pathway, although epithelial and fibroblastic cells also express hhip at significantly lower levels, during angiogenesis, and in tumor tissues [60].

Another gene class known as Net1 is thought to have a link to cancer. One of the RhoA subfamily GEFs known to be overexpressed in a variety of human malignancies, including breast cancer, is the neuroepithelial transforming gene 1 (Net1) [61]. An earlier investigation found that Net1 is necessary for the human breast cancer cells' motility and in vitro invasiveness [62].

The SHFM1 gene, which causes split hand/foot malformation (ectrodactyly) type 1, may be crucial for preserving genomic integrity. Its biological action is said to make it a potential participant in hereditary cancer susceptibility. The SHFM1 protein binds to the longest conserved region of the BRCA2 protein in mammalian cells, which is essential for BRCA2′s stability and functionality as well as for its function as a mediator of homologous recombination. Due to all of these circumstances, variations in the SHFM1 gene may reduce BRCA2 function and be connected to the familial development of breast/ovarian cancer [63].

Important TGF-ligands (TGFB1, TGFB2, and TGFB3) work with their receptors to activate signal transduction. Transmembrane serine/threonine protein kinase receptor TGFBR2, to which the ligands mostly bind; TGFBR3 may sporadically facilitate this interaction (ana makale). This complex binds to and activates TGFBR1 following the phosphorylation of SMAD2 and SMAD3. It is believed that genetic diversity in the TGF-beta signaling pathway's constituent parts can affect how proteins function, whether target genes are expressed more or less, and how breast cancer develops and spreads [64].

Additionally, Hh ligands (Sonic-SHH, Indian-IHH, and Desert-DHH) are proteins that attach to the Patched (PTCH) cell surface transmembrane receptor. Smoothened (SMO), a transmembrane receptor-like protein, is suppressed by PTCH, but when it attaches to ligands (SHH, IHH, or DHH), it releases SMO, which in this case causes processing to stop GLI (glioma-associated oncogene homolog)–zinc-finger transcription factors. Three mammalian GLI proteins have so far been identified. While GLI3 plays the opposing function, GLI1 and GLI2 frequently act as transcriptional activators. Small molecule smoothened (SMO) inhibitors like vismodegib have been effective in treating basal cell carcinomas that have Hh pathway mutations like PTCH1. A number of different malignancies, including lung cancer, colorectal cancer, prostate cancer, breast cancer, and malignant melanoma, have been linked to additional pathways of aberrant activation, including as overexpression of Hh ligand, and autocrine and paracrine signaling [65].

Besides, Type I IFNs have a role in the regulation of the immune system's response to cancer and have strong anticancer properties. Despite having favorable effects in the treatment of cancer, type I IFNs' precise mechanisms of immune surveillance against tumors are still unknown. It has been demonstrated that type I IFNs are essential for enhancing host immune responses against cancer through the activation of a range of immune cells, including T cells, natural killer (NK) cells, DCs, and macrophages. When the results of recent studies were reviewed, it became clear that type I IFNs also have an impact on neutrophil activation and support the anticancer capabilities of these cells [66].

On the other hand, PARP-1, has drawn a lot of interest as a possible target for pharmacological inhibitors in the treatment of breast cancer. A study discovered that the luminal subtype of breast cancer had greater levels of the estrogen-protected gene set and PARP-1 expression, and that high PARP-1 expression was linked to a poor prognosis in ER+ patients [67].

In that regard, SHH, PTCH1, and GLI1 mutations were extremely uncommon in BC [68], preventing mutational activation of the Hh pathway in BC. It has been discovered that ligand-dependent stimulation of Hh signaling via overexpression of SHH or IHH is linked to a number of malignancies [69].

In this situation, breast cancer will manifest in up to 85 % of women with germline TP53 gene mutations by the time they reach the age of 60. With a mean diagnosis age of 34, the majority of these breast cancers present at an early stage. It has been determined that a germline TP53 gene mutation affects between 5 and 8 percent of breast cancer patients under the age of 30. Breast cancers in women with TP53 mutations are reportedly more likely to be Her2 positive and/or hormone receptor positive [70].

In this line, The TGF-β superfamily of growth factors includes multifunctional growth factors known as bone morphogenetic proteins (BMPs). Studies have shown that BMP-2 enhances the motility and invasiveness of breast cancer cells both in vitro and in the mouse xenograft model, as well as the epithelial-mesenchymal transition (EMT) [71].

New data suggests that DVL is also essential for breast carcinogenesis. A recent study revealed that TNBC samples have higher DVL-1 protein expression levels than non-cancer samples [72].

PRKAR1A is the main regulatory subunit of PKA and the main its catalytic subunit is PRKACA. Unexpectedly, recent studies also revealed that the expression of the PRKAR1A protein was significantly dysregulated in a variety of primary carcinomas and distant metastases, including, breast cancer, pediatric pituitary adenomas [73].

Additionally, SMO, another crucial gene linked to cancer. Smoothened (SMO) is a component of the Hedgehog (HH) signaling system, which regulates cancer stem cells, cell migration, invasion, and proliferation. The HH signaling pathway includes both canonical and non-canonical pathways. Cancers of the breast, liver, pancreas, and colon are only a few of the tumors for which the role of SMO has been studied. SMO expression has been connected to tumor invasion, metastasis, recurrence, and expansion [74].

Likewise thioredoxin-1 (Trx1, TXN), is abundantly expressed in a variety of cancers and affects cell growth and death through various signaling pathways [75]. The significant tumor suppressor effects of small molecule thioredoxin system inhibitors [76] suggest that TXN may be a promising therapeutic target in the treatment of cancer.

In patients with solid tumors, expression of TXN has been linked to rapid tumor development and decreased survival. TXN is regarded as a viable target for the development of anti-cancer drugs since it is crucial for maintaining the altered phenotype and treatment resistance of various human malignancies. TXNR is also known to be involved in chemoresistance and is increased in many cancer cells [77].

On the side, a growth factor that is a member of the bone morphogenetic protein (BMP) family, which makes up the majority of the transforming growth factor (TGF-Beta) superfamily, is known as bone morphogenetic protein 4 (BMP4). BMPs act as serine/threonine receptor-binding extracellular ligands that communicate with cells via intracellular SMAD mediators as well as other routes, like the MAP kinase pathway, and serine/threonine receptors on the cell membrane. BMPs were initially identified for their ability to promote the formation of bone, but further research has revealed that they are also powerful developmental regulators. BMP4, for instance, is essential for the development of many organs and tissues, such as the mammary gland, hematopoiesis, gastrulation, and mesoderm formation [78]. BMPs are increasingly being investigated as potential participants in cancer because of their multifunctionality. Depending on the tissue of origin, BMP4 expression has been shown to vary in cancer and may even increase or decrease [79]. Strong BMP4 expression has been seen in breast cancer cell lines and tissues, and immunohistochemistry research has shown that BMP4 protein is present in 25–50 % of original tumors [80].

GAS1 is the other gene family. The growth arrest-specific (Gas1) gene, which encodes a glycosylphosphatidylinositol-associated protein, was found as a result of a search for genes that are selectively expressed in growth-arrested cells. Just before the cell cycle's S phase, it was discovered that overexpression of this gene causes cell arrest. It has been suggested that the human GAS1 gene, which is located in the often deleted chromosomal regions 9q21.3-q22, may play a role in the prevention of tumor cell proliferation [81].

Furtermore, metastatic renal cell carcinoma has been linked to significant overexpression of a number of important IFN signaling pathway effectors, including IFNGR1, IFNGR2, STAT1 and STAT2 [82]. The STAT2 homodimer can promote IL-6 gene expression either directly or as part of the ISGF3 complex. Due to decreased levels of inflammatory cytokines, STAT2 absence was found to inhibit colorectal carcinogenesis in a STAT2 knockout animal study [83].

Beside these, the SCFTrCP E3 ubiquitin ligase's component for substrate identification is TrCP, a protein belonging to the family of F-box proteins. It has two paralogs, BTRC (also called TrCP1) and FBXW11 (also called TrCP2 or Hos). BTRC and FBXW11 are physically and functionally similar despite being encoded by separate genes. The release of NF-kB dimers into the nucleus is facilitated by recognition by TrCP, which causes IκBα ubiquitination and then induces proteasomal destruction of the substrate [84,85].

Furthermore, the high-mobility transcription factor T-cell factor (TCF)/LEF1 lymphoid enhancer-binding factor 1 (LEF1) is primarily involved in the Wnt/β-catenin signaling pathway. LEF1 is overexpressed in lung adenocarcinomas and oral squamous cell carcinomas, according to several analyses, and its aberrant expression is directly linked to tumor progression and a poor prognosis [86]. Also, Sufu is a crucial critical suppressor of the Hh pathway, when inappropriately active promotes carcinogenesis [87].

In this regard, damaged apoptosis is one of the characteristics of cancer. In cancer, the calpain family of cysteine proteases has been shown to interact with caspase-3 and caspase-8, two important regulators of the apoptotic response [88]. Increased caspase-3 expression has been linked to a significantly better prognosis in patients with hepatocellular carcinomas and non-small cell lung cancer, according to earlier research [89].

As well as, the epidermal growth factor (EGF) receptor (EGFR) pathway serves as a good illustration of a signal transduction pathway that is linked to the onset, progression, and resistance to cytotoxic drug therapy of many human solid tumors, including breast cancer [90].

As neural transcription factors, the ZIC family proteins (ZIC1–5) are said to play crucial roles in the development of the axial and limb bones as well as the proliferation of derivatives of the neural crest [91]. The focus of more recent study has been on the role of ZIC family proteins in the growth of malignancies [92]. It has been discovered that several ZIC family members express reversibly in various cancer types. ZIC1 expression was shown to be downregulated in 12 different pairings of breast cancer tissues, associated with a decrease in ZIC2 expression, according to research on the mRNA levels of the ZIC family members in breast cancer [93].