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Comparison of the ability of 5-FU, 5-FU + ACF and 5-FU + irinotecan to inhibit the viability of (A) SW480, (B) HCT116 and (C) LS174T cell lines. Cells were treated with different concentrations of 5-FU as a single agent for 48 h, or pretreated with the 30% maximal inhibitory concentration of ACF or irinotecan for 24 h and then exposed to different concentration of 5-FU for 48 h. Cell viability was determined using an MTT assay. Results are the mean ± standard deviation of three experiments, each performed in triplicate. 5-FU, 5-fluorouracil; ACF, acriflavine.
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5-Fluorouracil (5-FU)-based chemotherapy improves the overall survival rates of patients with colorectal cancer (CRC). However, only a small proportion of patients respond to 5-FU when used as a single agent. The aim of the present study was to investigate whether the anticancer property of 5-FU is potentiated by combination treatment with acriflav...
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... of ACF and 5-FU co-treatment on the sensitivity of CRC cells to 5-FU. To assess the cytotoxic effects of ACF or 5-FU, CRC cells (SW480, HCT116 and LS174T) were treated with a graded concentration of drugs for 72 h and cell viability was determined using an MTT assay. The IC 50 values of ACF for SW480, HCT116 and LS174T cells were 0.75±0.10, 0.57±0.22 and 0.36±0.05 µM, respectively. ACF caused inhibitory effects on the cell growth in a dose-dependent manner (Fig. 1). The same pattern was also obtained for 5-FU (data not shown). The IC 30 values of ACF for SW480, HCT116 and LS174T cells were 0.36±0.07, 0.29±0.14 and 0.20±0.04 µM, respectively. The sensitivity of CRC cells against ACF and 5-FU was deter- mined by calculation of the IC 50 values as presented in Table II. SW480 cells exhibited the highest resistance to ACF and 5-FU in comparison with the other two cell lines. To determine the effect of ACF on the sensitivity of cells against 5-FU, cells were simultaneously treated with the IC 30 , the low cytotoxic concentration, of ACF and different concentrations of 5-FU for 72 h. As indicated in Table II, the results indicated that the co-treatment protocol was not able to significantly alter the IC 50 value of 5-FU on CRC cells. Therefore, an alternative treatment protocol was designed (pretreatment protocol). Fig. 2, the pattern of CRC cell responses to the cytotoxic effect of ACF + 5-FU is presented. In all ACF-pretreated cell lines, at the low concentration of 5-FU, an increased amount of cell death occurred. Furthermore, the IC 50 value of 5-FU in the pretreat- ment protocol was significantly lower compared with the IC 50 value of 5-FU when used as a single drug (Table IV). In fact, ACF pretreatment was able to sensitize CRC cells to the low concentration of ...
Context 2
... of ACF and 5-FU co-treatment on the sensitivity of CRC cells to 5-FU. To assess the cytotoxic effects of ACF or 5-FU, CRC cells (SW480, HCT116 and LS174T) were treated with a graded concentration of drugs for 72 h and cell viability was determined using an MTT assay. The IC 50 values of ACF for SW480, HCT116 and LS174T cells were 0.75±0.10, 0.57±0.22 and 0.36±0.05 µM, respectively. ACF caused inhibitory effects on the cell growth in a dose-dependent manner (Fig. 1). The same pattern was also obtained for 5-FU (data not shown). The IC 30 values of ACF for SW480, HCT116 and LS174T cells were 0.36±0.07, 0.29±0.14 and 0.20±0.04 µM, respectively. The sensitivity of CRC cells against ACF and 5-FU was deter- mined by calculation of the IC 50 values as presented in Table II. SW480 cells exhibited the highest resistance to ACF and 5-FU in comparison with the other two cell lines. To determine the effect of ACF on the sensitivity of cells against 5-FU, cells were simultaneously treated with the IC 30 , the low cytotoxic concentration, of ACF and different concentrations of 5-FU for 72 h. As indicated in Table II, the results indicated that the co-treatment protocol was not able to significantly alter the IC 50 value of 5-FU on CRC cells. Therefore, an alternative treatment protocol was designed (pretreatment protocol). Fig. 2, the pattern of CRC cell responses to the cytotoxic effect of ACF + 5-FU is presented. In all ACF-pretreated cell lines, at the low concentration of 5-FU, an increased amount of cell death occurred. Furthermore, the IC 50 value of 5-FU in the pretreat- ment protocol was significantly lower compared with the IC 50 value of 5-FU when used as a single drug (Table IV). In fact, ACF pretreatment was able to sensitize CRC cells to the low concentration of ...
Citations
... Different 9Acr doses and treatment periods led to downregulation of protein expression even at 4-8 h post-treatment time with a threshold at 5 µM 9Acr [37], which corresponds to our cytotoxicity findings. Similarly, acriflavine may also be incorporated along with 5-Fluorouracil chemotherapeutics for colorectal carcinoma treatment to boost efficiency [38]; either alone or combined, these compounds require 7.4-11.6 µM concentration to reach IC50 value. ...
Acridine and its derivatives (9-chloroacridine and 9-aminoacridine) are investigated here, supported on FAU type zeolite Y, as a delivery system of anticancer agents. FTIR/Raman spectros-copy and electron microscopy revealed successful drug loading on the zeolite surface, while spec-trofluorimetry was employed for drug quantification. The effects of the tested compounds on cell viability were evaluated using in vitro methylthiazol-tetrazolium (MTT) colorimetric technique against human colorectal carcinoma (cell line HCT-116) and MRC-5 fibroblasts. Zeolite structure remained unchanged during homogeneous drug impregnation with achieved drug loadings in the 18-21 mg/g range. The highest drug release, in the µM concentration range, with favourable kinetics was established for zeolite-supported 9-aminoacridine. The acridine delivery via zeolite carrier is viewed in terms of solvation energy and zeolite adsorption sites. The cytotoxic effect of supported acridines on HCT-116 cells reveals that the zeolite carrier improves toxicity, while the highest efficiency is displayed by zeolite-impregnated 9-aminoacridine. The 9-aminoacridine delivery via zeo-lite carrier favours healthy tissue preservation while accompanying increased toxicity toward cancer cells. Cytotoxicity results are well correlated with theoretical modelling and release study, providing promising results for applicative purposes.
... 75 ACF enhances the antitumor activity of sunitinib in a breast cancer model 78 and of 5-fluorouracil used in the treatment of colorectal cancer much better than irinotecan. 79 It acts on HIF-1 by reducing the expression of LOX and LOXL proteins (responsible for metastases), destroying the metastatic niches of breast cancer. 80 It was also proven that the synergistic effect of ACF and ABT-263 drugs strongly exerted triple negative breast cancer (TNBC) apoptosis. ...
Acriflavine (ACF) has been known for years as an antibacterial drug. The identification of key oncogenic mechanisms has brought, in recent years, a significant increase in studies on ACF as a multipurpose drug that would improve the prognosis for cancer patients. ACF interferes with the expression of the hypoxia inducible factor, thus acting on metastatic niches of tumors and significantly enhancing the effects of other anticancer therapies. It has been recognized as the most potent HIF-1 inhibitor out of the 336 drugs approved by the FDA. This work presents up-to-date knowledge about the mechanisms of action of ACF and its related prodrug systems in the context of anticancer and SARS-CoV-2 inhibitory properties. It explains the multitask nature of this drug and suggests mechanisms of ACF's action on the coronavirus. Other recent reports on ACF-based systems as potential antibacterial and antiviral drugs are also described.
... Highthroughput molecular screening revealed that ACF is an antitumor agent in colon cancer [24]. Some studies have shown that ACF can enhance the toxicity of chemotherapeutic compounds to cancer cells [25,26]. ACF is also commonly used as an inhibitor of HIF protein expression, which promotes the progression of various solid tumors and leukemia [27]. ...
Colon cancer ranks third worldwide, and it has a growing incidence with urbanization and industrialization. Drug resistance in colon cancer is gradually affecting the treatment. This study focused on the mechanisms by which acriflavine (ACF) enhances the radiosensitivity of colon cancer cells. First, the expression and activation levels of tumor suppressor protein p53 were shown high in normal cells and tissues in its detection, which suggests that p53 is likely to be a key factor in colon cancer. Then, the expression of p53 ended up increasing in ACF group after SW620 cells were cultured with ACF. In addition, ACF group had some other changes. The expression of mitochondrial related antiapoptotic protein Bcl-2 increased, while the expression of proapoptotic protein Bax, Bad, cytopigment C, and apoptotic inducer AIF decreased. At the same time, the ability of apoptosis was enhanced, and the ability of proliferation and invasion was decreased. This suggests that ACF can promote p53 expression and affect mitochondrial function and the radiosensitivity of SW620. The luciferase reporting experiment showed that there was a binding site between ACF and p53. Besides, when IR treatment was applied to SW620 with high p53 expression, there was an increase in the expression of Bcl-2 in SW620 and decrease in Bax, Bad, and cytopigment C in AIF. Meanwhile, the cell apoptosis became stronger, and the proliferation and invasion became weaker. The experimental results were similar to those of SW620 cells cultured with ACF, suggesting that p53 is an intermediate factor in the regulation of SW620 by ACF. Finally, in this study, cells were cultured with ACF, and p53 was knocked down at the same time. The experimental results showed that after p53 was knocked down. ACF’s ability to regulate SW620 is partially removed. This confirms the view that ACF regulates SW620 cells by regulating p53. In summary, this study found the mechanism by which ACF causes mitochondrial dysfunction and improves the radiosensitivity of colon cancer cells by activating the tumor suppressor protein p53, which may contribute to solving the drug resistance in colon cancer.
... Low curcumin doses enhance 5-FU effects against colorectal cancer cells and dramatically reduce the number and volume of tumors in mice subjected to the colitisassociated CRC protocol [174][175][176][177]. Combined 5-FU with thymoquinone in sublethal doses was effective against CD133 + tumor stem cells and showed no toxic effects on healthy organoids [178]. Additionally, many other naturally occurring small molecules used as Wnt inhibitors have had promising effects when combined with chemotherapeutic agents, the most observed effects being inhibition of proliferation and induction of apoptosis [179][180][181][182][183][184][185][186]. Therefore, combining Wnt inhibitor drugs with current chemotherapeutic treatment might be an interesting approach to overcome CRC carcinogenesis. ...
Simple Summary
Colorectal cancer (CRC) is an emerging public health problem and the second leading cause of death worldwide, with a significant socioeconomic impact in several countries. The 5-year survival rate is only 12% due to the lack of early diagnosis and resistance to available treatments, and the canonical Wnt signaling pathway is involved in this process. This review underlines the importance of understanding the fundamental roles of this pathway in physiological and pathological contexts and analyzes the use of naturally occurring small molecules that inhibits the Wnt/β-catenin pathway in experimental models of CRC. We also discuss the progress and challenges of moving these small molecules off the laboratory bench into the clinical platform.
Abstract
Colorectal cancer (CRC) ranks second in the number of cancer deaths worldwide, mainly due to late diagnoses, which restrict treatment in the potentially curable stages and decrease patient survival. The treatment of CRC involves surgery to remove the tumor tissue, in addition to radiotherapy and systemic chemotherapy sessions. However, almost half of patients are resistant to these treatments, especially in metastatic cases, where the 5-year survival rate is only 12%. This factor may be related to the intratumoral heterogeneity, tumor microenvironment (TME), and the presence of cancer stem cells (CSCs), which is impossible to resolve with the standard approaches currently available in clinical practice. CSCs are APC-deficient, and the search for alternative therapeutic agents such as small molecules from natural sources is a promising strategy, as these substances have several antitumor properties. Many of those interfere with the regulation of signaling pathways at the central core of CRC development, such as the Wnt/β-catenin, which plays a crucial role in the cell proliferation and stemness in the tumor. This review will discuss the use of naturally occurring small molecules inhibiting the Wnt/β-catenin pathway in experimental CRC models over the past decade, highlighting the molecular targets in the Wnt/β-catenin pathway and the mechanisms through which these molecules perform their antitumor activities.
... These reports have also shown that the systematic administration of ACF decreased tumor size during the first two weeks, showing the promising use of ACF in glioma therapy [55]. Interestingly, ACF increased the cytotoxic effect of 5-fluorouracil (5-FU) in colorectal cancer cell lines, suggesting the development of new chemotherapy regimens that administer ACF in 5-FU-resistant patients with advanced colorectal cancer [56]. The pretreatment of human perihilar cholangiocarcinoma cells (SK-ChA-1) with ACF enhanced photodynamic therapy (PDT) efficacy through the inhibition of HIF-1 and topoisomerases I and II [57]. ...
Drug repurposing has lately received increasing interest in several diseases especially in cancers due to its advantages in facilitating the development of new therapeutic strategies, by adopting a cost-friendly approach and avoiding the strict Food and Drug Administration (FDA) regulations. Acriflavine (ACF) is an FDA approved molecule that has been extensively studied since 1912 with antiseptic, trypanocidal, anti-viral, anti-bacterial and anti-cancer effects. ACF has been shown to block the growth of solid and hematopoietic tumor cells. Indeed, ACF acts as an inhibitor of various proteins including DNA-dependent protein kinases C (DNA-PKcs), topoisomerase I and II, hypoxia-inducible factor 1α (HIF-1α), in addition to its recent discovery as an inhibitor of the signal transducer and activator of transcription (STAT). Chronic myeloid leukemia (CML) is a clonal myeloproliferative disorder characterized by the expression of the constitutively active tyrosine kinase BCR-ABL. This protein allows the activation of several signaling pathways known for their role in cell proliferation and survival such as JAK/STAT pathway. CML therapy, based on tyrosine kinase inhibitors (TKIs), such as imatinib (IM), is highly effective. However, 15% of patients are refractory to IM, where in some cases 20-30% of patients become resistant. Thus, we suggest the repurposing of ACF in CML after IM failure or in combination with IM to improve the antitumor effects of IM. In this review, we present the different pharmacological properties of ACF along with its anti-leukemic effects in the hope of its repurposing in CML therapy.
... [3][4][5][6][7] Moreover, ACF has also increased the cytotoxic effects of chemotherapeutic compounds such as 5-fluorouracil or melphalan in cancer cells. 8,9 Other studies have demonstrated that ACF effectively inhibits drug resistance and epithelial-mesenchymal transition (EMT) of pancreatic and hepatic cancer cells. 7 Importantly, long-term administration of ACF to AIDS patients, as an antiviral agent, has not revealed any major side effects suggesting that it could be employed alone or in combination with other drugs to overcome chemotherapy resistance of cancer cells. ...
Acriflavine (ACF) is an antiseptic with anticancer properties, blocking the growth of solid and haematopoietic tumour cells. Moreover, this compound has been also shown to overcome the resistance of cancer cells to chemotherapeutic agents. ACF has been shown to target hypoxia‐inducible factors (HIFs) activity, which are key effectors of hypoxia‐mediated chemoresistance. In this study, we showed that ACF inhibits the growth and survival of chronic myeloid leukaemia (CML) and acute myeloid leukaemia (AML) cell lines in normoxic conditions. We further demonstrated that ACF down‐regulates STAT5 expression in CML and AML cells but activates STAT3 in CML cells in a HIF‐independent manner. In addition, we demonstrated that ACF suppresses the resistance of CML cells to tyrosine kinase inhibitors, such as imatinib. Our data suggest that the dual effect of ACF might be exploited to eradicate de novo or acquired resistance of myeloid leukaemia cells to chemotherapy.
... For this reason, a co-encapsulation was not intended, and monotherapy was further investigated. ACF has been shown to sensitize CRC cells to 5-FU in pretreatment but not in combination (Zargar et al., 2018). Therefore, combinations with acriflavine may associate cytotoxic agents, but with a focus on the schedule of each drug administration and of the cytotoxic drug choice. ...
Crosstalk between cancer-associated fibroblasts (CAFs) and colorectal cancer cells promotes tumor growth and contributes to chemoresistance. In this study, we assessed the sensitivity of a primary CAF cell line, CT5.3hTERT, to standard-of-care and alternative cytotoxic treatments. Paclitaxel (PTX) and acriflavine (ACF) were identified as the most promising molecules to inhibit CAF development. To allow the translational use of both drugs, we developed lipid nanocapsule (LNC) formulations for PTX and ACF. Finally, we mixed CAFs and tumor cell lines in a cocultured spheroid, and the effect of both drugs was investigated by histological analyses. We demonstrated CAF inhibition by LNC-ACF and whole tumor inhibition by LNC-PTX. Altogether, we proposed a new strategy to reduce CAF populations in the colorectal microenvironment that should be tested in vivo.
... Cancer is one of the leading causes of death worldwide, creating a global health problem [1] . The incidence of colorectal cancer (CRC) ranks third, it is among the most commonly diagnosed cancers, and the second leading cause of mortality [2] . More than 700000 people die annually due to CRC [3] . ...
Colorectal cancer (CRC) has been designated a major global problem, especially due to its high prevalence in developed countries. CRC mostly occurs sporadically (75%-80%), and only 20%-25% of patients have a family history. Several processes are involved in the development of CRC such as a combination of genetic and epigenetic alterations. Epigenetic changes, including DNA methylation play a vital role in the progression of CRC. Complex interactions between susceptibility genes and environmental factors, such as a diet and sedentary lifestyle, lead to the development of CRC. Clinical and experimental studies have confirmed the beneficial effects of dietary polyunsaturated fatty acids (PUFAs) in preventing CRC. From a mechanistic viewpoint, it has been suggested that PUFAs are pleiotropic agents that alter chromatin remodeling, membrane structure and downstream cell signaling. Moreover, PUFAs can alter the epigenome via modulation of DNA methylation. In this review, we summarize recent investigations linking PUFAs and DNA methylation-associated CRC risk.
Cancer is a global health problem being the second worldwide cause of deaths right after cardiovascular diseases. The main methods of cancer treatment involve surgery, radiation and chemotherapy with an emphasis on the latter. Thus development of nanochemistry and nanomedicine in a search for more effective and safer cancer treatment is an important area of current research. Below, we present interaction of doxorubicin and acriflavine and the cytotoxicity of these drug nano-complexes towards cervical cancer (HeLa) cells. Experimental results obtained from NMR measurements and fluorescence spectroscopy show that the drugs' interaction was due to van der Waals forces, formation of hydrogen bonds and π-π stacking. Quantum molecular simulations confirmed the experimental results with regard to existing π-π stacking. Additionally it was shown that, at the level of theory applied (DFT, triple zeta basis set), the stacking interactions comprise the most preferable interactions (the lowest ΔG ca. -12 kcal mol-1) both between the molecules forming the acriflavine system and between the other component - another drug (doxorubicin) dimer. Biological tests performed on HeLa cells showed high cytotoxicity of the complexes, comparable to free drugs (ACF and DOX), both after 24 and 48 hours of incubation. For non-cancerous cells, a statistically significant difference in the cytotoxicity of drugs and complexes was observed in the case of a short incubation period. The results of the uptake study showed significantly more efficient cellular uptake of acriflavine than doxorubicin, whether administered alone or in combination with an anthracycline. The mechanism determining the selective uptake of acriflavine and ACF : DOX complexes towards non-cancer and cancer cells should be better understood in the future, as it may be of key importance in the design of complexes with toxic anti-cancer drugs.
Despite the multidisciplinary standard treatment of glioblastoma (GB) consisting of maximal surgical resection, followed by radiotherapy (RT) plus concomitant chemotherapy with temozolomide (TMZ), the majority of patients experience tumor progression and almost universal mortality. In recent years, efforts have been made to create new agents for GB treatment, of which azo-dyes proved to be potential candidates, showing antiproliferative effects by inducing apoptosis and by inhibiting different signaling pathways. In this study we evaluated the antiproliferative the effect of six azo-dyes and TMZ on a low passage human GB cell line using MTT assay. We found that all compounds proved antiproliferative properties on GB cells. At equimolar concentrations azo-dyes induced more cytotoxic effect than TMZ. We found that Methyl Orange required the lowest IC50 for 3 days of treatment (26.4684 μM), whilst for 7 days of treatment, two azo dyes proved to have the highest potency: Methyl Orange IC50 = 13.8808 μM and Sudan I IC50 = 12.4829 μM. The highest IC50 was determined for TMZ under both experimental situations. Conclusions: Our research represents a novelty, by offering unique valuable data regarding the azo-dye cyototoxic effects in high grade brain tumors. This study may focus the attention on azo-dye agents that may represent an insufficient exploited source of agents for cancer treatment.
