Normal Healthy Cells Research Articles

Downregulation of miR-17 suppresses TGF-β1-mediated renal fibrosis through targeting Smad7.

MiR-17 is found upregulated in diabetic mice; however, its effect(s) on renal fibrosis of diabetic nephropathy remain(s) unknown. This study aimed to explore the mechanism underlying the downregulation of miR-17 in renal fibrosis of diabetic nephropathy (DN). Patients with diabetes mellitus (DM) and DN and normal healthy individual controls, mice (db/db, db/m), and human mesangial cells (HMCs) and human proximal tubule epithelial cells (HK-2) were used as research subjects in the study. Quantitative real-time polymerase chain reaction (qRT-PCR) was performed to measure the expression of miR-17 in the serum samples, renal tissues and cells. Acid-Schiff (PAS) and Masson staining experiments were performed to detect glomerular mesangial matrix and collagen deposition. Levels of fibrosis-related proteins (E-Cadherin (E-cad), vimentin, fibronectin and collagen I) were measured by Western blot (WB). The target gene of miR-17 was predicted by TargetScan 7.2 and confirmed by dual-luciferase reporter analysis. The study found that miR-17 expression was elevated in the serums of DN patients as well as in the serums and kidney tissues of db/db mice. db/db mice showed a severe renal fibrosis condition. The levels of E-cad in db/db mice, HMC and HK-2 cells were increased by downregulating miR-17 expression, while expressions of vimentin, fibronectin and collagen I were reduced. Smad7 was predicted to be the target gene of miR-17, and its expression was promoted by downregulation of miR-17. Moreover, the reduced Smad7 expression could inhibit the expressions of fibrosis-related proteins, which, however, can be ameliorated by the downregulation of miR-17. In addition, downregulation of miR-17 could suppress renal fibrosis mediated by TGF-β1 through targeting Smad7, which might be a clinical therapeutic target for patients with DN.

Novel Pluronic F‐127‐coated ZnO nanoparticles: Synthesis, characterization, and their in‐vitro cytotoxicity evaluation

AbstractZinc oxide (ZnO) nanoparticles (NPs) are increasingly recognized for their potential use in various biomedical applications. However, the tendency of bare ZnO NPs to aggregate and their poor stability in the biological environment reduce their biocompatibility and lead to cytotoxicity in normal healthy cells. This study attempted to avoid this by synthesizing a novel Pluronic F‐127‐coated ZnO NPs and evaluating their cytotoxicity impact on normal healthy human endothelial AE.hy296 cells. The characterization results revealed that Pluronic coating process remarkably enhanced the stability of ZnO NPs and did not alter their desired crystalline structure. The cytotoxicity of bare and Pluronic F‐127‐coated ZnO NPs were evaluated through MTT assay and microscopy imaging. The results of the cytotoxicity tests showed that Pluronic F‐127 modification significantly improved the biocompatibility and reduced the cytotoxicity of ZnO NPs toward the normal endothelial human EA.hy926 cells. Based on their high crystallinity, stability, and biocompatibility, Pluronic F‐123‐coated ZnO NPs hold promising potential for miscellaneous biomedical applications.

FOXD3‑AS1/miR‑128‑3p/LIMK1 axis regulates cervical cancer progression

Long non-coding RNA forkhead box D3 antisense RNA 1 (FOXD3-AS1) functions as an oncogenic regulator in several types of cancer, including breast cancer, glioma and cervical cancer. However, the effects and mechanisms underlying FOXD3-AS1 in cervical cancer (CC) are not completely understood. The present study aimed to investigate the biological functions and potential molecular mechanisms underlying FOXD3-AS1 in CC progression. Reverse transcription-quantitative PCR was performed to detect FOXD3-AS1, microRNA (miR)-128-3p and LIM domain kinase 1 (LIMK1) expression levels in CC tissues and cells. Immunohistochemical staining and western blotting were conducted to assess LIMK1 protein expression levels in CC tissues and cells, respectively. Cell Counting Kit-8 and BrdU assays were used to determine the role of FOXD3-AS1 in regulating cell proliferation. CC cell migration and invasion were assessed by performing Transwell assays. Dual-luciferase reporter assays were conducted to verify the binding between miR-128-3p and FOXD3-AS1. FOXD3-AS1 expression was significantly increased in CC tissues and cell lines compared with adjacent healthy tissues and normal cervical epithelial cells, respectively. High FOXD3-AS1 expression was significantly associated with poor differentiation of tumor tissues, increased tumor size and positive lymph node metastasis. FOXD3-AS1 overexpression significantly increased CC cell proliferation, migration and invasion compared with the negative control (NC) group, whereas FOXD3-AS1 knockdown resulted in the opposite effects compared with the small interfering RNA-NC group. Moreover, the results demonstrated that FOXD3-AS1 targeted and negatively regulated miR-128-3p, which indirectly upregulated LIMK1 expression. Therefore, the present study demonstrated that FOXD3-AS1 upregulated LIMK1 expression via competitively sponging miR-128-3p in CC cells, promoting CC progression.

Essentially new inhibitors of metastasis of malignant tumors for chemotherapy-sparing treatment

Georgia has received a patent for the first medicine (Amphicezine) of this class, which does not damage the normal healthy cells in the body. There are presented the synthesis schemes and methods of prospective metastasis inhibitors by the authors.

A theoretical study to focus a polychromatic synchrotron X-ray beam for microbeam radiation therapy

Microbeam radiotherapy (MRT) appears to target tumour cells while sparing normal healthy cells. However, it is in its preclinical stage at present. Clinical trials of MRT can be done in the Imaging and Medical Beamline (IMBL) of the Australian Synchrotron provided that the dose rate deliverable by the X-ray beam in Hutch 3B of the IMBL, where the required patient positioning equipment is located, is sufficiently high for MRT. At present the deliverable dose rate there is only 75Gys−1. However, a deliverable dose rate of between and including 375Gys−1 and 750Gys−1 is required for doing MRT clinical trials in Hutch 3B due to the irradiation profile inherent to MRT. The height of the focused beam in Hutch 3B must also be between and including 1.0 mm and 10 mm. A kinoform compound refractive lens (kinoform CRL) made of [3.5×1018Patomscm−3]-doped Si is proposed to be used in the Scenario where the X-ray beam, produced by a superconducting multipole wiggler source (SCMPW) operating at a maximum magnetic field of 4.000 T, is filtered by 0.6 mm of diamond before reaching the kinoform CRL. This is the Diamond-Optic Scenario. In this Scenario, the dose rate deliverable in Hutch 3B using the optic ranges from 380Gys−1 to 1300Gys−1 with beam heights ranging from 3.55 mm down to 1.0 mm, respectively. Also, the weighted average photon energies of the beam in Hutch 3B in this Scenario are almost entirely suitable for MRT in the IMBL. The X-ray beam in this Scenario can heat the P-doped Si optic to a few hundred degrees Kelvin above the hypothesised melting point of the P-doped Si due to its high power density. Therefore, the optic must be cooled lest it melt. The Diamond-Optic Scenario is not used routinely in the IMBL due to its excessive power. A variation of the Diamond-Optic Scenario, namely that the kinoform CRL be placed before sensitive components instead of after them is proposed for the routine use of this Scenario.

Drug delivery and in vitro biological effects of gum ghatti-modified hydroxyapatite nanoporous composites

The present study investigates the influence of gum ghatti (GG) concentration on the porosity of hydroxyapatite (HAP), where the final composite has been tested for the effects of drug delivery, antibacterial activity, and in vitro cytotoxicity. Following the synthesis of GG-HAP composite by a simple chemical precipitation method, the various analyses were employed to understand the crystallinity, morphology, chemical composition, surface functionality and internal bonding, etc. From the comparison of physical properties among the sintered and un-sintered GG-HAP composites and that too with different concentrations of GG (1%, 5%, and 10%), we observed the superior properties for the samples formed from higher GG concentration (10%) with sintering. We observed the lowest crystal size of 30 nm with 95% crystallinity, a surface area of 108 m2/g, a total pore volume of 0.2 mL/g, an average pore diameter of 33 Å for the Sin-10% GG-HAP composite. In addition, the biocompatibility test performed on the Sin-GG-HAP pellet confirmed the in-built bioactivity and biostability of the composites in simulated body fluid. Further, the Naringenin drug delivery profiles confirmed for the stable and controlled release of loaded drug from the Sin-GG-HAP composites (with 1, 5, and 10% of GG). We observed the highest release of 96% from the Sin-10% GG-HAP matrix in just 90 min and T50% of 416 h, and a mean diffusion time period of 459 h. Finally, the antimicrobial (using both gram-positive and gram-negative bacteria) and cytotoxic activities (with the use of healthy normal and diseased cells) of the drug-loaded and un-loaded GG-HAP composites confirm the potential role of the formed Sin-10% GG-HAP composite towards biomedical applications.

Increase in non-professional phagocytosis during the progression of cell cycle.

Homotypic or heterotypic internalization of another, either living or necrotic cell is currently in the center of research interest. The active invasion of a living cell called entosis and cannibalism of cells by rapidly proliferating cancers are prominent examples. Additionally, normal healthy tissue cells are capable of non-professional phagocytosis. This project studied the relationship between non-professional phagocytosis, individual proliferation and cell cycle progression. Three mesenchymal and two epithelial normal tissue cell lines were studied for homotypic non-professional phagocytosis. Homotypic dead cells were co-incubated with adherent growing living cell layers. Living cells were synchronized by mitotic shake-off as well as Aphidicolin-treatment and phagocytotic activity was analyzed by immunostaining. Cell cycle phases were evaluated by flow cytometry. Mesenchymal and epithelial normal tissue cells were capable of internalizing dead cells. Epithelial cells had much higher non-professional phagocytotic rates than mesenchymal cells. Cells throughout the entire cell cycle were able to phagocytose. The phagocytotic rate significantly increased with progressing cell cycle phases. Mitotic cells regularly phagocytosed dead cells, this was verified by Nocodazole and Colcemid treatment. Taken together, our findings indicate the ability of human tissue cells to phagocytose necrotic neighboring cells in confluent cell layers. The origin of the cell line influences the rate of cell-in-cell structure formation. The higher cell-in-cell structure rates during cell cycle progression might be influenced by cytoskeletal reorganization during this period or indicate an evolutionary anchorage of the process. Recycling of nutrients during cell growth might also be an explanation.

The BH3 mimetic (\xb1) gossypol induces ROS-independent apoptosis and mitochondrial dysfunction in human A375 melanoma cells in vitro

A major challenge in current cancer therapy is still the treatment of metastatic melanomas of the skin. BH3 mimetics represent a novel group of substances inducing apoptosis. In this study, we investigated the cytotoxic effect of (±) gossypol (GP), a natural compound from cotton seed, on A375 melanoma cells and the underlying biochemical mechanisms. To prevent undesired side effects due to toxicity on normal (healthy) cells, concentrations only toxic for tumor cells have been elaborated. Viability assays were performed to determine the cytotoxicity of GP in A375 melanoma and normal (healthy) cells. For the majority of experiments, a concentration of 2.5 µM GP was used resulting in a ROS-independent but caspase-dependent cell death of A375 melanoma cells. At this level, GP was non-toxic for normal human epidermal melanocytes. GP has a very short half-life, however, it was demonstrated that only the “parent” compound and not decomposition products are responsible for the cytotoxic effect in A375 melanoma cells. GP significantly decreased mitochondrial membrane potential accompanied by a Drp1-dependent loss of mitochondrial integrity (fragmentation) in tumor cells. Taken together, GP induced a ROS-independent intrinsic apoptosis leading to the conclusion that within a specific concentration range, GP may work as effective anticancer drug without harmful side effects.

Friend or Foe: Xenobiotic Activation of Nrf2 in Disease Control and Cardioprotection.

Nuclear factor erythroid 2-related factor 2 (Nrf2) is a transcription factor that governs a highly conserved pathway central to the protection of cells against various oxidative stresses. However, the biological impact of xenobiotic intervention of Nrf2 in physiological and pathophysiological conditions remains debatable. Activation of Nrf2 in cancer cells has been shown to elevate drug resistance and increase cell survival and proliferation, while inhibition of Nrf2 sensitizes cancer cells to drug treatment. On the other hand, activation of Nrf2 in normal healthy cells has been explored as a rather successful strategy for cancer chemoprevention. Selective activation of Nrf2 in off-target cells has recently been investigated as an approach for protecting off-target tissues from untoward drug toxicity. Specifically, induction of antioxidant response element genes via Nrf2 activation in cardiac cells is being explored as a means to limit the well-documented cardiotoxicity accompanied by cancer treatment with commonly prescribed anthracycline drugs. In addition to cancers, Nrf2 has been implicated in many other diseases including Alzheimer's and Parkinson's Diseases, diabetes, and cardiovascular disease. In this review, we discuss the roles of Nrf2 and its downstream target genes in the treatment of various diseases, and its recently explored potential for increasing the benefit: risk ratio of commonly utilized cancer treatments.

Cancer Vaccines: Antigen Selection Strategy.

Unlike traditional cancer therapies, cancer vaccines (CVs) harness a high specificity of the host’s immunity to kill tumor cells. CVs can train and bolster the patient’s immune system to recognize and eliminate malignant cells by enhancing immune cells’ identification of antigens expressed on cancer cells. Various features of antigens like immunogenicity and avidity influence the efficacy of CVs. Therefore, the choice and application of antigens play a critical role in establishing and developing CVs. Tumor-associated antigens (TAAs), a group of proteins expressed at elevated levels in tumor cells but lower levels in healthy normal cells, have been well-studied and developed in CVs. However, immunological tolerance, HLA restriction, and adverse events are major obstacles that threaten TAA-based CVs’ efficacy due to the “self-protein” characteristic of TAAs. As “abnormal proteins” that are completely absent from normal cells, tumor-specific antigens (TSAs) can trigger a robust immune response against tumor cells with high specificity and without going through central tolerance, contributing to cancer vaccine development feasibility. In this review, we focus on the unique features of TAAs and TSAs and their application in vaccines, summarizing their performance in preclinical and clinical trials.

Delivery of novel coumarin-dihydropyrimidinone conjugates through mixed polymeric nanoparticles to potentiate therapeutic efficacy against triple-negative breast cancer.

To date, most of the accessible therapeutic options are virtually non-responsive towards triple-negative breast cancer (TNBC) due to its highly aggressive and metastatic nature. Interestingly, chemotherapy reacts soundly in many TNBC cases compared to other types of breast cancer. However, the side effects of many chemotherapeutic agents are still under cross-examination, and thus prohibit their extensive uses. In this present study, we have developed a series of coumarin-dihydropyrimidinone conjugates (CDHPs) and subsequently their poly(lactic-co-glycolic acid) (PLGA)-PEG4000 mixed copolymer nanoparticles as excellent chemotherapeutic nanomedicine to control TNBC. Among all the synthesized CDHPs, CDHP-4 (prepared by the combination of EDCO with 3,4-difluorobenzaldehyde) showed excellent therapeutic effect on a wide variety of cancer cell lines, including TNBC. Besides, it can control the metastasis and stemness property of TNBC. Furthermore, the nano-encapsulation of CDHP-4 in a mixed polymer nanoparticle system (CDHP-4@PP-NPs) and simultaneous delivery showed much improved therapeutic efficacy at a much lower dose, and almost negligible side effects in normal healthy cells or organs. The effectiveness of the present therapeutic agent was observed both in intravenous and oral mode of administration in in vivo experiments. Moreover, on elucidating the molecular mechanism, we found that CDHP-4@PP-NPs could exhibit apoptotic, anti-migratory, as well as anti-stemness activity against TNBC cell lines through the downregulation of miR-138. We validated our findings in MDA-MB-231 xenograft chick embryos, as well as in 4T1-induced mammary tumor-bearing BALB/c mice models, and studied the bio-distribution of CDHP-4@PP-NPs on the basis of the photoluminescence property of nanoparticles. Our recent study, hence for the first time, unravels the synthesis of CDHP-4@PP-NPs and the molecular mechanism behind the anti-migration, anti-stemness and anti-tumor efficacy of the nanoparticles against the TNBC cells through the miR-138/p65/TUSC2 axis.

The disquisition of materialistic properties of tumor cells using a continuum model

The materialistic properties of a cell largely depend on the biological activities of the cell. These properties are the indicators of the maintenance of homeostasis in the human body. Any abnormal alterations in these properties can affect the day-to-day physiological functions of a healthy human body. In the present paper, we inquest the materialistic properties of a tumor cell in comparison with normal healthy somatic cells. Later continued research targets to mathematically model the tumor-immune system compartmentally by coupling with the materialistic property equation for the tumor cells. At the end of the paper, we present a continuum model for the spread of tumors as a function of the elastic modulus of the tumor cells. The model fabricated is a novel model that takes into account both the role of elastic modulus decrease as well as the role of immune cells in anti-tumor activities. The models available in the literature do not consider the role of the elastic property of the tumor cells into account.

Understanding Nanotechnology in the Treatment of Oral Cancer: A Comprehensive Review.

Oral cancer is the 11th most common cancer in the world with a high morbidity rate. Various conventional therapies have been used for the treatment of oral cancer such as surgery, radiotherapy, and chemotherapy used either alone or in combination but these have many limitations, making them unsuitable for treating oral cancer. Nanotechnology has been emerged out as an innovative tool in the field of oral cancer which has proved to provide effective results overcoming the limitations of conventional drug therapies. This system involves a nanoparticle drug delivery system based on a targeted therapy in which therapeutic drugs or agents act on the targeted cells without affecting normal healthy cells. Literature has shown that several nanoparticles, organic and inorganic nanoparticles, have been used as the drug delivery system in different types of oral cancers such as oral squamous cell carcinoma, cancer of the tongue, head, and neck cancers. Drugs like cisplatin, 5-fluorouracil, methotrexate, doxorubicin, etc., when coated with nano-polymers have shown better results compared with conventional drugs in oral cancer. Other nanoparticles such as liposomes, hydrogels, nanodiamonds, carbon rods, etc. have also been used with minimal side effects. This paper aims to review and discuss various nanotechnology systems in the field of oral cancer and to evaluate the efficacy of these systems in treating oral cancer compared with conventional drug delivery methods.

Protective role of herbal formulation-divine noni against cisplatin-induced cytotoxicity in healthy cells by activating Nrf2 expression: An in-vivo and in-vitro approach

BackgroundMorinda citrifolia (Noni) is an herbal remedy with multiple therapeutic properties such as antioxidant, anticancer, anti-inflammatory, antihypertensive effects due to rich in phytochemical constituents. PurposeTo evaluate chemoprotective effect of noni in dipping cisplatin (CP)-induced cytotoxicity in normal (healthy) cells through an antioxidant activity using in-vitro and in-vivo. MethodsChemoprotective study of fresh noni juice (NJ) and divine noni gold (DNG) at the doses (0.3125–10 mg/ml) were performed against CP (3.125–100 µM) induced cytotoxicity in normal cells (BEAS-2B) by Sulforhodamine B assay at 24 h and 48 h and the percentage of growth inhibition was calculated. Similarly, the protective effect of noni (360 mg/b.wt.) were investigated against CP (5 mg/ml) induced cardiotoxicity in healthy mice through the investigation of serum biochemical markers, endogenous antioxidant enzymes were interpreted. Finally, the chemoprotective mechanism of DNG was accomplished Nrf2 expression following CP challenged NIH-3T3 cells was performed by the western blot method. ResultsTreatment with noni at tested dose exhibited the least toxicity in normal cells (BEAS-2B). CP at the following concentrations induced cell death in time and dose-dependent manner. Whereas, noni revealed a significant cytoprotective effect against CP-induced cytotoxicity in the normal cells. Further in in-vivo study, treatment with noni at 360 mg/b.wt in CP injected animals attenuated cardiac injury as indicated by quenching the activities of serum biochemical markers, alleviated enzymatic and non-enzymatic antioxidant enzymes. Furthermore, noni attenuated CP induced toxicity by indirect antioxidant activity were proved by Nrf2 expression. ConclusionResults suggest that DNG is more potent than NJ. Findings confirmed that DNG protects against CP-induced toxicity by activating the Nrf2 expression. More studies are needed to comprehend the mechanisms of DNG cytoprotection.

Investigation of Anti-Proliferative Effects of Natural Products Quercetin Hydrate and Catechin Hydrate on Leukemia Lymphocytes

The major problem of conventional cancer therapy is lack of selectivity and induction of harmful side-effects on normal (healthy) cells and tissues. In the recent years, scientific efforts are focused to find a proper approach for highly selective influence on cell viability, as well as induction of cell death in cancer cells only. In this regard, natural herbal products are of great interest due to their low cytotoxicity to normal cells and tissues and their potential as supplements to conventional chemotherapeutics. It is well known that flavonoids exhibit various biological activities, such as anti oxidative, anti bacterial, anti inflammatory, anti viral and anti cancer, and may play a role in cancer prevention. In the present study, the effects of low concentrations of quercetin hydrate and catechin hydrate on cell viability of leukemia lymphocytes were investigated, in order to provide an experimental basis for their future incorporation into newly-synthesized biocompatible nano-formulations.

The selective anti-proliferative and pro-apoptotic effect of A. cherimola on MDA-MB-231 breast cancer cell line

BackgroundHerbal medicines have been a major target for numerous studies through the past years as an alternative treatment for cancer, mainly due to their minimal effects on normal healthy cells. Annona cherimola, popularly known as Cherimoya, is an edible natural fruit rich in phytochemical components and known to possess various biological activities. Previous studies have reported the anti-cancerous effect of A. cherimola ethanolic leaf extract (AELE) on leukemia. This study aims at studying the potential anti-cancer activity of this extract in vitro in two different breast cancer cell lines, namely MDA-MB-231 and MCF-7, in addition to investigating its toxicity on normal mesenchymal stem cells.MethodsThe anti-proliferative effect of AELE was evaluated via cell viability assay. Propidium iodide staining, Cell Death Detection ELISA and flow cytometry analysis of Annexin V binding were used to assess cell cycle progression, DNA fragmentation and apoptosis induction, respectively. Protein expression was determined via Western Blot analysis to decipher the underlying apoptotic molecular mechanism induced upon AELE treatment.ResultsThe anti-proliferative effect of the extract was found to be selective on the triple-negative breast cancer cell line (MDA-MB-231) in a time- and dose-dependent manner with an IC50 of 390.2 μg/mL at 48 h, with no cytotoxic effects on normal murine mesenchymal stem cells. The pro-apoptotic effect was confirmed by the increase in cellular and DNA fragmentation, flipping of the phosphatidylserine moiety to the outer leaflet, and the increase in Annexin V binding. The underlying molecular mechanism revealed the involvement of the mitochondrial pathway, as shown by alterations in mitochondrial permeability and the upregulation of cytochrome c expression.ConclusionAll the data presented in our study suggest that AELE exhibits a selective anti-proliferative and pro-apoptotic effect on the chemo-resistant MDA-MB-231 breast cancer cells, providing evidence for the anti-tumor effects of A. cherimola.

Surface receptor-mediated targeted drug delivery systems for enhanced cancer treatment: A state-of-the-art review.

Enhanced cancer treatment remains as one of the focused areas for researchers around the world. Hence, the progress in this direction will be a challenge and an opportunity in, inter-disciplinary field to mitigate the suffering of millions in the upcoming decades. As we see, cancer death rate has also progressively increased despite the current impressive treatment regimens but also due to the non-availability of vaccines and the re-occurring of cancer in substantially recovered patients. Currently, numerous treatment strategies like surgical removal of solid tumors followed by radiation with a combination of immunotherapy/chemotherapy by the researchers and clinicians are routinely being followed. However, recurrence and distant metastasis often occur following radiation therapy, commonly due to the generation of radio-resistance through deregulation of the cell cycle, cell death, and inhibition of DNA damage repair mechanisms. Thus, chemotherapeutic/immunotherapeutic treatment systems have progressed remarkably in the latest years owing to destroying tumors, noninvasive, and affordable charge of therapy. But, traditional chemotherapeutic approaches target the DNA of mutated and normal healthy cells, resulting in a significantly increased risk of toxicity and drug resistance. Thus, many receptors targeted therapies are in the developmental phase of discovery. Cancer cells have a specialized set of surface receptors that provide potential targets for cancer therapeutics. Cell surface receptor-dependent endocytosis is well a known major mechanism for the internalization of macromolecular drugs. This review emphasizes the recent development of several surface receptors mediated cancer-targeting approaches for the effective delivery of various therapeutic formulations.

Multiple Emulsions as a Biomaterial-based Delivery System for the Controlled Release of an Anti-cancer Drug

This paper focuses on developing multiple emulsions as a pH-responsive drug delivery system, for glioblastoma multiforme-GBM therapy, with reduced toxicity to healthy cells. The multiple emulsions with a stimuli-responsive biopolymer (CMC- sodium carboxymethylcellulose) were prepared in a Couette-Taylor flow contactor. As an external stimulus, the difference in pH of the cancer environment, and normal tissue, was investigated by adding salts as a triggering agent. The cancer cell lines of glioblastoma multiforme were investigated: U87MG, LN229, T98G, in order to verify emulsions’ components cytotoxicity to cells. Also normal (healthy) cells, K21-fribroblast, were analysed. Rhodamine B was used as a model drug instead of the clinically used chemotherapeutics (e.g. doxorubicin) in oncology. Results showed that multiple emulsions by themselves had no adverse effect on the viability of investigated cells, excluding one cell line: LN229. The control and modulated release rates of a model drug, by stimuli-responsive biopolymer, were established. Results confirmed the possibility of controlling the release rates of a drug in the acidic environment of the cancer cells. The proposed multiple emulsion could be explored for the potential delivery of chemotherapeutics in GBM therapy.

The dual protease inhibitor lopinavir/ritonavir (LPV/r) exerts genotoxic stress on lung cells

The Sub-Saharan countries, particularly South Africa has the largest number of people living with HIV, accompanied by the largest antiretroviral treatment (ART) programme in the world. The Highly Active Antiretroviral Treatment (HAART) is the most effective regimen against HIV/AIDS and has improved the lifespan and quality of life of HIV positive patients. HAART has also led to a decrease in the incidence of AIDS defining cancers (ADCs) while there is an increased incidence of the non-AIDS Defining Cancers (NADCs), such as lung cancer in the HAART era. The association between lung tumourigenesis and the use of HAART components such as the dual protease inhibitor (PI) lopinavir/ritonavir (LPV/r) is poorly understood. Using cell and molecular biological approaches, this study aimed at elucidating the effects of LPV/r on the regulation of the cell cycle related genes in normal (MRC-5) and adenocarcinoma (A549) lung cells. Initially, the nuclear integrity of these cells in response to LPV/r was determined using DAPI staining. The effect of LPV/r on cell cycle genes was evaluated through the use of a RT2 PCR gene array of 84 genes related to the cell cycle signaling pathway. The PCR array data was validated by Real-Time Quantification PCR (RT-qPCR). Ingenuity Pathway Analysis (IPA) bio-informatics tool was employed to disclose the molecular mechanism/s observed at cellular and gene expression levels. Loss of nuclear integrity and the upregulation of the p53 DNA damage response (DDR) pathway was revealed by DAPI staining, differential gene expression and IPA core analysis. Furthermore, MAD2L2 and AURKB which also play a role in the DDR pathway were shown to be differentially expressed. The activation of the CASP3 gene in response to LPV/r in A549 cells was also observed. The findings of this study suggest genotoxic properties of LPV/r in healthy normal lung fibroblasts cells and anti-tumour properties in the A549 cells.

Study of potent cytotoxic activity of Helleborus cyclophyllus Boiss against a human adenocarcinoma cell line.

Helleborus cyclophyllus Boiss is a rhizomatous plant species, with strong allelochemical properties, that has been used since ancient times for its therapeutic properties. In the present study we investigated the ability of an aqueous-soluble fraction of the methanol extract of H. cyclophyllus Boiss leaves, to induce apoptotic cell death on A549 human bronchial epithelial adenocarcinoma cells. A primary human lung fibroblasts' cell line was used as a model of normal-healthy cells for comparison. Cell morphology was examined after appropriate staining, cytotoxic activity of the extract was determined by the MTT assay, the type of cell death was analyzed by flow cytometry, confirmation of apoptosis was evaluated with the analysis of caspase-3, PARP1 by western blotting, while the chemical composition was assessed by liquid chromatography-tandem mass spectrometry (LC-MS/MS). H. cyclophyllus Boiss extract was selectively active on A549 cells inducing significant morphological changes, even at low concentrations. Characteristic morphological alterations included the release of vesicular formations from A549 cell membranes (ectosomes), detachment of cells from their substrate, generation of a large vesicle into the cytoplasm (thanatosome) and the formation of apoptotic bodies. The selective apoptotic action on treated cells was also confirmed by biochemical criteria. Low concentrations, however, did not affect normal cells. The phytochemical analysis of the extract revealed the presence of cardiac glucosides, bufadienolides and phytoecdysteroids. To the best of our knowledge, the above-mentioned sequences of events leading selectively cancer cells to apoptosis, has not been reported before.