Size-Based Differentiation of Cancer and Normal Cells by a Particle Size Analyzer Assisted by a Cell-Recognition PC Software.

Actin-based biomechanical features of suspended normal and cancer cells

Temporins are small antimicrobial peptides which have cytotoxic effects against different cancerous cell lines. However, their cytotoxicity against normal cells has been reported in some studies. Designing new temporin analogs with selective cytotoxicity against cancerous cell lines is considered a goal in drug development. In this regard, four new temporin-1CEa analogs (either C-terminally α-amidated or not) with increased net positive charge, as well as the potential of hydrogen bond formation were designed and their selective cytotoxicity against prostate carcinoma cell line (LNCaP) was evaluated. MTT assay was employed to test the simultaneous effects of the modifications on the cytotoxicity of the analogs against both normal (HFFF2) and cancerous (LNCaP) cell lines. Then, the in vitro selectivity index of each analog was determined using the equation of selectivity index (SI= IC50 of peptide against normal cell line / IC50 of peptide against cancerous cell line). The structural modifications of temporin-1CEa (increasing the net positive charge and removing C-terminally amidated group, as well as the potential of hydrogen bond formation on the non-polar face of α-helical structure) reduced the cytotoxicity of K2K3W15-COOH analog against both cancerous (LNCaP) and normal (HFFF2) cell lines. However, the modifications resulted in improving the in vitro selectivity index of K2K3W15-COOH compared to the temporin-1CEa (1.67 vs. 1.33, respectively). The results of the present study indicated that some structural modifications may lead to a diminution in the cytotoxicity of new analogs against cancerous cells. However, the same modifications can be associated with better selectivity indexes of new analogs due to a further decrease in their cytotoxicity against normal cells.

BackgroundCancer is an international health problem, and the search for effective treatments is still in progress. Peptide therapy is focused on the development of short peptides with strong tumoricidal activity and low toxicity. In this study, we investigated the efficacy of a myxoma virus peptide analogue (RRM-MV) as a candidate for skin cancer therapy. RRM-MV was designed using the Resonant Recognition Model (RRM) and its effect was examined on human skin cancer and normal human skin cells in vitro.MethodsCell cultures were treated with various concentrations of the peptides at different incubation intervals. Cellular morphological changes (apoptosis and necrosis) were evaluated using confocal laser scanning microscopy. The cytotoxic effects of RRM-MV on human skin cancer and normal human skin cells were quantitatively determined by cytotoxicity and cell viability assays. The effect on human erythrocytes was also determined using quantitative hemolysis assay. DNA fragmentation assay was performed to detect early apoptotic events in treated cancer cells. Furthermore, to investigate the possible cell signalling pathway targeted by the peptides treatment, the levels of p-Akt expression in skin cancer and normal cells were detected by immunoblotting.ResultsOur results indicate that RRM-MV has a dose-dependent toxic effect on cancer cells only up to 18 h. The immunoblotting results indicated that the RRM-MV slightly increased p-Akt expression in melanoma and carcinoma cells, but did not seem to affect p-Akt expression in normal skin cells.ConclusionsRRM-MV targets and lethally harms cancer cells and leaves normal cells unharmed. It is able to reduce the cancer cell viability, disrupting the LDH activity in cancer cells and can significantly affect cancer progression. Further investigation into other cell signalling pathways is needed in the process leading to the in vivo testing of this peptide to prove its safety as a possible effective treatment for skin cancer.

Effects of continuous wave and fractionated diode laser on human fibroblast cancer and dermal normal cells by zinc phthalocyanine in photodynamic therapy: A comparative study

The antimicrobial, anti-inflammatory and tissue-stimulating effects of cold argon atmospheric plasma (CAAP) accelerate its use in various fields of medicine. Here, we investigated the effects of CAAP at different radiation doses on mesenchymal stem cells (MSCs) and human osteosarcoma (MNNG/HOS) cells. We observed an increase in the growth rate of MSCs at sufficiently low irradiation doses (10–15 min) of CAAP, while the growth of MNNG/HOS cells was slowed down to 41% at the same irradiation doses. Using flow cytometry, we found that these effects are associated with cell cycle arrest and extended death of cancer cells by necrosis. Reactive oxygen species (ROS) formation was detected in both types of cells after 15 min of CAAP treatment. Evaluation of the genes’ transcriptional activity showed that exposure to low doses of CAAP activates the expression of genes responsible for proliferation, DNA replication, and transition between phases of the cell cycle in MSCs. There was a decrease in the transcriptional activity of most of the studied genes in MNNG/HOS osteosarcoma cancer cells. However, increased transcription of osteogenic differentiation genes was observed in normal and cancer cells. The selective effects of low and high doses of CAAP treatment on cancer and normal cells that we found can be considered in terms of hormesis. The low dose of cold argon plasma irradiation stimulated the vital processes in stem cells due to the slight generation of reactive oxygen species. In cancer cells, the same doses evidently lead to the formation of oxidative stress, which was accompanied by a proliferation inhibition and cell death. The differences in the cancer and normal cells’ responses are probably due to different sensitivity to exogenous oxidative stress. Such a selective effect of CAAP action can be used in the combined therapy of oncological diseases such as skin neoplasms, or for the removal of remaining cancer cells after surgical removal of a tumor.

e22113 Background: It is estimated that low dose radiation can increase the risk of cancer as well as mutations. However, the interaction of low dose radiation with caffeine has not been adequately investigated. We investigated the effects of caffeine on low dose- gamma-radiation-induced chromosomal damage in human T leukemia cells (Jurket T-cells) and two normal human fibroblast cell lines (AG1522 and GM 2149). Method: Jurkat cells were maintained in RPMI 1640 medium and fibroblast in alpha-minimal essential medium (MEM) All cells were incubated at 37o C in a humidified atmosphere of 5% CO2 in air. Cells from the exponential phase were treated with 1 mg/ml caffeine ( control cells received same amount of solvent) and irradiated with low doses (3, 5, 10, 20 and 40 cGy,), using a 137 Cs-gamma radiation source. Colcemid at a concentration of 0.1 μg/ml was added to every flask. Cells were fixed in methanol: acetic acid solution and stained with Giemsa. 100 irradiated and un-irradiated metaphase- like cells were scored for chromatid-type aberrations. Results: Low dose gamma-radiation increased the levels of chromatid breaks(dose dependent) in both normal and cancer cells; however, cancer cells appeared to be more sensitive than the normal cells. Caffeine treatment markedly increased chromatid aberrations in Jurkat T-cells at all radiation doses but not in normal cells. Previously, we reported that caffeine eliminates gamma-ray-induced G2 delay in other human tumor cells but not normal cells (Jha, et.al., Radiat. Res. 157, 26–31, 2002). Conclusions: The mechanisms that may underlie this differential effect of caffeine in cancer and normal cells are unknown, but if one result of a G2 delay is to allow more time for chromosome breakage rejoining processes to occur, then elimination of this delay by caffeine in tumor cells but not normal cells might account for the difference. To the extent these observations are generally true for tumor vs normal cells, the differential sensitization could have an impact in improving the efficacy of radiation therapy. No significant financial relationships to disclose.

Cellular Traction Forces and Locations of Adhesion Site Regulate Cell Functions

We report the detection of oral cancer cells using Raman spectroscopy as a noninvasive diagnostic tool. Gold nanoparticles, synthesized by using sodium citrate as a reducing agent, were used as surface‐enhanced Raman scattering substrates to detect human oral squamous cell carcinoma cell lines (SAS and SCC4) with fibroblasts cell lines as the control. We have observed the characteristic Raman signal of adenine at 735 cm−1, which indicates the oral cancer from nucleic acid. The result showed that there were significant differences (p < .05) between oral cancer cell and normal fibroblast cell lines. To prove that adenine is a robust biomarker for cancer cells, we have also compared the Raman spectra of esophageal cancer (CE48T and CE81T) and normal (Het‐1A) cells. Significant differences (p < .05) were also observed between esophageal cancer and normal cells. The detection of adenine Raman signal from cell lines has been proven to be potential to establish a highly distinguishable Raman diagnostic method for cancer detection.

In an attempt to improve the chemotherapy of acute leukemia in adults, the author has studied the induction therapy on 63 cases with acute leukemia with Neocarzinostatin (NCS; N) in combination with conventional antileukemic agents; Vincristine (V), Daunorubicin (D), Cytosine Arabinoside (A), 6-Mercaptopurine-Riboside (M) and Prednisolone (P). Eighteen cases with acute lymphocytic leukemia (ALL) was treated with protocol A (NVMP, NDMP, NAMP). Out of 45 cases with acute non-lymphocytic leukemia (ANLL), 25 was treated with protocol A and 20 treated with protocol B (NADP, NAVP), and obtained the following results. 1) Of all 63 cases 51 (81.6%) attained complete remission and 9 (14.3%) partial remission. 2) Complete remission was obtained in 15 cases (83.3%) out of 18 with ALL and 36 cases (80.0%) of 45 with ANLL; the latter cases with ANLL consisted of 18 of 25 cases (72.0%) treated with protocol A and 18 of 20 cases (90.0%) with protocol B. 3) Refractory varieties included cases with acute myelogenous leukemia who had passed through atypical hematological findings, cases with monocytic leukemia, cases with WBC count over 50,000 per μl or with nucleated cell count in bone marrow over 500,000 per μl before treatment and cases over 50 years old. 4) In this induction therapy with NCS in combination, antileukemic effects was remarkable, while suppression on normal hematopoiesis, especially on thrombocytes, was very slight. 5) As to the other side effects no noteworthy disturbances other than occasional nausea, anorexia and liver damage in protocol B were observed. From these results it can be concluded that NCS in combination is one of the most potent remission induction therapies in acute leukemia in adults.

Detection of Her2-overexpressing cancer cells using keyhole shaped chamber array employing a magnetic droplet-handling system

Millions of deaths occur each year due to the late diagnosis of abnormal cellular growth within the body. However, the devastating impact of this can be significantly reduced if cancer metastasis is detected early through the use of enzymatic biomarkers. Among several biomarkers, γ-glutamyltranspeptidase (GGT) stands out as a member of the aminopeptidase family. It is primarily found on the surface of cancer cells such as glioma, ovarian, lung, and prostate cancer, without being overexpressed in normal cells or tissues. Recent years have witnessed significant progress in the field of cancer monitoring and imaging. Fluorescence sensing techniques have been employed, utilizing organic small molecular probes with enzyme-specific recognition sites. These probes emit a fluorescent signal upon interacting with GGT, enabling the imaging, identification, and differentiation of normal and cancerous cells, tissues, and organs. This review article presents a concise overview of recent progress in fluorescent probes developed for the selective detection of GGT, focusing on their applications in cancer imaging. It highlights the observed alterations in the fluorescence and absorption spectra of the probes before and after interaction with GGT. Additionally, the study investigates the changes in the probe molecule's structure following enzyme treatment, evaluates the sensor's detection limit, and consolidated imaging studies conducted using confocal fluorescence analysis. This comprehensive survey is expected to contribute to the advancement of sensing techniques for biomarker detection and cancer imaging, providing valuable insights for refining methodologies and inspiring future developments in this field.

A major challenge in cancer therapy is to develop drugs that kill tumor cells without being toxic to normal cells. More than 15 years ago, sulindac and related compounds were shown to have anti-tumor activity that did not appear to be related to sulindac's anti-inflammatory activity. This proposal is based on the unique properties of sulindac that we have observed that are unrelated to its NSAID activity. This drug can protect certain normal cells, such as lung, retinal pigment epithelial cells (RPE) and cardiac cells, against oxidative stress, but markedly enhance the killing of all cancer cells tested in the presence of oxidative stress. The protection of cardiac cells against ischemia/reperfusion oxidative damage by sulindac has been shown to be due to an ischemic preconditioning mechanism. We believe that sulindac, in the presence of any compound that affects mitochondrial respiration resulting in ROS formation, causes cancer cell death due to apoptosis. Our hypothesis is that sulindac, and other ischemic preconditioning agent (IPC) in combination with agents that cause oxidative stress, initiate a protective preconditioning response to oxidative stress in normal cells, but trigger an apoptotic response to oxidative stress in cancer cells. Our preliminary results have shown a most important new and unique observation, that other known ischemic preconditioning agents, such as sildenafil can substitute for sulindac and enhance the killing of cancer cells exposed to oxidative stress. We tested for the most active drug combination of IPC agents on the viability of three different cancer cell lines, retinoblastoma (Rb), lung (A549) and glioblastoma (U87) using either sulindac or sildenafil in combination with anti-cancer agents that cause mitochondrial dysfunction such as doxorubicin (DOX), DCA and arsenic trioxide (As2O3). Our results show that enhanced cancer killing was seen with all the three cancer cell lines using either sulindac/sildenafil with both DOX and DCA whereas enhanced killing with As2O3 was observed only in A549 cells highlighting the metabolic differences in the cell lines. In order to confirm that sulindac and sildenafil are acting as preconditioning agents, we used normal RPE cells and exposed them to UV radiation to produce oxidative damage in the presence or absence of sulindac/sildenafil. Both sulindac and sildenafil significantly protected the RPE cells against UV induced damage. We obtained preliminary evidence on the role of PKC (Protein Kinase C) isoforms in the differential response of normal and cancer cells to IPC agents. Our results indicate that PKC delta is involved in the enhanced cancer killing, but PKC epsilon is required for the protection of normal cells against oxidative stress. In summary, our studies indicate for the first time that IPC agents make cancer cells more sensitive to oxidative stress and that the combination of an IPC agent with an oxidizing agent, or compounds that will perturb mitochondrial respiration, could have important therapeutic value. Our novel concept of the effect of IPC agents on cancer cells possess a huge potential to develop individualized therapeutic combinations for different cancers based on the metabolic differences in each cancer cell line. Citation Format: Shailaja Kesaraju, Arunodoy Sur, Kasirajan Ayyanathan, Herbert Weissbach. Ischemic preconditioning agents sensitize cancer cells to oxidative stress. [abstract]. In: Proceedings of the Eleventh Annual AACR International Conference on Frontiers in Cancer Prevention Research; 2012 Oct 16-19; Anaheim, CA. Philadelphia (PA): AACR; Cancer Prev Res 2012;5(11 Suppl):Abstract nr A61.

Photodynamic diagnosis and therapy (PDD & PDT) of cancer use generally photosensitizers for the discrimination of normal and cancer cells. In contrast we have studied the autofluorescence spectra of normal and cancer cells of rat using a tunable optical parametric oscillator (OPO) laser system as a light excitation source. WKA rat fibroblast (normal cell) and H-ras oncogene-transfected rat fibroblast (cancer cell) were used in this study. The samples were mounted in a quartz cuvette. The absorption peaks were observed at about 205 nm for both normal and cancer cells. On the other hand, the specific emission peaks were observed at 450 nm and 455 nm for normal and cancer cells, respectively, whereas the autofluorescence intensity of cancer cells was 2.5 times larger than that of normal cells. These changes are attributed to changes in proteins in the cancer cell and to the increase of nuclei. This suggests the possibility that the autofluorescence of cancer cell can be used to diagnose a human malignant tumor, so that it is concluded that normal and cancer cells can be discriminated by means of autofluorescence analysis.

<div>Abstract<p>Cancer cells typically experience higher oxidative stress than normal cells, such that elevating pro-oxidant levels can trigger cancer cell death. Although pre-exposure to mild oxidative agents will sensitize cancer cells to radiation, this pre-exposure may also activate the adaptive stress defense system in normal cells. Ascorbic acid is a prototype redox modulator that when infused intravenously appears to kill cancers without injury to normal tissues; however, the mechanisms involved remain elusive. In this study, we show how ascorbic acid kills cancer cells and sensitizes prostate cancer to radiation therapy while also conferring protection upon normal prostate epithelial cells against radiation-induced injury. We found that the NF-κB transcription factor RelB is a pivotal determinant in the differential radiosensitization effects of ascorbic acid in prostate cancer cells and normal prostate epithelial cells. Mechanistically, high reactive oxygen species concentrations suppress RelB in cancer cells. RelB suppression decreases expression of the sirtuin SIRT3 and the powerful antioxidant MnSOD, which in turn increases oxidative and metabolic stresses in prostate cancer cells. In contrast, ascorbic acid enhances RelB expression in normal cells, improving antioxidant and metabolic defenses against radiation injury. In addition to showing how RelB mediates the differential effects of ascorbic acid on cancer and normal tissue radiosensitivities, our work also provides a proof of concept for the existence of redox modulators that can improve the efficacy of radiotherapy while protecting against normal tissue injury in cancer settings. <i>Cancer Res; 77(6); 1345–56. ©2017 AACR</i>.</p></div>

Current cancer diagnostic methods lack the ability to quickly, simply, efficiently, and inexpensively screen cancer cells from a mixed population of cancer and normal cells. Methods based on biomarkers are unreliable due to complexity of cancer cells, plasticity of markers, and lack of common tumorigenic markers. Diagnostics are time intensive, require multiple tests, and provide limited information. In this study, we developed a novel wicking fiber device that separates cancer and normal cell types. To the best of our knowledge, no previous work has used vertical wicking of cells through fibers to identify and isolate cancer cells. The device separated mouse mammary tumor cells from a cellular mixture containing normal mouse mammary cells. Further investigation showed the device separated and isolated human cancer cells from a heterogeneous mixture of normal and cancerous human cells. We report a simple, inexpensive, and rapid technique that has potential to identify and isolate cancer cells from large volumes of liquid samples that can be translated to on-site clinic diagnosis.