scholarly journals Analysis of Single Nucleotide-Mutated Single-Cancer Cells Using the Combined Technologies of Single-Cell Microarray Chips and Peptide Nucleic Acid-DNA Probes

Micromachines ◽  
2020 ◽  
Vol 11 (7) ◽  
pp. 628
Author(s):  
Hajime Shigeto ◽  
Eriko Yamada ◽  
Mizuki Kitamatsu ◽  
Takashi Ohtsuki ◽  
Akira Iizuka ◽  
...  

Research into cancer cells that harbor gene mutations relating to anticancer drug-resistance at the single-cell level has focused on the diagnosis of, or treatment for, cancer. Several methods have been reported for detecting gene-mutated cells within a large number of non-mutated cells; however, target single nucleotide-mutated cells within a large number of cell samples, such as cancer tissue, are still difficult to analyze. In this study, a new system is developed to detect and isolate single-cancer cells expressing the T790M-mutated epidermal growth factor receptor (EGFR) mRNA from multiple non-mutated cancer cells by combining single-cell microarray chips and peptide nucleic acid (PNA)-DNA probes. The single-cell microarray chip is made of polystyrene with 62,410 microchambers (31-40 µm diameter). The T790M-mutated lung cancer cell line, NCI-H1975, and non-mutated lung cancer cell line, A549, were successfully separated into single cells in each microchambers on the chip. Only NCI-H1975 cell was stained on the chip with a fluorescein isothiocyanate (FITC)-conjugated PNA probe for specifically detecting T790M mutation. Of the NCI-H1975 cells that spiked into A549 cells, 0–20% were quantitatively analyzed within 1 h, depending on the spike concentration. Therefore, our system could be useful in analyzing cancer tissue that contains a few anticancer drug-resistant cells.

Micromachines ◽  
2021 ◽  
Vol 12 (10) ◽  
pp. 1147
Author(s):  
Yugyung Jung ◽  
Minkook Son ◽  
Yu Ri Nam ◽  
Jongchan Choi ◽  
James R. Heath ◽  
...  

Cancer is a dynamic disease involving constant changes. With these changes, cancer cells become heterogeneous, resulting in varying sensitivity to chemotherapy. The heterogeneity of cancer cells plays a key role in chemotherapy resistance and cancer recurrence. Therefore, for effective treatment, cancer cells need to be analyzed at the single-cell level by monitoring various proteins and investigating their heterogeneity. We propose a microfluidic chip for a single-cell proteomics assay that is capable of analyzing complex cellular signaling systems to reveal the heterogeneity of cancer cells. The single-cell assay chip comprises (i) microchambers (n = 1376) for manipulating single cancer cells, (ii) micropumps for rapid single-cell lysis, and (iii) barcode immunosensors for detecting nine different secretory and intracellular proteins to reveal the correlation among cancer-related proteins. Using this chip, the single-cell proteomics of a lung cancer cell line, which may be easily masked in bulk analysis, were evaluated. By comparing changes in the level of protein secretion and heterogeneity in response to combinations of four anti-cancer drugs, this study suggests a new method for selecting the best combination of anti-cancer drugs. Subsequent preclinical and clinical trials should enable this platform to become applicable for patient-customized therapies.


2020 ◽  
Vol 10 (3) ◽  
pp. 109 ◽  
Author(s):  
Hee Sung Chae ◽  
Minchan Gil ◽  
Subbroto Kumar Saha ◽  
Hee Jeung Kwak ◽  
Hwan-Woo Park ◽  
...  

Lung cancer remains the most dangerous type of cancer despite recent progress in therapeutic modalities. Development of prognostic markers and therapeutic targets is necessary to enhance lung cancer patient survival. Sestrin family genes (Sestrin1, Sestrin2, and Sestrin3) are involved in protecting cells from stress. In particular, Sestrin2, which mainly protects cells from oxidative stress and acts as a leucine sensor protein in mammalian target of rapamycin (mTOR) signaling, is thought to affect various cancers in different ways. To investigate the role of Sestrin2 expression in lung cancer cells, we knocked down Sestrin2 in A549, a non-small cell lung cancer cell line; this resulted in reduced cell proliferation, migration, sphere formation, and drug resistance, suggesting that Sestrin2 is closely related to lung cancer progression. We analyzed Sestrin2 expression in human tissue using various bioinformatic databases and confirmed higher expression of Sestrin2 in lung cancer cells than in normal lung cells using Oncomine and the Human Protein Atlas. Moreover, analyses using Prognoscan and KMplotter showed that Sestrin2 expression is negatively correlated with the survival of lung cancer patients in multiple datasets. Co-expressed gene analysis revealed Sestrin2-regulated genes and possible associated pathways. Overall, these data suggest that Sestrin2 expression has prognostic value and that it is a possible therapeutic target in lung cancer.


2014 ◽  
Vol 2014 ◽  
pp. 1-8 ◽  
Author(s):  
Lérida Liss Flores Villavicencio ◽  
Gustavo Cruz-Jiménez ◽  
Gloria Barbosa-Sabanero ◽  
Carlos Kornhauser-Araujo ◽  
M. Eugenia Mendoza-Garrido ◽  
...  

The effects of organic and inorganic forms of selenium (Se) on human cells have been extensively studied for nutritional concentrations; however, to date, little is known about the potential toxicity at supranutritional levels. In the present study we determined the effects of sodium selenite (SSe) and selenomethionine (SeMet) on cell growth and intracellular structures in lung cancer cells exposed at Se concentrations between 0 and 3 mM. Our results showed that SSe affected cell growth more rapidly than SeMet (24 h and 48 h, resp.). After 24 h of cells exposure to 0.5, 1.5, and 3 mM SSe, cell growth was reduced by 10, 50, and 60%, as compared to controls. After 48 h, nuclear fragmentation was evident in cells exposed to SSe, suggesting an induction to cell death. In contrast, SeMet did not affect cell proliferation, and the cells were phenotypically similar to controls. Microtubules and microfilaments structures were also affected by both Se compounds, again SSe being more toxic than SeMet. To our knowledge, this is the first report on the differential effects of organic and inorganic Se in supranutritional levels in lung cancer cells.


Sensors ◽  
2017 ◽  
Vol 17 (10) ◽  
pp. 2410 ◽  
Author(s):  
Shohei Yamamura ◽  
Eriko Yamada ◽  
Fukiko Kimura ◽  
Kumiko Miyajima ◽  
Hajime Shigeto

2011 ◽  
Vol 66 (9-10) ◽  
pp. 477-484 ◽  
Author(s):  
Wenfeng Huang ◽  
Kun Zou ◽  
Bin Xiong

Trillium tschonoskii Maxim. has been used to treat several diseases including cancers in folk medicine. However, the mechanisms responsible for T. tschonoskii extract-induced apoptosis are not clear. This study was mainly undertaken to identify the major biochemical changes in a lung cancer cell line upon treatment with an T. tschonoskii extract (TTME), and to investigate the functional relationship between these changes. The n-butanol extract was used to evaluate the mechanism of induction of apoptosis in A549 human lung cancer cells and its effects on mitochondrial function and production of reactive oxygen species (ROS). The n-butanol extract of T. tschonoskii has cytotoxic, antiproliferative, and morphological effects on the lung cancer cell line. T. tschonoskii mainly leads to apoptosis of cancer cells with a concomitant increase in the release of cytochrome c and a loss of mitochondrial membrane potential in a dose-dependent manner. A rapid increase in the level of intracellular ROS and an accumulation of cells in the G2/M and S phase of the cell cycle were also observed in treated cells. These observations suggest that the n-butanol extract of T. tschonoskii has promising anticancer activities, which could be useful in cancer treatment.


2019 ◽  
Author(s):  
Ting Wang ◽  
Yiqian Liang ◽  
Asmitananda Thakur

Abstract Background S100 calcium binding protein A2 (S100A2) has been confirmed to have an abnormal expression in lung cancer and is associated with a better disease-free internal of lung cancer patients. Our previous studies on S100A2 in lung cancer concentrated on the clinical roles of this protein in lung cancer, finding that S100A2 increasingly expressed in the sera, tissues and plural effusion of lung cancer patients. This study emphasizes its value in the lung cancer cell line.Methods We constructed a S100A2 expression lentivirus vector, then transfected it and blank vector into the Calu-6 lung cancer cell line respectively. After the successful transfection, (which was confirmed by RT-PCR and Western-blot), we used MTT, transwell and flow cytometric analysis to compare the differences in cell proliferation, cell migration, cell invasion, cell apoptosis and cell cycle among the three groups (Calu-6, Calu/neo, Calu-6/S100A2).Results Calu-6 lung cancer cells showed a shift from G1 to S phase after being transfected with S100A2, compared with the control groups. Additionally, Calu-6/S1000A2 cells had enhanced abilities of invasion and down-abilities of apoptosis in contrast with the blank groups (P<0.05). However, there were no significant difference among these three group in the cell behaviors of migration and proliferation (P>0.05).Conclusion Our results firstly indicate that S100A2 has a positive influence on the biological characteristics of Calu-6 lung cancer cell line, including cell division, invasion and apoptosis inhibition. It may play a significant role in the genesis and progression of lung cancer.


Author(s):  
Andreas Ulrich Lindner ◽  
Manuela Salvucci ◽  
Elizabeth McDonough ◽  
Sanghee Cho ◽  
Xanthi Stachtea ◽  
...  

AbstractCancer cells’ ability to inhibit apoptosis is key to malignant transformation and limits response to therapy. Here, we performed multiplexed immunofluorescence analysis on tissue microarrays with 373 cores from 168 patients, segmentation of 2.4 million individual cells, and quantification of 18 cell lineage and apoptosis proteins. We identified an enrichment for BCL2 in immune, and BAK, SMAC, and XIAP in cancer cells. Ordinary differential equation-based modeling of apoptosis sensitivity at single-cell resolution was conducted and an atlas of inter- and intra-tumor heterogeneity in apoptosis susceptibility generated. Systems modeling at single-cell resolution identified an enhanced sensitivity of cancer cells to mitochondrial permeabilization and executioner caspase activation compared to immune and stromal cells, but showed significant inter- and intra-tumor heterogeneity.


Nanomedicine ◽  
2019 ◽  
Vol 14 (17) ◽  
pp. 2355-2371 ◽  
Author(s):  
Shivani Thoidingjam ◽  
Ashu Bhan Tiku

Aim: Present work was undertaken to fabricate iron oxide nanoparticles (IONPs) using a green approach for increased therapeutic efficacy. Materials & methods: Two types of IONPs were synthesized, one without any coating (IONPUC) and other coated with Phyllanthus emblica (Amla) fruit extract (IONPA). Both the IONPs were characterized using different techniques and therapeutic efficacy was evaluated in A549 human lung cancer cell line. Results: IONPA were smaller in size with better dispersibility compared with IONPUC. They induced increased reactive oxygen species production, higher DNA damage and apoptosis, which resulted in increased toxicity to cancer cells in comparison to IONPUC. Conclusion: Higher uptake of IONPA and active components coating the surface, may be responsible for the increased therapeutic efficacy in cancer cells.


2021 ◽  
Vol 15 (5) ◽  
pp. 1282-1284
Author(s):  
Moein Shaneh

Chemotherapy is a type of cancer treatment in which the lack of selective cytotoxicity often leads to intolerable side effects. Today, the use of medicinal plants is essential in treating cancer due to their fewer side effects. Lagenaria siceraria Standl is critical for cytotoxicity studies due to its polyphenolic, cucurbitacins, pectin, flavonoids, and saponin compounds. In this study, the cytotoxic effects of plant fruit extract were investigated on lung cancer cell lines. To this end, the hydroalcoholic extract of the plant fruit was initially prepared by the percolation method. Then, the effects of solutions containing samples with different concentrations (5000, 500, 1000, 100, 100, 250, 10, 1, 0.1μg.ml-1) were investigated by MTT assay on lung cancer cell line (A549). Cisplatin was considered as a positive control. Statistical calculations were carried out using Prism V.3 software to compare IC50, and the data were analyzed by analysis of variance (ANOVA) and t-test. The results indicated that the IC50 level of cisplatin anti-cancer drug, as a common drug in the market, is significantly lower than Lagenaria siceraria extract. However, the extract of this plant revealed a significant growth inhibitory effect on lung cancer cells. The results also showed that Lagenaria siceraria extract is an effective cytotoxic compound on lung cancer cells. More extensive studies are needed to find effective plant extracts compounds to find and design new and effective cancer treatment drugs. Keywords: Lagenaria siceraria, Cell line, Lung cancer, IC50, MTTassay


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