Colorectal cancer is one of the most common types of cancer worldwide, with a multifactorial digestive pathology. Evidence has suggested that gut microbial dysbiosis is connected to the development of colorectal cancer by generating cancer cell-conducive microenvironments. Nevertheless, the relationship between colorectal cancer pathogenesis and microorganisms has not been fully clarified to date. Here, we addressed this issue and determined the cancer-causing potential of the culture filtrate and proteins of Klebsiella pneumoniae on healthy cells. In this study, the culture filtrate and total proteins of K. pneumoniae isolated from patients with colorectal cancer were investigated to determine their cytotoxic effects against the normal human fibroblast PCS-201-012 cell model. As a result of the isolation procedure, three different K. pneumoniae strains (named Kp1, Kp2, and Kp3) were obtained from biopsy samples. Their 16S rRNA gene sequences were submitted to the GenBank database under the accession numbers MK156319, MK156320, and MK156321, respectively. The WST-8 and hemolysis tests were performed to examine the exacerbating effect of these strains on normal cells. The apoptosis-inducing ability of the isolated strains was characterized based on a combination of several techniques: determination of caspase-3 activity, JC-1 mitochondrial assay, and flow cytometry-based FITC-Annexin-V/PI double staining. Moreover, the expression profiles of four candidate genes (APC, SMAD, KRAS, TP53), which play important roles in the development of colorectal cancer, were analyzed by the qRT-PCR method. Cell biology experiments demonstrated that the culture filtrate and proteins of the related strains clearly cause cell death in normal human fibroblasts due to increased inflammatory response and necrosis. Furthermore, the culture filtrates and proteins led to a decrease in the expression of tumor suppressor genes TP53, SMAD, and APC and an increase in the expression of the KRAS oncogene, emphasizing the tumorigenicity of the strains in colorectal cancer. These results revealed that K. pneumoniae strains are capable of triggering cytotoxicity in normal human fibroblast cells.

Resistance to chemotherapy remains a major obstacle in effective cancer treatment. To address this challenge, we developed multifunctional Fe3O4@SiO2(FITC)-BTN/FA/3AB nanoparticles aimed at selectively enhancing the therapeutic efficacy of 3-aminobenzamide (3AB) while minimizing systemic toxicity, particularly when combined with low-dose cisplatin chemotherapy. In this study, we demonstrate that these nanoparticles not only exhibit potent cytotoxic effects against lung adenocarcinoma cells but also show significant therapeutic potential in human cervical adenocarcinoma models. We systematically evaluated their performance through cellular uptake assay, viability assay, apoptosis analysis, single-cell colony formation assay, mitochondrial membrane potential (MMP) assay, and quantitative PCR (qPCR). Our findings reveal that the nanoparticle formulation efficiently facilitates the intracellular delivery of 3AB, leading to robust inhibition of tumor cell proliferation and migration. Overall, the Fe3O4@SiO2(FITC)-BTN/FA/3AB nanoparticle system represents a promising platform for intratumoral therapy, offering a targeted strategy to potentiate the efficacy of PARP1 inhibition in DNA repair for cervical cancer treatment.

Chemotherapy is one of the main treatment methods for cancer patients, but its effectiveness is limited by drug resistance. Combining a chemotherapeutic agent with targeted molecular therapy may improve the curative effect of the chemotherapeutic agent. In this study, we investigate the efficacy of combining a 3-Aminobenzamide (3AB)-linked multifunctional platform with low-dose cisplatin chemotherapy aiming to modulate poly [ADP-ribose] polymerase 1 (PARP1) function in DNA repair to increase cytotoxic activity of the platinum-based cisplatin. The structure of the synthesized nanoplatforms was characterized by several physicochemical techniques, including dynamic light scattering (DLS), Fourier-transform infrared spectroscopy (FT-IR), scanning electron microscopy (SEM) and an in vitro pH-dependent release study. Cellular uptake experiments demonstrated preferentially targeted delivery of nanoparticles in lung carcinoma A549 cells, whereas the cellular uptake capacity was minimal in normal lung BEAS-2B cells. On the other hand, cytotoxicity experiments showed a reduction of cancer cell viability compared to free formulations. Furthermore, the combination treatment was examined by detecting the loss of mitochondrial membrane potential and the apoptotic cell population, confirming the treatment's functional involvement in apoptosis. Soft agar colony formation and cell invasion tests were also performed to detect the cancer cell's tumorigenic potential, confirming the synergistic effect of this combination in the reduction of tumorigenicity. Moreover, we analyzed the expression profiles of three candidate genes, which play important roles in cancer initiation, promotion and progression. Cell biology experiments indicated that this novel combination treatment possesses significant synergy between 3AB and low-dose cisplatin and is promising for development as an antitumor treatment for lung cancer.

In cancer treatments, many natural and synthetic products have been examined; among them, protease inhibitors are promising candidates for anti-cancer agents. Since dysregulated proteolytic activities can contribute to tumor development and metastasis, antagonization of proteases with tailored inhibitors is an encouraging approach. Although adverse effects of early designs of these inhibitors disappeared after the introduction of next-generation agents, most of the proposed inhibitors did not pass the early stages of clinical trials due to their nonspecific toxicity and lack of pharmacological effects. Therefore, new applications that modulate proteases more specifically and serve their programmed way of administration are highly appreciated. In this context, nanosized drug delivery systems have attracted much attention because preliminary studies have demonstrated that the therapeutic capacity of inhibitors has been improved significantly with encapsulated formulation as compared to their free forms. Here, we address this issue and discuss the current application and future clinical prospects of this potential combination towards targeted protease-based cancer therapy.

Today's nanoparticle technology enables the synthesis of nanoparticle-based drug delivery systems with desired size, shape, and materials especially for the applications of cancer nanomedicine. Thereby, understanding impact of particle sizes on anticancer activity will contribute to development of new drug delivery systems in cancer therapy. For this reason, in this study, two different sized nanoparticles (with -55 and 314 nm) were used as drug delivery systems and the effects of their size on the cellular uptake, cytotoxicity and apoptosis were investigated against the human colon carcinoma Caco-2 and HCT-116 cells. The results demonstrated that small nanoparticles promoted fast nanoparticle accumulation in both cancer cells in comparison to large particles. Small nanoparticles exhibited higher cytotoxicity in cancer cells with lower half maximal inhibitory concentration (IC50) values than large nanoparticles in 48 h. On the other hand, both nanoparticles showed similar IC50 values after 72 h prolonged exposure. Moreover, it was found that small nanoparticles increased the number of apoptotic cells in 24 h, whereas large nanoparticles induced apoptosis when exposure time increased to 72 h. These observations show that small sized drug delivery systems could be more efficient for improving the anticancer effects of chemotherapeutic drugs against human colon carcinoma as compared to large sized drug delivery systems.

İlaç nanotaşıyıcıları, kontrollü ve sürekli ilaç salım özellikleri ile kanser tedavisinde büyük bir potansiyele sahiptir. Bu nanotaşıyıcılar pasif veya aktif hedefli olarak ilaç taşınımı sağlayabilmektedir, ancak aktif hedefli muadillerine göre, pasif hedefli nanotaşıyıcılar tümörlü dokularda daha yavaş ve düşük düzeyde ilaç birikimi sağladığından kanserli hücrelerin yanında sağlıklı hücrelerde uzun süre bu nanotaşıyıcılara maruz kalmaktadır. Bu nedenle, bu çalışmada, pasif hedefli ilaç nanotaşıyıcıların insan akciğer epitel BEAS-2B hücreleri ve insan akciğer kanser A549 hücreleri üzerindeki uzun dönem etkileri araştırıldı. Bunun için, görüntüleme ve tedavi edici özellikleri bir arada barından Fe3O4@SiO2(FITC)-DOX formülasyonuna sahip ilaç nanotaşıyıcıları kullanıldı ve hücresel birikim, sitotoksisite ve apoptoz üzerindeki etkileri araştırıldı. Hücresel alım ve sitotoksisite deneyleri, pasif hedefli nanotaşıyıcıların kanser hücresi canlılığının etkin bir şekilde azalttığını gösterirken, 24 saatlik inkübasyon sürecinde sağlıklı hücreler üzerinde kayda değer bir etki görülmedi. Ancak 96 saatlik uzun inkübasyon sürecinde, sağlıklı BEAS-2B hücreleri makul seviyelerde nanotaşıyıcı alımı gerçekleştirirken, A549 kanser hücrelerine kıyasla düşük düzeylerde ilaç-aracılı sitotoksisite sergiledi. Ayrıca, nanotaşıyıcılar A549 hücrelerindeki apoptoz seviyelerini önemli ölçüde artırırken, BEAS-2B hücrelerinde 96 saat sonunda dahi apoptotik etki göstermedi. Bu sonuçlar, pasif hedefli inorganik ilaç nanotaşıyıcıların, sağlıklı hücreleri ihmal edilebilir düzeyde etkileyerek, antikanser ilaçların kemoterapötik etkilerini artırmada umut verici olduğunu göstermektedir.

Günümüz nanopartikül teknolojisi, özellikle kanser nanotıp uygulamalarında kullanılmak üzere istenilen boyut, şekil ve malzemeye sahip nanopartikül-temelli ilaç taşıyıcı sistemlerinin sentezine olanak sağlamaktadır. Dolayısıyla, partikül boyutlarının antikanser aktivite üzerindeki etkisini anlamak, kanser tedavisinde yeni ilaç taşıyıcı sistemlerin geliştirilmesine katkıda bulunacaktır. Bu nedenle, bu çalışmada, ilaç taşıyıcı sistemler olarak iki farklı büyüklükteki inorganik temelli nanopartiküller (~ 55 ve 314 nm) kullanıldı ve boyutlarının hücresel birikim, sitotoksisite ve apoptoz üzerindeki etkileri insan kolon kanseri Caco-2 ve HCT-116 hücrelerine karşı araştırıldı. Elde edilen sonuçlar, büyük nanopartiküllerle karşılaştırıldığında küçük nanopartiküllerin her iki kanser hücresinde de hızlı nanopartikül birikimini desteklediğini gösterdi. Küçük nanopartiküller, büyük nanopartiküllere göre 48 saat içinde daha düşük yarı-maksimum inhibisyon konsantrasyonu (IC50) değerleri ile kanser hücrelerinde daha yüksek sitotoksisite sergiledi. Öte yandan, her iki nanopartikül de 72 saate varan inkübasyon süreleri sonrası benzer IC50 değerleri gösterdi. Ayrıca, küçük nanopartiküller 24 saatte apoptotik hücrelerin sayısını artırırken, büyük nanopartiküllerin 72 saatlik süre içerisinde apoptozu indüklediği belirlendi. Bu gözlemler, küçük boyutlu ilaç taşıma sistemlerinin, büyük boyutlu ilaç taşıma sistemleri ile karşılaştırıldığında, insan kolon kanseri hücrelerinde kemoterapötik ilaçların antikanser etkilerini artırmada daha verimli olduğunu göstermektedir.

Cysteine cathepsins are lysosomal enzymes belonging to the papain family. Their expression is misregulated in a wide variety of tumors, and ample data prove their involvement in cancer progression, angiogenesis, metastasis, and in the occurrence of drug resistance. However, while their overexpression is usually associated with highly aggressive tumor phenotypes, their mechanistic role in cancer progression is still to be determined to develop new therapeutic strategies. In this review, we highlight the literature related to the role of the cysteine cathepsins in cancer biology, with particular emphasis on their input into tumor biology.

Chemotherapy frequently involves combination treatment protocols to maximize tumor cell killing. Unfortunately these intensive chemotherapeutic regimes, often show disappointing results due to the development of drug resistance and higher nonspecific toxicity on normal tissues. In cancer treatment, it is critically important to minimize toxicity while preserving efficacy. We have previously addressed this issue and proposed a nanoparticle-based combination therapy involving both a molecularly targeted therapy and chemotherapeutic agent for neutralizing antiapoptotic survivin (BIRC5) to potentiate the efficacy of doxorubicin (DOX). Although the particles exhibited strong anticancer effect on the lung carcinoma A549 and the cervical carcinoma HeLa cells, there were lower-level therapeutic outcomes on the colon carcinoma HCT-116, the leukemia Jurkat and the pancreatic carcinoma MIA PaCa-2 cells. Since targeted therapies are one of the key approaches for overcoming drug resistance, tailoring the treatment of cancer cells with distinct characteristics is necessary to improve the therapeutic outcome of cancer therapy and to minimize potential pharmacokinetic interactions of drugs. In the light of this issue, this study examined whether a cascade therapy with low-dose DOX and survivin-targeted tailored nanoparticles is more effective at sensitizing HCT-116, Jurkat and MIA PaCa-2 cancer cells to DOX-chemotherapy than simultaneous combination therapy. The results demonstrated that the sequential therapy with the protocol comprising addition of the nanoparticles after incubation of cells with DOX clearly advanced the therapeutic outcome of related cancer cells, whereas the reverse protocol resulted in a reduction or delay in apoptosis, emphasizing the critical importance of formulating synergistic drug combinations in cancer therapy.

Drug targeting and stimuli-responsive drug release are 2 active areas of cancer research and hold tremendous potential in the management of cancer drug resistance. In this study, I addressed this issue and focused on the synthesis and characterization of pH-responsive Fe3O4@SiO2(FITC)-BTN/folic acid/DOX multifunctional nanoparticles aiming to increase drug accumulation in malignancies with both dual active targeting and endosomal drug release properties. Dye-doped silica magnetic-fluorescent composite was constructed by a simple coprecipitation of Fe+2/Fe+3 salts followed by sol-gel formation and dual-targeting function was obtained by conjugating folate and biotin moieties on the silica surface of nanoparticles via an esterification reaction. Doxorubicin was then successfully attached on the amine-functionalized nanoparticles using a pH-sensitive Schiff-base formation. The physicochemical characterization of the structure was performed by dynamic light scattering, zeta potential measurement, X-ray diffraction, Fourier transform infrared spectroscopy, electron microscopy techniques, and an in vitro pH-dependent release study. Cellular uptake and cytotoxicity experiments demonstrated an enhanced intracellular delivery and reduction of cancer cell viability in the cervical carcinoma HeLa cell line. Furthermore, proapoptotic studies showed that the nanoparticles increased the apoptotic rates within the same cancer cells. The preliminary cell tests confirm the potential of these multifunctional nanoparticles against the development of drug resistance in cancer cells.

Nanopartikul-aracili ilac hedefleme kanser arastirmalarinin aktif bir alani olup, tumor dokusuna ozgun antikanser etkinligi artirmada cok onemli bir potansiyele sahiptir. Bu calismada, hedefleme verimlilik oranlarinin belirlenmesi icin pasif ve aktif hedefli nanopartikullerin tumor hedefleme kabiliyetleri, sirasiyla artmis gecirgenlik ve alikonma (EPR) etkisi ve biyotin reseptorlerini hedefleme yaklasimlari karsilastirilarak incelendi. Bunun icin, goruntuleme ve tedavi edici ozellikleri bir arada barindan Fe 3 O 4 @SiO 2 (FITC)-DOX (pasif hedefleme icin) ve Fe 3 O 4 @SiO 2 (FITC)-BTN/DOX (aktif hedefleme icin) multifonksiyonel nanopartikulleri kullanildi. Nanopartikullerin tumor hucrelerindeki birikiminin izlenmesi ve miktarsal olcumu icin floresan mikroskopu ve akim sitometresi kullanildi. Elde edilen sonuclar, pasif hedeflemeyle karsilastirildiginda aktif hedefleme stratejisinin servikal karsinoma HeLa hucrelerindeki nanopartikul birikimini 2 kat gibi onemli bir olcude artirdigini gosterdi. Aktif hedefli nanopartikuller, pasif hedefli nanopartikulerden yaklasik 2.5 kat daha dusuk yari-maksimum inhibisyon konsantrasyonu (IC 50 ) degeri ile kanser hucrelerinde daha yuksek sitotoksisite sergiledi. Ayrica, pasif hedefli nanopartikullerle karsilastirildiginda, aktif hedefli nanopartikullerin apoptotik hucre sayisini yaklasik % 21.1 oraninda artirdigi bulundu. Bu gozlemler, aktif tumor hedefli ilac tasima sistemlerinin, pasif tumor hedefli ilac tasima sistemleri ile karsilastirildiginda, antikanser ilaclarin kemoterapotik etkilerini artirmada daha umut verici oldugunu gostermektedir.

Classical chemotherapy uses chemotherapeutic agents as a mainstay of anticancer treatment. However, the development of multidrug resistance to chemotherapy limits the effectiveness of current cancer treatment. Nanosized bioconjugates combining a chemotherapeutic agent with a pharmacological approach may improve the curative effect of chemotherapeutic agents. Herein I addressed this issue by describing the synthesis, and testing of, pH-responsive Fe3O4@SiO2(FITC)-BTN/QUR/DOX multifunctional nanoparticles. The particles were designed to modulate resistance-mediating factors and to potentiate the efficacy of DOX against chemoresistance. The physicochemical properties of the nanoparticles were characterized based on the combination of several techniques: dynamic light scattering (DLS), zeta-potential measurement, Fourier transform infrared spectroscopy (FTIR), electron microscopy techniques (SEM and STEM with EDX) and an in vitro pH-dependent release study. Cellular uptake and cytotoxicity experiments demonstrated enhanced intracellular delivery and retention of nanoparticles in the cytoplasm and efficient reduction of cancer cell viability in drug-resistant lung carcinoma A549/DOX cell lines. This did not affect internalization and viability of an immortalized human lung epithelial cell line BEAS-2B. Moreover, proapoptotic and antiproliferative studies showed that Fe3O4@SiO2(FITC)-BTN/QUR/DOX nanoparticles can promote apoptosis, inhibit tumor cell proliferation, and enhance the chemotherapeutic effects of DOX against multidrug resistance. These results confirm that this multifunctional platform possesses significant synergy between QUR and DOX and is promising for development as an antitumor treatment in cancer therapy.

The yeast Pichia pastoris  expression system was investigated for the production of human cathepsin C (CatC) recombinant protein. The full-length CatC cDNA, corresponding to amino acids 12-475, was synthesized from interleukin-2 (IL-2) stimulated human peripheral blood mononuclear cells and subcloned in the pGEM-T cloning vector. After confirming the DNA sequence of the insert, the gene was cloned into the pPICZαA expression vector under the control of the methanol-inducible alcohol oxidase (AOX1) promoter and transformed to P. pastoris X-33 cells. The expressed protein was secreted into the culture medium through the α-factor mating signal sequence of the expression vector. Analysis of the culture supernatant revealed that the recombinant human CatC was secreted as a 58-kDa molecule, indicating that human CatC was accumulated in the culture supernatant as proform composed of the residual propart, the activation peptide, and the heavy and light chains. Extracellular recombinant proCatC was further activated by cysteine endoprotease papain in vitro and its activity was confirmed by assays using a synthetic substrate.

Purpose Resistance to chemotherapy is one of the major problems facing current cancer research. Enhancing tumor cell response to anticancer agents increases chemotherapeutic effectiveness. We have recently addressed this issue and reported on producing multifunctional nanoparticles (Fe 3 O 4 @SiO 2 (FITC)-FA/AICAR/DOX) aiming to overcome chemoresistance with synergetic effect of AICAR and DOX. In the present study, we demonstrated that these nanoparticles not only show enhanced cellular uptake and cytotoxic effect but can also show enhanced pro-apoptotic and anti-proliferative effects in five different tumor-derived cell lines (A549, HCT-116, HeLa, Jurkat and MIA PaCa-2). Methods The nanoparticles were examined by using flow cytometric analyses of apoptosis and cell cycle. In addition, we performed caspase-3 activity assay, which supported our flow cytometric data. Furthermore, we demonstrated the applicability of this approach in a variety of cancer types confirming the potential widespread utility of this approach. Results With the concept of co-delivery of AICAR and DOX in the nanoparticle formulation, the use of AICAR against survivin ( BIRC5 ) sensitized cancer cells to DOX chemotherapy which resulted in effective cancer cell elimination. These result showed that combination therapy involving both a molecularly targeted therapy and chemotherapeutic agent has the ability to retain and enhance therapeutic efficacy. Conclusion Fe 3 O 4 @SiO 2 (FITC)-FA/AICAR/DOX nanoparticles is superior to monotherapy via the synergetic effect of AICAR and DOX and also the nanoparticle formulation could overcome issues of toxicity with targeted therapy while maintaining the potent anticancer effects of AICAR and DOX. Graphical Abstract Apoptosis analysis of A549 cells by flow cytometry-based PE-annexin-V / 7-ADD double staining treated with low-dose (10 μg/ml) concentration of (1) Fe3O4@SiO2(FITC)-FA (2) Fe3O4@SiO2(FITC)-FA/AICAR, (3) Fe3O4@SiO2(FITC)-FA/DOX or (4) Fe3O4@SiO2(FITC)-FA/AICAR/DOX nanoparticles. Viable cells labelled with PE-annexin-V(-)/7-ADD(-), early apoptotic cells labelled with PE-annexin-V(+)/7-ADD(-) and apoptotic cells labelled with PE-annexin-V(+)/ 7-ADD(+) in flow cytometric graphics.

The success of cancer treatment depends on the response to chemotherapeutic agents. However, malignancies often acquire resistance to drugs if they are used frequently. Combination therapy involving both a chemotherapeutic agent and molecularly targeted therapy may have the ability to retain and enhance therapeutic efficacy. Here, we addressed this issue by examining the efficacy of a novel therapeutic strategy that combines AICAR and DOX within a multifunctional platform. In this context, we reported the bottom-up synthesis of Fe3O4@SiO2(FITC)-FA/AICAR/DOX multifunctional nanoparticles aiming to neutralize survivin (BIRC5) to potentiate the efficacy of DOX against chemoresistance. The structure of nanoparticles was characterized by dynamic light scattering (DLS), zeta-potential measurement, X-ray diffraction (XRD), Fourier transform infrared spectroscopy (FT-IR), thermogravimetric analysis (TGA), and electron microscopy (SEM and STEM with EDX) techniques. Cellular uptake and cytotoxicity experiments demonstrated preferentially targeted delivery of nanoparticles and an efficient reduction of cancer cell viability in five different tumor-derived cell lines (A549, HCT-116, HeLa, Jurkat, and MIA PaCa-2). These results indicate that the multifunctional nanoparticle system possesses high inhibitory drug association and sustained cytotoxic effect with good biocompatibility. This novel approach which combines AICAR and DOX within a single platform might be promising as an antitumor treatment for cancer.

Trypsin was immobilized by covalent binding to glutaraldehyde-activated silica with and without a spacer arm; 1,6-diaminohexane and polyethyleneglycol as well. The addition of polyethyleneglycol (PEG) to the immobilization media increased the activity of immobilized trypsin in organic solvents, whilst free trypsin activity disappeared under the same conditions. Thermal, pH, storage, and operational stabilities of the free and immobilized enzyme were found to be better than the free enzyme. Furthermore, use of immobilized enzyme for protein fragmentation was achieved by solid-phase, on-line, protein digestion in organic solvents. Reaction times were reduced to a few minutes and the sample handling was minimized.