Human Stomach Cancer Research Articles

Evaluation of the circadian rhythms of activity in mice implanted with human stomach cancer cell lines.

Evaluation of the circadian rhythms of activity in mice implanted with human stomach cancer cell lines.

Inhibiting the JNK/ERK signaling pathway with geraniol for attenuating the proliferation of human gastric adenocarcinoma AGS cells.

Geraniol, a natural compound found in the essential oils of various aromatic plants, has attracted attention for its probable anticancer effects. The molecular mechanisms of the cell proliferation suppression and apoptosis induction via geraniol in gastric cancer cells (AGS), however, remain unclear. Gastric cancer cells were treated with geraniol, and it was found that the IC50 values were 25 μM/ml, as determined by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. Results showed that 20 and 25 μM geraniol-induced reactive oxygen species (ROS) production (2'-7'dichlorofluorescin diacetate staining) and decreased mitochondrial membrane potential (rhodamine 123 staining) in AGS cells. Then, it effectively inhibited cell growth and induced apoptosis, confirmed through acridine orange/ethidium bromide, 4',6-diamidino-2-phenylindole, and propidium iodidestaining and molecular marker analysis in AGS cells. Also, geraniol potently diminished caspase-9, Bax, Bcl-2, and caspase-3 expression in AGS cells. We also evaluated the essential mechanism of the cytotoxic effect of geraniol. Moreover, the present study depicted that geraniol-induced cell death through mitochondrial ROS production and inhibited the phosphorylation form of mitogen-activated protein kinase (p38, MAPK, JNK, and ERK1/2) signaling pathway. Taken together, these results concluded that geraniol has a novel therapeutic property against human stomach cancer.

Antioxidant and Cytotoxic Activities of Beach Morning Glory (Ipomoea pes-caprae)

The present study aimed to evaluate the in-vitro and in-vivo antitumor potential of Ipomoea pes-caprae (I. pes-caprae). Petroleum ether (60-80oC), methanol, aqueous, and swaras extracts prepared from the whole plant of I. pes-caprae. Antioxidant activity of I. pes-caprae was evaluated by using 2, 2-diphenyl-1-picrylhydrazylhydrate (DPPH), reducing power, and hydroxyl ion scavenging assay methods. The cytotoxic effect of I. pes-caprae was evaluated by using Trypan blue and 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay against animal melanoma cancer (B16F10), human stomach cancer (Kato-III), and human colorectal cancer (HT-29) cell lines. The ultraviolet (UV), infrared (IR), nuclear magnetic resonance (NMR), and Mass spectra were recorded for the structure elucidation of isolated compounds. These isolated compounds of I. pes-caprae were evaluated biologically at two doses of 25 and 12.5 mg/kg body weight on male C57BL mice to explore the antitumor activity against melanoma cancer. The scavenging range (IC50) of the extracts of I. pes-caprae was found to be 63.40±2.26 to 74.77±2.28 μg/ml, and the cytotoxic range (CTC50) for MTT assay was found to be 62.54±3.27 to 73.70±1.32 μg/ml. Based on IR, NMR, and mass spectroscopic analysis and by chemical transformation, structures of three compounds A, B, and C were elucidated as 3’-methoxy-3,4’,5,7-tetrahydroxy flavone, 3,7-dimethoxy-8-methyl-4’,5-dihydroxy flavone, and 3’-methoxy-4’,5,7-trihydroxy flavone-3-glucoside, respectively. All isolated compounds were shown antitumor potential against melanoma cancer.

Biogenic and eco-benign synthesis of silver nanoparticles using jujube core extract and its performance in catalytic and pharmaceutical applications: Removal of industrial contaminants and in-vitro antibacterial and anticancer activities

Biosynthesis of nanoparticles has been rapidly developed in various fields, due to their broad spectrum of applications in the fields of environmental, pharmacology, and medicine. In this study, facile, rapid, eco-friendly and cost-effective method was used to green synthesis of silver nanoparticles (AgNPs) using jujube core extract (AgNPs- JCE ) and then used as antibacterial, anticancer and catalytic agents. The reaction parameters such as AgNO 3 concentration (1, 5, 10, 20 and 40 mM), reaction time (5, 30 and 60 min) and pH (without pH, 10 and 12) were discussed and optimized. The surface plasmon resonance peak at about 420 nm in the UV–Vis absorption spectrum confirmed the green synthesis of silver nanoparticles. Microscopic results revealed that the synthesized AgNPs- JCE were spherical in morphology with a size range of 25–35 nm.In addition, the subject AgNPs promising catalytic properties in the degradation of pollutants including rhodamine b (RhB) and eriochrome black T (EBT) as cationic and anionic contaminant under UV and visible light irradiations. The photocatalyst (AgNPs- JCE ) exhibited the degradation of 90.9 % and 84.7% for RhB and EBT contaminants after 80 min under UV irradiation, respectively. The antibacterial activities of AgNPs- JCE was checked against E. coli as Gram-positive bacteria and K. pneumoniae and S. aureus as Gram-negative bacteria with MIC and MBC values of (1.26 and 1.26 μ g/ml), (2.5 and 2.5 μ g/ml), and (2.5 and 10 μ g/ml), respectively. Finally, the cytotoxicity of synthesized nanoparticles against AGS as human stomach cancer cell line was determined at several concentrations (2.5, 5, 10, 20, 50, and 100 μ g/ml) using MTT assay. Finding of this research suggested the suitability of AgNPs- JCE as pollutants degradation, antibacterial and anticancer drug development. • First report of biogenic silver nanoparticles synthesis using jujube core extract. • Green synthesized of AgNPs was analyzed by UV/Vis, FT-IR, XRD, EDS, FESEM and TEM. • Exhibited potential photocatalytic degradation in rhodamine b (RhB) and eriochrome black T (EBT) under both UV and sunlight irradiations. • Biosynthesized AgNPs showed good antibacterial activity against gram-positive and gram-negative bacteria. • AgNPs exhibited excellent anticancer activity against AGS as human stomach cancer cell line.

Near-Infrared Ratiometric Two-Photon Probe for pH Measurement in Human Stomach Cancer Tissue.

Stomach cancer is a global health issue because of its incidence and mortality rates worldwide. We developed a near-infrared (NIR) emissive ratiometric two-photon (TP) probe (HCC1) for the quantitative analysis of pH in live cells and human stomach tissues. The probe design is based on a restrained hemicyanine core that controls the intramolecular charge transfer from 2-naphthol, with a suitable pKa value (7.50) under physiological conditions. The probe exhibited improved quantum yield, stability, and TP activity under physiological conditions. In addition, intracellular pH titration (pH 4.0 to 10.0) of HCC1 revealed an ideal intracellular pKa of approximately 7.2, negligible cytotoxicity, and TP excited fluorescence in situ, thereby allowing direct imaging of the cellular pH in live cells and tissues. Ratiometric two-photon microscope imaging with HCC1 of human stomach tissue revealed a clear intratissue pH variation among normal, adenoma, and cancer tissues. Our results demonstrate that HCC1 is useful as an NIR imaging probe for in situ pH-related studies and in cancer research.

Potentially Bioaccessible Phenolics from Mung Bean and Adzuki Bean Sprouts Enriched with Probiotic-Antioxidant Properties and Effect on the Motility and Survival of AGS Human Gastric Carcinoma Cells.

Gastric digests from mung (MBS) and adzuki (ABS) bean sprouts enriched with probiotic Lactobacillus plantarum 299v were tested for their antioxidant potential, as well as antiproliferative and antimotility properties, in human stomach cancer cells (AGS). The digest of ABS contained quercetin and kaempferol derivates, while kaempferol and apigenin derivates were dominant in MBS. Compared to the controls, the probiotic-rich sprouts had a higher antioxidant potential—by 13% and 9%, respectively. Adzuki bean sprouts decreased the viability of AGS already at low concentrations (25% motility inhibitions). MBS and ABS displayed dose-independent cytostatic effects. The ABS extracts decreased the proliferation of AGS more effectively than the MBS extracts—0.2‰ ABS exerted c.a. 70% of inhibitions. Moreover, the phytochemicals from the probiotic-rich sprouts considerably reduced this activity. The increased vinculin level, the apoptotic shape of cell nuclei, and the reduced cell motility and proliferation indicate that the extracts exhibited cytostatic and cytotoxic activity.

Improved Method for Obtaining of Arctigenin from Arctium Lappa L. and its Antiproliferative Effect on Human Hepatocarcinoma HepG2 Cells

Background: Arctigenin is a lignan found in Arctium lappa L. (Asteraceae) that displays anti-inflammatory activities. Previous studies showed that the crude extract of A. Lappa has antitumor activity in human liver carcinoma, lung and stomach cancer cells. The aim of this study was to obtain arctigenin from A. lappa L., as well as to evaluate its antiproliferative effects in cells of liver carcinoma (HepG2) and fibroblasts (NIH/3T3). Methods: Arctigenin was obtained from the hydrolysis of arctiin, which was isolated from the crude extract of A. lappa. The effects of arctigenin and arctiin on HepG2 cell viability and cell adhesion were analyzed by MTT method. Adhesion assay was also carried out to evaluate the antitumor activity. Results: Our results showed that the analytical process to obtain arctigenin was fast and easy. In vitro experiments showed that arctigenin (107-269 μM) decreased HepG2 cells viability and did not cause cytotoxicity on NIH/3T3 cells. Arctigenin (27-269 μM) demonstrated anti-adhesion in HepG2 cells in a concentration-dependent manner, when compared with control. Conclusion: These results suggest a promising pharmacological activity for arctigenin as an antiproliferative compound.

Antiproliferative Effects of Ocimum basilicum Methanolic Extract and Fractions, Oleanolic Acid and 3-epi-Ursolic Acid

Aims: The aim of the current study was to identify active compound(s) responsible for the antiproliferative effects of O. basilicum and explore their underlying mechanism/s. Background: Plants have been the source of medicines for the treatment of various diseases since ancient times. Ocimum basilicum (Sweet Basil, Bobai Tulsi) has been used in the folk medicine for the treatment of human liver, spleen and stomach cancers. Background: Plants have been the source of medicines for the treatment of various diseases since ancient times. Ocimum basilicum (Sweet Basil, Bobai Tulsi) has been used in the folk medicine for the treatment of human liver, spleen and stomach cancers. Objective: To emphasize the importance of O. basilicum as a potential novel non-toxic alternative to the conventional anticancer therapy. Method: O. basilicum (aerial parts) methanolic extract and fractions were screened against HT-144, MCF-7, NCI-H460 and SF-268 human cancer cell lines using sulforhodamine B assay. The more active Petroleum Ether Insoluble (PEI) fraction was fractionated into six sub-fractions (OB-1 to OB-6). Four pure compounds (3-O-methyl ursolic acid, oleanolic acid, 3-epi-ursolic acid and ursolic acid) were isolated from the more potent sub-fraction OB- 6. Triple channel immunofluorescence microscopy was employed to observe the effects of methanolic extract, PEI fraction, sub-fractions OB-5 and OB-6, 3-epi-ursolic acid and oleanolic acid on the cytoskeleton and nuclei of MCF-7 cells. Result: The methanolic extract and the PEI fraction exhibited selectively greater growth inhibition against MCF-7 cell line (TGI: 56 and 36.2 µg/ml, respectively). By using triple channel immunofluorescence microscopy, it was observed that the methanolic extract, PEI fraction, sub-fraction OB-5 and 3-epi-ursolic acid induced irregular mitotic spindle formation and slowing of mitotic progression in MCF-7 cells while sub-fraction OB-6 induced mitotic arrest in the prophase stage. F-actin aggregation was also visible in PEI fraction, subfraction OB-5 and 3-epi-ursolic acid treated MCF-7 cells. Conclusion: These results emphasize the importance of O. basilicum as a potential novel non-toxic alternative to the conventional anticancer therapy and suggest that it inhibits the growth of MCF-7 cancer cells via multiple mechanisms such as interaction with the microtubules and mitotic spindle apparatus, and F-actin aggregation.

New chalcone-type compounds and 2-pyrazoline derivatives: synthesis and caspase-dependent anticancer activity.

Aim: There is a continuous and urgent need for new anticancer agents with novel structures and target selectivity. Methods & results: The anticancer activity of the prepared compounds was assessed against human lung (A549) and stomach (AGS) cancer cell lines and evaluated in the noncancer human lung fibroblast (MRC-5) cell line. 2-Pyrazolines were devoid of toxicity in all cell lines used, chalcones bearing a β-(benz)imidazole moiety being toxic toward AGS cell line. Mechanistic studies showed that these compounds trigger loss of cell viability and mitochondrial membrane potential, while eliciting morphological traits compatible with regulated cell death, which was ultimately shown to derive from caspase activation, specifically caspase-3. Conclusion: Chalcones 1-3 have been identified as new and promising anticancer agents toward the AGS cell line.

A fibronectin-coated gold nanostructure composite for electrochemical detection of effects of curcumin-carrying nanoliposomes on human stomach cancer cells.

Curcumin, which is produced by the medicinal herbaceous plant Curcuma longa, has been widely investigated for use as a potential anticancer drug. In this study, the potential toxicity of curcumin-carrying nanoliposomes (curcumin-NLC) toward human stomach cancer cells (MKN-28) was investigated using a new cell-based electrochemical sensing platform. To satisfy both biocompatibility and electroconductivity of the electrodes, the density of the gold nanostructure and the coating conditions of extracellular matrix proteins (fibronectin and collagen) were optimized. The developed platform enabled the successful adhesion and long-term growth of stomach cancer cells on the chip surface, allowing label-free and real-time monitoring of cell viability in a quantitative manner. According to the electrochemical results, both bare curcumin and curcumin-NLC showed toxicity toward MKN-28 cells in the concentration range of 10-100 μM, which was consistent with the results obtained from a conventional colorimetric method (CCK-8). Remarkably, at a low concentration range (<50 μM), this electrochemical platform determined the decrease in cell viability to be approximately 22.8%, 33.9% and 53.1% in the presence of 10, 30, and 50 μM of curcumin-NLC, respectively, compared with the 1.3%, 18.5%, and 28.1% determined by CCK-8, making it 1.7-2 times more sensitive than the conventional colorimetric assay. Hence, it can be concluded that the newly developed fibronectin-coated electroconductive platform is highly promising as an electrochemical detection tool for the sensitive and precise assessment of the anticancer effects of various food-derived compounds with low toxicity.

自発運動はヒト胃がん細胞株85As2誘発性がん悪液質に関連する心機能障害を改善する

自発運動はヒト胃がん細胞株85As2誘発性がん悪液質に関連する心機能障害を改善する

Study of refractive index of human stomach cancer tissue in THz frequency range

In this paper, the samples of healthy and cancer tissues of the internal and external sides of a human stomach were studied by time-domain terahertz reflectometry in the frequency range from 0.2 to 1 THz. Since healthy and cancer tissues have different morphological structure, and also contain different amounts of water, their optical properties and spectral characteristics differ. It was found that the refractive index of internal side of stomach cancer tissue is greater than for the healthy one. The refractive index of the external side of stomach cancer tissue is less than for the healthy one. This effect should be taken into account for development of stomach cancer diagnosis.

Co-expression of Interleukin-17A molecular adjuvant and prophylactic Helicobacter pylori genetic vaccine could cause sterile immunity in Treg suppressed mice

The increasing clinical significance of Helicobacter pylori (H. pylori) in human stomach cancer has led to global efforts to eradicate this pathogen. Recent studies have confirmed the importance of some cytokines such as Interleukin-18 (IL-18), Interleukin-8 (IL-8), Interleukin-17A (IL-17A) and Interleukin-22 (IL-22) in the pathogenesis of the so-called bacterium. This study was designed to compare the effects of Type 1T helper (Th1), Type 2T helper (Th2) cells, Regulatory T cells (Treg) and T helper 17 (Th17) modulatory effects on the efficacy of designed H. pylori vaccine by incorporating some molecular adjuvants in Treg competent and Treg suppressed groups. A bicistronic vector was used for simultaneous expression of codon-optimized Outer inflammatory protein a (OipA) gene and modified mice IL-18, IL-17A, IL-22 and Foxp3 (forkhead box P3) cytokines from four cassettes. Immunization of mice groups was performed using produced plasmids intradermally. Specific IgG1 and IgG2 and IgA antibody titers produced in mice were confirmed by enzyme-linked immunosorbent assay (ELISA) in sera and intestine obtained four weeks after the last immunization. After being stimulated with a mixture of both anti-CD28 mAb and H. pylori lysate, frequencies of single Interferon-Gamma (IFN-γ), single IL-17 and dual IFN-γ/IL-17-secreting T-cells were documented using dual-color FluoroSpot. The kinetics of Th1, Th2 and Th17 in the immunized animals was determined by relative quantification of IL-17A, IL-22, IFN-γ, IL-8, IL-2 and IL-4 specific mRNAs. Four weeks after bacterial challenge, quantitative colony count in the isolated and homogenized stomachs was utilized to assess the level of protective immunity among all groups. The results of immunologic assays showed that the highest cell-mediated immunity cytokines were produced in IL-17 receiving group in which the Treg responses were suppressed previously by the administration of the Foxp3 as an immunogen. In addition, potent clearance of Helicobacter pylori infection was seen in this group as well.

Classification of human stomach cancer using morphological feature analysis from optical coherence tomography images

Optical coherence tomography is radiation-free, and it is considered a tool of optical biopsy. Classification of normal and cancerous tissues is very important for the guidance of surgeons. Here, we develop the morphological feature analysis-based classification (MFAC) method, combining it with machine learning to identify cancerous tissues. We extract five quantitative morphological features from one OCT image through the structured analysis. Five classifiers are involved to make a classification: the support vector machine, the K-nearest neighbor, the random forest, logic regression, and the conventional threshold method. Sensitivity, specificity, and accuracy are used to evaluate these classifiers and are compared with each other. We launched the experimental research of the imaging of ex vivo patients’ stomach cancerous tissue with the OCT system. The results showed the three additional features specially designed for stomach cancer are remarkably better than the traditional image feature. The best feature demonstrated over 95% accuracy under all five classifiers. The designed feature based on the layer structure of the stomach tissue is significantly effective. This MFAC method will be used to image the in vivo tissue in clinical applications in the future.

Benzoquinones from Cyperus spp. trigger IRE1α-independent and PERK-dependent ER stress in human stomach cancer cells and are novel proteasome inhibitors

BackgroundThe roots and tubers of several species of the Cyperus genus are used in several parts of the world as foodstuffs and beverages. The genus is rich in several classes of quinones, however their biological properties have not been studied before. PurposeWe evaluated the anticancer effect of several benzoquinones isolated from the genus and described their mechanism of action towards cancer cells. MethodsThe most potent molecules were selected according to their effect upon cell viability. The mechanism of cell death was studied by using pharmacological inhibitors of caspases, caspase-3/4/9 activity assays, annexin-V/7-AAD by flow cytometry and intracellular reactive oxygen species and calcium levels through fluorescence spectroscopy. Elucidation of the involvement of distinct branches of the ER stress pathway was pursued by RT-PCR and WB for mRNA and protein expression levels, respectively, as well as pharmacological inhibitors. Proteasome inhibitory activity was assessed by using purified 20S catalytic subunit with the fluorogenic substrate Suc-Leu-Leu-Val-Tyr-AMC. ResultsCytotoxicity studies against cancer cell lines showed that the human gastric cancer cell line AGS was the most susceptible, the most potent molecule, hydroxycyperaquinone, exhibiting an IC50 close to 1 µM. Morphological and biochemical traits suggested that a process of regulated cell death was taking place, which was shown to be intrinsic pathway-independent. Results indicated that benzoquinones exert their toxicity by triggering ER stress, as shown by increased expression of CHOP (mRNA and protein levels), intracellular reactive oxygen species, changes in calcium dynamics and caspase-4 activation. Proteasome inhibition by these molecules is described for the first time. ConclusionHydroxycyperaquinone is a novel sub-micromolar inhibitor of the 20S catalytic core of the 26S proteasome, causing cell death via IRE1α-independent/PERK-dependent pathways in stomach cancer cells. Its presence in products consumed orally may be of relevance for gastric tumors.

Investigation of The Effect of Radiosensitization of Tungsten Oxide Nanoparticles on AGS Cell Line of Human Stomach Cancer in Megavoltage Photons Radiation

Radiotherapy which consists of external beam radiotherapy (EBRT) and internal radioisotope therapy (RIT), has a wide usage for treating cancer as clinical trials.This study provides some conditions to prove that tungsten oxide nanoparticles (WO3) is a radiosensitizer.The peripheral blood mononuclear cells (PBMCs) were used to calculate inhibitory concentration (IC).The AGS cell line exposed the concentration of 89.6 µg/mL (IC20) nanoparticle WO3 that was optimal and its radiosensitization was examined in megavoltage photons radiation of 6 MV x-rays. The sensitivity enhancement ratio (SER) and dose enhancement factor (DEF) was determined 1.24 and 1.68 respectively. We described the mechanisms of creating nanoparticles WO3 toxicity and genotoxicity in different concentrations on AGS cell line. The mean size of WO3 NPs by transmission electron microscopy wasmeasured 31.89 3.82 nm.Tungsten oxide Nanoparticles cause to reduce cell viability, remove membrane and damage to DNA. There was a meaningful increasing in damages to DNA and proliferation cell potency and also significantly reducing cell viability in concentrations more than 100 µg/mL.

Three new inositol derivatives from Chisocheton paniculatus

Three new inositol derivatives (1–3) were isolated from the leaves of Chisocheton paniculatus collected in Vietnam, along with four known compounds, including a limonoid (4), two triterpenoids (5–6), and a tocopherol (7). The structures of the new compounds were elucidated by 1D- and 2D-NMR and HRESIMS analyses. Compound 4 showed the highest cytotoxicities against the human lung cancer A549 and cervical cancer HeLa cell lines, with IC50 values of 7.3 and 8.8 µM, respectively, among the isolated compounds. Compounds 5 and 7 displayed moderate to weak cytotoxicities against the A549, HeLa, and human stomach cancer GSU cell lines, with IC50 values ranging from 17.7 to 68.0 µM.

The Role of YAP Oncogene in Metastasis and Mechano‐medicine

YAP is a WW domain‐containing protein that acts as a potent oncogene and stemness factor. It is one of the two main effectors of the Hippo tumor suppressor pathway. YAP is a transforming gene of the chromosome 11q22 amplicon, and its expression is elevated at high frequency in human cancers, including liver, breast, ovarian and stomach cancer. Our recent work showed that YAP regulates the acto‐myosin network by suppressing Rho‐GTPase via Rho‐GTPase activating protein 29 (ARHGAP29) being a direct transcriptional target of YAP in human gastric cancer. We showed that YAP promoted the expression of ARHGAP29 to suppress the RhoA‐LIMK‐cofilin pathway, thereby destabilizing F‐actin. The overexpression of YAP caused cytoskeletal rearrangement by altering the dynamics of F‐actin/G‐actin turnover, thus promoting migration. In a mouse model, circulating tumor cells (CTCs) exhibited an increase in ARHGAP29 RNA level compared with cells at primary tumor sites. Moreover, increased ARHGAP29 expression correlated with shortened survival of human gastric cancer patients. Importantly, we showed that ARHGAP29 is critical in regulating cancer metastasis in a mouse model of liver cancer metastasizing to lungs.We have also identified YAP and TAZ specific transcripts and we are characterizing them now as mechano‐responsive transcripts that regulate cell metastasis and mechano‐sensing.Cancer cells are generally softer than normal cells. By increasing the rigidity of cancer cells to the level of normal cells via novel therapeutic interventions, and by reducing rigidity of extracellular matrix of tumors we could provide a new modality to treat cancer in unison with other standard therapies.Support or Funding InformationStart‐Up Funds from National University of Singapore (R‐185‐000‐2710‐733) and Mechanobiology Institute (R‐714‐018‐006‐271)This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.

Integrated transcriptome and in vitro analysis revealed anti-proliferative effect of citral in human stomach cancer through apoptosis

Cancer is the second leading cause of death globally, particularly stomach cancer is third most common causes of cancer death worldwide. Citral possesses anti-tumor activity in various cancer cell lines, However its effect toward stomach cancer and its mechanism of action is have yet to be elucidated. The goal of the present study is to elucidate the role of citral in stomach cancer using transcriptome and in vitro approaches. We performed transcriptome analysis using RNA-seq and explored its capability to persuade apoptosis in AGS human stomach cancer cell lines in vitro. Furthermore, the enrichment and KEGG pathway results suggested that there are several genes involved to induce apoptosis pathway. Furthermore, our study also demonstrated that citral arrested colony formation and migration of cancer cells significantly than that of untreated cells. RNA-seq revealed a total of 125 million trimmed reads obtained from both control and citral treated groups respectively. A total number of 612 differentially expressed genes (DEGs) were identified which includes 216 genes up-regulated and 396 genes down-regulated genes after treatment. The enrichment analysis identified DEGs genes from transcriptome libraries including cell death, cell cycle, apoptosis and cell growth. The present study showed the significant inhibition effect upon citral by regulating various genes involved in signaling pathways, inhibits metastasis, colony formation and induced apoptosis both in silico and in vitro.

Generation of Cancer Cell Clones to Visualize Telomeric Repeat-containing RNA TERRA Expressed from a Single Telomere in Living Cells.

Telomeres are transcribed, giving rise to telomeric repeat-containing long noncoding RNAs (TERRA), which have been proposed to play important roles in telomere biology, including heterochromatin formation and telomere length homeostasis. Recent findings revealed that TERRA molecules also interact with internal chromosomal regions to regulate gene expression in mouse embryonic stem (ES) cells. In line with this evidence, RNA fluorescence in situ hybridization (RNA-FISH) analyses have shown that only a subset of TERRA transcripts localize at chromosome ends. A better understanding of the dynamics of TERRA molecules will help define their function and mechanisms of action. Here, we describe a method to label and visualize single-telomere TERRA transcripts in cancer cells using the MS2-GFP system. To this aim, we present a protocol to generate stable clones, using the AGS human stomach cancer cell line, containing MS2 sequences integrated at a single subtelomere. Transcription of TERRA from the MS2-tagged telomere results in the expression of MS2-tagged TERRA molecules that are visualized by live-cell fluorescence microscopy upon co-expression of a MS2 RNA-binding protein fused to GFP (MS2-GFP). This approach enables researchers to study the dynamics of single-telomere TERRA molecules in cancer cells, and it can be applied to other cell lines.