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Nicotine is an alkaloid compound commonly found in tobacco smoke. This compound, as an agonist of nicotinic acetylcholine receptors (nAChRs), binds to these receptors and induces many downstream intracellular signaling pathways. Numerous studies have shown that nicotine leads to changes in the expression of microRNAs (miRs). Among these miRs, miR-124 is an essential post-transcriptional regulator of several cancer-related and inflammatory genes expression. The present study aimed to investigate the nicotine‑induced effects on miR-124 expression in colon cancer cell lines. First, the SW-480 colon carcinoma cell line was inoculated in 6-well plates to determine the effects of treatments with low and high concentrations of nicotine (1 and 10 μM) on the expression level of miR-124 and α7-subtype of nAChR (α7nAChR) and STAT-3 by quantitative real-time PCR. Next, the electroporation method was used to transfer specific siRNA targeting α7nAChR expression after identification of the optimum dose (60 nM) and time (48 h) of α7nAchR-siRNA. Following that, the changes in the expression of miR-124, α7nAChR, and STAT-3 were measured after transfection of the α7nAchR-siRNA in combination with nicotine treatment. The findings showed that nicotine increases the expression of both miR-124 and α7nAChR and decreases the STAT-3 mRNA expression. Transfection with α7nAChR-siRNA prevents the observed effects of nicotine which indicates that these effects of nicotine are α7nAChR dependent. Nicotine alters miR-124 expression in the SW-480 cell line and exerts its effects through the α7nAChR/miR-124/STAT-3 axis. Studying the exact molecular mechanism of nicotine signaling can be useful both in diagnosing and treatment of nicotine-related diseases and finding novel potential therapeutic benefits. [ABSTRACT FROM AUTHOR]

The natural ingredient-based therapy extracted from various plants is assumed more tolerable to cancer patients' bodies than synthetic drugs. To tell successful novel cancer therapy, an indicator is needed to confirm the result. VEGF is a pro-angiogenic factor that leads the cancer cells' invasion to another organ and is associated with high mortality in patients. Hence, the VEGF level is widely used as one of the successful cancer therapy indicators. Purposed of research elucidated the effect of Mandarin orange peel ethanol extract (EEKJM) to improve the reduction of VEGF levels in colon cancer in vitro. This study was an experimental posttest and control group design to analyze different level of VEGF on cancer cells at end of the study. Research began with the preparation of EEKJM followed by a cytotoxic extract test using the MTT assay method. Furthermore, the measurement of VEGF levels by the VEGF ELISA kit protocol. One Way ANOVA was performed to analyze data. Results showed the cytotoxic test of the EEKJM and doxorubicin against WiDr cells was 240 g/ml and 1.59 µg/ml. VEGF levels showed decreasing after being induced with EEKJM and doxorubicin with doses ½ IC50 and ¼ IC50. Conclusion: EEKJM was able to reduce VEGF levels and viability of WiDr colon cancer cells. The advantages are as much as initial evidence of the ability of the orange to treat cancer even though further animal studies or evidence of a pathway of decrease of the VEGF or other signals are required. [ABSTRACT FROM AUTHOR]

Abstract Objective Lipopolysaccharide (LPS) is a major cell wall component of gram-negative bacteria. Colon bacteria contribute to LPS which promotes colon cancer metastasis. The objective of this study was to survey the effect of LPS on cell viability and gene expression of 55 molecular targets in human colon cancer cells. Results LPS did not affect the viability of COLO 225 cells under the culture conditions but affected the expression of a number of genes important in inflammatory responses and cancer development. LPS increased TTP family, GLUT family and DGAT1 mRNA levels but decreased DGAT2a and DGAT2b expression in the human colon cancer cells. LPS also increased COX2, CXCL1, ELK1, ICAM1, TNFSF10 and ZFAND5 but decreased BCL2L1, CYP19A1 and E2F1 mRNA levels in the colon cancer cells. These data suggest that LPS has profound effects on gene expression in human colon cancer cells.

The study aimed to investigate antioxidant activities of two different thermally treated sesame (Sesamum indicum L.) leaf ethanol extract, steamed sesame leaf extract (SSLE) and roasted sesame leaf extract (RSLE), and their inhibitory effects on uncontrolled growth and increased metastatic properties in human colon cancer cell lines. Both SSLE and RSLE contained pedaliin as the major polyphenol and its aglycon, pedalitin, as a minor component and exhibited radical scavenging activities and ferric reducing antioxidant power. SSLE and RSLE decreased growth of HT29 and HCT116 colon cancer cells, which was attributed to the induction of apoptosis and cell cycle arrest at either G2/M (by SSLE in HCT116) or S phase (by RSLE in HCT116). Furthermore, SSLE and RSLE inhibited migration and adhesion in both cell lines. These results indicate that thermally treated sesame leaves retained pedaliin content and exhibited antioxidant activities and inhibitory activities against the growth and metastatic properties of colon cancer cells. [ABSTRACT FROM AUTHOR]

Objective: Lipopolysaccharide (LPS) is a major cell wall component of gram-negative bacteria. Colon bacteria contribute to LPS which promotes colon cancer metastasis. The objective of this study was to survey the effect of LPS on cell viability and gene expression of 55 molecular targets in human colon cancer cells. Results: LPS did not affect the viability of COLO 225 cells under the culture conditions but affected the expression of a number of genes important in inflammatory responses and cancer development. LPS increased TTP family, GLUT family and DGAT1 mRNA levels but decreased DGAT2a and DGAT2b expression in the human colon cancer cells. LPS also increased COX2, CXCL1, ELK1, ICAM1, TNFSF10 and ZFAND5 but decreased BCL2L1, CYP19A1 and E2F1 mRNA levels in the colon cancer cells. These data suggest that LPS has profound effects on gene expression in human colon cancer cells. [ABSTRACT FROM AUTHOR]

Cottonseed contains many bioactive molecules including plant polyphenols. Cottonseed value might be increased by providing high-value bioactive polyphenols for improving nutrition and health. However, there was a lack of molecular evidence for cottonseed bioactivity in mammalian cells. One widely used method for evaluating the bioactivity of natural products is quantitative real-time-PCR (qPCR). The selection of stably expressed internal reference genes is a crucial task of qPCR assay for data analysis. The rationale for reference gene selection is that a lower standard deviation of the cycle of threshold (Cq) among the treatments indicates a more stable expression of the gene. The objective of this study was to select reference genes in human colon cancer cells (COLO 205) treated with cottonseed-derived gossypol and bioactive extracts along with bacterial endotoxin lipopolysaccharides (LPS). SYBR Green qPCR was used to analyze the mRNA levels of a wide range of biomarkers involved in glucose transport, lipid biosynthesis, inflammatory response, and cancer development. qPCR data (10,560 Cq values) were generated from 55 genes analyzed from 64 treatments with triplicate per treatment for each gene. The data showed that B-cell lymphoma 2 (Bcl2) mRNA was the most stable among the 55 mRNAs analyzed in the human colon cancer cells. Glyceraldehyde 3 phosphate dehydrogenase (Gapdh) and ribosome protein L32 (Rpl32) mRNAs were not good qPCR references for the colon cancer cells. These observations were consistent regardless of the treatment comparison between gossypol and LPS, glanded and glandless seed extracts, seed coat and kernel extracts, or treatment for 8 and 24 h. These results suggest that Bcl2 is a preferable reference gene for qPCR assays in human colon cancer cells treated with cottonseed-derived gossypol and bioactive extracts as well as LPS. The extensive qPCR results firmly support the conclusion that the Bcl2 gene is stably expressed at the mRNA level in the human colon cancer cells regardless of the treatment, suggesting that Bcl2 gene expression is not regulated at the mRNA level but at the post-transcriptional level. These results should facilitate studies designated to evaluate bioactivity on gene expression regulation by cottonseed molecules and other natural and synthetic molecules for nutrition and health uses. [ABSTRACT FROM AUTHOR]

Existing studies on the biological activity of theabrownins are not based on their free state but on the complexes of theabrownins, polysaccharides, proteins, and flavonoids. In this study, theabrownins (TBs-C) were prepared by weak alkali oxidation of tea polyphenols. The ultraviolet-visible scanning spectrum of TBs-C showed two characteristic absorption peaks at 203 and 270 nm. The zeta potential of the TBs-C aqueous solution was negative, and the values varied from - 6.26 to -19.55 mV with a solution pH of 3-9. Storage conditions of pH 5.0-7.0 and around 25 °C were beneficial for the physical and chemical stability of the TBS-C solution. Cells were treated with series concentrations and examined by MTT, HE staining, PI immunofluorescence staining, flow cytometry, and real-time PCR to investigate the antiproliferative effect of TBs-C on human colon cancer HT-29 cells. The results showed that TBs-C, particularly at 500 µg/mL, inhibited cell growth. TBs-C induced HT-29 cell apoptosis, as confirmed by morphological changes, nucleus propidium iodide staining, and distributions of the cell cycle. The apoptotic mechanism may be due to the intracellular redox imbalance induced by TBs-C. [ABSTRACT FROM AUTHOR]

Theabrownins (TBs) are prepared from dark tea and contain a large number of complex heterogeneous components, such as carbohydrates, proteins, and flavonoids, which are difficult to remove. In addition, some toxic and harmful extraction solvents are used to purify TBs. These obstacles hinder the utilization and industrialization of TBs. In this study, tea polyphenols were used as substrates and polyphenol oxidase and sodium bicarbonate (NaHCO3) were used successively to prepare theabrownins (TBs-E). The UV-visible characteristic absorption peaks of the TBs-E were located at 203 and 270 nm and Fourier-transform IR analysis showed that they were polymerized phenolic substances containing the hydroxy and carboxyl groups. The TBs-E aqueous solution was negatively charged and the absolute values of the zeta potential increased with increasing pH. A storage experiment showed that TBs-E were more stable at pH 7.0 and in low-temperature environments around 25°C. HT-29 human colon cancer cells were used to evaluate the biological activity of TBs-E through 3-(4,5)-dimethylthiahiazo (-z-y1)-3,5-di- phenytetrazoliumromide (MTT), H&E staining, propidium iodide immunofluorescent staining, flow cytometry, and real-time PCR assays. The TBs-E significantly inhibited cell growth and caused late apoptosis, particularly at the dose of 500 μg/ml. The TBs-E markedly reduced the expression of antioxidant enzyme genes and increased the generation of reactive oxygen species to break the redox balance, which may have led to cell damage and death. These results will promote research and industrialization of TBs.

A lectin (designated as ARL) was purified first time from the Asparagus racemosus root with the molecular weight of 14.0 kDa containing about 4.8% carbohydrate. ARL showed hemagglutination activity in both mice and human erythrocytes that were inhibited by three complex sugars among the 26 sugars tested. ARL was thermostable that mostly preserved activity at its optimum pH 8.0. Around 48% and 52.5% human colorectal cancer (HCT-116) cells growth was inhibited by 160 μg/ml of ARL and 256 μg/ml of previously purified Geodorum densiflorum rhizome lectin (GDL). Induction of apoptosis in HCT-116 cells was confirmed by Hoechst 33342 staining, caspase inhibitors, but ROS generation was only observed for ARL. The expression level of BAX and p53 genes increased with a decrease of PARP gene expression for both lectins. The expression of FAS and FADD were increased with the decrease of WNT after treatment with GDL. ARL inhibited 68% and 26% of Ehrlich ascites carcinoma cell growth in vivo in mice after treating with 3.0 and 1.5 mg/kg/day doses for five consecutive days. ARL increased the expression level of NFκB and arrested S cell cycle phase in EAC cells, in contrast, G 2 /M phase was arrested by ARL and GDL in HCT-116. • ARL (Mr. 14 kDa), a thermostable glycoprotein was purified from A. racemosus root. • ARL & GDL inhibited HCT-116 cell growth and arrested G 2 /M cell cycle phase. • Apoptosis was confirmed by Hoechst-33342, caspase inhibitors and ROS for ARL. • The lectins altered several genes involved in multiple apoptotic pathways. • ARL inhibited EAC cells growth in vivo in mice and arrested S cell cycle phase. [ABSTRACT FROM AUTHOR]

Brassinin derived from Chinese cabbage has been reported to act as an anti-cancer agent on prostate, liver, and colon cancer cells. However, its mechanism and impact are largely unknown in colon cancer cells. Here, we first published a report that Brassinin induces apoptosis and inhibits the survival of colon cancer cells by activating p53. We found that Brassinin induces p53 and p21 dose- and time-dependent manner in wild type of p53 colon cancer cells. Interestingly, Brassinin induces apoptosis in wild-type of p53 cancer cells, but not in null-type of p53 cancer cells dose dependently. Additionally, Brassinin induces apoptosis through L5. Furthermore, Brassinin enhanced the apoptotic effect with doxorubicin by activating p53. Altogether, our findings suggest that Brassinin is a new p53 regulator via induce apoptosis and inhibit the proliferation in colon cancer cells. [ABSTRACT FROM AUTHOR]

KDM5c is a histone demethylase that specifically demethylates trimethylated and dimethylated H3 Lys-4 to play a central role in transcriptional repression. C-Jun is a proto-oncogene and promotes cell proliferation when ectopically accumulated, but can be ubiquitinated by SCF (FBXW7), leading to its degradation. FBXW7 is an E3 ubiquitin ligase of c-Jun, and exhibits carcinostasis in colon cancer. Here, we report that overexpression of KDM5c in human colon cancer cells results in attenuated FBXW7 transcription and accumulated c-Jun protein, leading to increased proliferation of colon cancer cells. We show that overexpression of KDM5c can result in increased c-Jun protein levels and decreased ubiquitin levels, with no significant change in mRNA levels of c-Jun. KDM5c overexpression blocks the ubiquitin-proteasome proteolytic pathway of c-Jun by down-regulating the expression of FBXW7. KDM5c down-regulation of FBXW7 occurs by demethylation of H3K4me3 at TSS and downstream of the FBXW7 gene. And interaction of KDM5c with H3K4me3 downstream of FBXW7 gene may be followed by recruitment of DNMT3b to methylate the spatially close CpG island located near the FBXW7 TSS. This methylation represses FBXW7 gene expression, which can reduce c-Jun degradation via the ubiquitin-proteasome pathway. TCGA database analysis revealed high expression of KDM5c in colon cancer tissues. KDM5c expression in colon cancer was correlated with poor overall survival of patients in the first 7 years. Data from TCGA showed that high expression of KDM5c was correlated with high DNA methylation of the FBXW7 gene, but was not positively correlated with methylation of the Jun gene. These results suggest that KDM5c regulation of colon cell proliferation is mainly mediated by the KDM5c-FBXW7-c-Jun axis.

Low density lipoprotein receptor related protein-1 (LRP-1) is a large ubiquitous endocytic receptor mediating the clearance of various molecules from the extracellular matrix. Several studies have shown that LRP-1 plays crucial roles during tumorigenesis functioning as a main signal pathway regulator, especially by interacting with other cell-surface receptors. Discoïdin Domain Receptors (DDRs), type I collagen receptors with tyrosine kinase activity, have previously been associated with tumor invasion and aggressiveness in diverse tumor environments. Here, we addressed whether it could exist functional interplays between LRP-1 and DDR1 to control colon carcinoma cell behavior in three-dimensional (3D) collagen matrices. We found that LRP-1 established tight molecular connections with DDR1 at the plasma membrane in colon cancer cells. In this tumor context, we provide evidence that LRP-1 regulates by endocytosis the cell surface levels of DDR1 expression. The LRP-1 mediated endocytosis of DDR1 increased cell proliferation by promoting cell cycle progression into S phase and decreasing apoptosis. In this study, we identified a new molecular way that controls the cell-surface expression of DDR1 and consequently the colon carcinoma cell proliferation and apoptosis and highlighted an additional mechanism by which LRP-1 carries out its sensor activity of the tumor microenvironment.

Yongyong Yang,1,* Shenyi Lian,2,* Lin Meng,1,* Zhihua Tian,3 Qin Feng,2 Yue Wang,2 Ping Wang,2 Like Qu,1 Chengchao Shou1 1Key Laboratory of Carcinogenesis and Translational Research (Ministry of Education/Beijing), Department of Biochemistry and Molecular Biology, Peking University Cancer Hospital & Institute, Beijing 100142, China; 2Department of Pathology, Peking University Cancer Hospital & Institute, Beijing 100142, China; 3Central Laboratory, Peking University Cancer Hospital & Institute, Beijing 100142, China *These authors contributed equally to this work Background: Phosphatase of regenerating liver-3 (PRL-3) has been shown to be highly expressed in various types of cancers and is related to poor prognosis. Our previous study showed that silencing of PRL-3 leads to increased reactive oxygen species (ROS). However, the mechanism of PRL-3 regulating ROS is not clear.Materials and methods: PRL-3 or Repressor activator protein 1 (RAP1) was knockdown in human colorectal cancer cell lines HCT116 and SW480. The mRNA level was measured by quantitative real-time (qRT)-PCR and the protein level was measured by western blot. ROS was detected by specific oxidationsensitive fluorescent probes. Cell cycle was analyzed through flow cytometry. Luciferase assay and chromatin immunoprecipitation (ChIP) were performed to investigate the regulation of RAP1 by PRL-3. Gene expression correlation was analyzed through an interactive web server. Statistical analysis was performed with SPSS software.Results: Knockdown of PRL-3 significantly increases mitochondrial superoxide anion, mitochondria membrane potential, and induces cell cycle arrest. Decreased PRL-3-induced mitochondrial superoxide anion accumulation is related to the downregulation of RAP1, which could also affect the level of mitochondria superoxide anion. PRL-3 regulates the expression of RAP1 through binding to the promoter of rap1 gene. PRL-3 could regulate the expression of peroxisome proliferator-activated receptor gamma coactivator 1 alpha (PGC-1α) through the mediation of RAP1. Both PRL-3 and RAP1 could regulate the expression of manganese superoxide dismutase 2 (SOD2) and the uncoupling protein 2 (UCP2), which may be related to PRL-3 suppression induced mitochondria superoxide anion.Conclusion: Our study presents the first evidence that PRL-3 is involved in the regulation of mitochondria superoxide anion as a transcriptional factor. Keywords: PRL-3, cell cycle, mitochondria, RAP1, PGC-1α, superoxide anion

Abstract Background Royal jelly (RJ), the exclusive food for the larva of queen honeybee, is regarded as the novel supplement to promote human health. The function of RJ may be attributed to its major and unique fatty acid, 10-hydroxy-2-decenoic acid (10-HDA). The current study investigated the anti-inflammory function of 10-HDA on human colon cancer cells, WiDr, as well as its effect on the growth of pathogenic bacterium. Methods The pro-inflammatory cytokines, receptor antagonist cytokine (IL-1ra) and nuclear factor-kappa B (NF-κB) in WiDr cells was analyzed by Enzyme-linked immunosorbent assay (ELISA) or western blot. The growth inhibition of 10-HDA on bacterium was evaluated by determination of minimal inhibitory concentrations (MIC) and minimal bactericide concentrations (MBC). Results The production of pro-inflammatory cytokines, Interleukin (IL)-8, IL-1β and tumor necrosis factor-alpha (TNF-α) in WiDr cells was modulated by 10-HDA. IL-8 were dramatically declined by 10-HDA at 3 mM, while IL-1β and TNF-α were significantly decreased. 10-HDA increased IL-1ra in a dose manner. NF-κB pathway is primarily in response to prototypical pro-inflammatory cytokines, and NF-κB was reduced after 10-HDA treatment. 10-HDA acted as potent bactericide against animal- or human-specific pathogens, including Staphylococcus aureus, Streptococcus alactolyticus, Staphylococcus intermedius B, Staphylococcus xylosus, Salmonella cholearasuis, Vibro parahaemolyticus and Escherichia coli (hemolytic). Conclusions The current study showed that in vitro 10-HDA from RJ exhibited anti-inflammatory activity in WiDr cells, as well as anti-bacterial activity against animal pathogens. 10-HDA showed its potential as anti-imflammtory agent and bactericide to benefit human gastrointestinal tract.

KDM5c is a histone demethylase that specifically demethylates trimethylated and dimethylated H3 Lys-4 to play a central role in transcriptional repression. C-Jun is a proto-oncogene and promotes cell proliferation when ectopically accumulated, but can be ubiquitinated by SCF (FBXW7), leading to its degradation. FBXW7 is an E3 ubiquitin ligase of c-Jun, and exhibits carcinostasis in colon cancer. Here, we report that overexpression of KDM5c in human colon cancer cells results in attenuated FBXW7 transcription and accumulated c-Jun protein, leading to increased proliferation of colon cancer cells. We show that overexpression of KDM5c can result in increased c-Jun protein levels and decreased ubiquitin levels, with no significant change in mRNA levels of c-Jun. KDM5c overexpression blocks the ubiquitin-proteasome proteolytic pathway of c-Jun by down-regulating the expression of FBXW7. KDM5c down-regulation of FBXW7 occurs by demethylation of H3K4me3 at TSS and downstream of the FBXW7 gene. And interaction of KDM5c with H3K4me3 downstream of FBXW7 gene may be followed by recruitment of DNMT3b to methylate the spatially close CpG island located near the FBXW7 TSS. This methylation represses FBXW7 gene expression, which can reduce c-Jun degradation via the ubiquitin-proteasome pathway. TCGA database analysis revealed high expression of KDM5c in colon cancer tissues. KDM5c expression in colon cancer was correlated with poor overall survival of patients in the first 7 years. Data from TCGA showed that high expression of KDM5c was correlated with high DNA methylation of the FBXW7 gene, but was not positively correlated with methylation of the Jun gene. These results suggest that KDM5c regulation of colon cell proliferation is mainly mediated by the KDM5c-FBXW7-c-Jun axis. [ABSTRACT FROM AUTHOR]

Two new mixed-ligand coordination polymers [Zn(bdc)(bib)]n (1, H2bdc = 1,3-dicarboxybenzene, bib = 1,3-bis(imidazol-1-yl)benzene) and [Zn(NH2-bdc)(bib)]n (2, NH2–H2bdc = 2-amino-1,4-dicarboxybenzene) are successfully prepared by the reaction of zinc nitrate, N-containing and dicarboxylate ligands under the same hydrothermal conditions. The anti-viability of compounds 1 and 2 is assessed with CCK-8 against SW480 colon cancer cells. To further explore the underlying mechanism of compounds, the cell apoptosis and ROS detection are performed in this experiment. [ABSTRACT FROM AUTHOR]

Brassinin derived from Chinese cabbage has been reported to act as an anti-cancer agent on prostate, liver, and colon cancer cells. However, its mechanism and impact are largely unknown in colon cancer cells. Here, we first published a report that Brassinin induces apoptosis and inhibits the survival of colon cancer cells by activating p53. We found that Brassinin induces p53 and p21 dose- and time-dependent manner in wild type of p53 colon cancer cells. Interestingly, Brassinin induces apoptosis in wild-type of p53 cancer cells, but not in null-type of p53 cancer cells dose dependently. Additionally, Brassinin induces apoptosis through L5. Furthermore, Brassinin enhanced the apoptotic effect with doxorubicin by activating p53. Altogether, our findings suggest that Brassinin is a new p53 regulator via induce apoptosis and inhibit the proliferation in colon cancer cells.

Abstract Background Genistein has been known to inhibit proliferation and induce apoptosis in several kinds of cancer cells. While knowledge of genistein in regulating epithelial mesenchymal transition (EMT) of colon cancer cells is unknown. Methods To investigate the effects and mechanisms of genistein on EMT of colon cancer cells, HT-29 cells were used and treated by genistein and TNF-α in this paper. EMT was determined by cell invasion assays using a transwell chamber and the expression changes of EMT-related markers were confirmed by RT–PCR, Western blotting, and immunofluorescence staining. Results Genistein inhibited cell migration at 200 μmol/L. Genistein reversed the EMT of colon cancer cells by upregulation of E-cadherin and downregulation of N-cadherin, accompanied by the suppression of EMT related makers, such as Snail2/slug, ZEB1, ZEB2, FOXC1, FOXC2 and TWIST1. Moreover, genistein can inhibit the expression of notch-1, p-NF-κB and NF-κB, while promote the expression of Bax/Bcl-2 and caspase-3 in HT-29 cells. Conclusion The present study demonstrated that genistein suppressed the migration of colon cancer cells by reversal the EMT via suppressing the Notch1/NF-κB/slug/E-cadherin pathway. Genistein may be developed as a potential antimetastasis agent to colon cancer.

An attempt has been made to investigate the role of Zaleya decandra on the survival of HCT colon cancer cells in vitro. The cell viability and apoptosis were assessed using MTT assay and acridine orange/ethidium bromide staining, respectively. Mitrochondrial membrane permeability was assessed using rhodamine-123 staining. The activities of caspases were evaluated using ELISA kits. Treatment with Z. decandra extract increased the production of ROS in HCT cells and impaired the mitochondrial activity as evidenced by decreased reduction of tetrazolium salt and enhanced mitochondrial membrane permeability. The onset of apoptosis was evident in cells treated with the extact in acridine orange/ethidium bromide staining and was associated with elevated levels of initiator and effector caspases in the conditioned medium. In conclusion, Z. decandra treatment promoted the apoptosis of HCT colon cancer cells. Video Clip of Methodology: 6 min 53 sec: Full Screen Alternate

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We investigated the anti‐cancer effects of ESC in human colon cancer LoVo cells. Cell counting assay results showed that ESC inhibited the proliferation of LoVo cells. Cell cycle arrest results showed that cell cycle was arrested during the G0/G1 phase in the ESC‐treated LoVo cells. Western blot results showed that the cell cycle inhibitory proteins p53, p27, and p21 were increased, and cyclin D1, the cell cycle progressive protein, was decreased. Sp1 is a transcription factor regulating cell proliferation, was decreased in the ESC‐treated LoVo cells. Annexin V/propidium iodide staining results showed that ESC induces apoptosis in LoVo cells. Western blot results showed that Bax, cleaved caspase ‐3, ‐7, ‐9, and poly(ADP‐ribose) polymerase, which are proapoptotic proteins, were increased and the antiapoptotic protein Bcl‐2 was decreased. Taken together, ESC induced apoptosis and has an anti‐cancer effect in LoVo cells. [ABSTRACT FROM AUTHOR]

Tae-Eon Kim,1–3,* Chang Gun Kim,1–3,* Jin Soo Kim,4 Songwan Jin,4 Sik Yoon,5 Hae-Rahn Bae,6 Jeong-Hwa Kim,7,8 Young Hun Jeong,7,8 Jong-Young Kwak1–3 1Department of Pharmacology, School of Medicine, 2Department of Biomedical Sciences, The Graduate School, Ajou University, Suwon, South Korea; 3Immune Network Pioneer Research Center, Ajou University Medical Center, Suwon, South Korea; 4Department of Mechanical Engineering, Korea Polytechnic University, Gyeonggi, South Korea; 5Department of Anatomy, School of Medicine, Pusan National University, Yangsan, South Korea; 6Department of Physiology, College of Medicine, Dong-A University, Busan, South Korea; 7School of Mechanical Engineering, 8Department of Mechanical Engineering, Graduate School, Kyungpook National University, Daegu, South Korea *These authors contributed equally to this work Abstract: An artificial three-dimensional (3D) culture system that mimics the tumor microenvironment in vitro is an essential tool for investigating the cross-talk between immune and cancer cells in tumors. In this study, we developed a 3D culture system using an electrospun poly(ε-caprolactone) (PCL) nanofibrous scaffold (NFS). A hybrid NFS containing an uninterrupted network of nano- and submicron-scale fibers (400 nm to 2 µm) was generated by deposition onto a stainless steel mesh instead of an aluminum plate. The hybrid NFS contained multiplanar pores in a 3D structure. Surface-seeded mouse CT26 colon cancer cells and bone marrow-derived dendritic cells (BM-DCs) were able to infiltrate the hybrid NFS within several hours. BM-DCs cultured on PCL nanofibers showed a baseline inactive form, and lipopolysaccharide (LPS)-activated BM-DCs showed increased expression of CD86 and major histocompatibility complex Class II. Actin and phosphorylated FAK were enriched where unstimulated and LPS-stimulated BM-DCs contacted the fibers in the 3D hybrid NFS. When BM-DCs were cocultured with mitoxantrone-treated CT26 cells in a 3D hybrid NFS, BM-DCs sprouted cytoplasm to, migrated to, synapsed with, and engulfed mitoxantrone-treated CT26 cancer cells, which were similar to the naturally occurring cross-talk between these two types of cells. The 3D hybrid NFS developed here provides a 3D structure for coculture of cancer and immune cells. Keywords: 3D cell culture, electrospinning, nanofibrous scaffold, dendritic cell, colon cancer cell

Abstract Previous researches have reported that microcystin-LR (MC-LR) contributes to the progression of multiple types of carcinomas including colon cancer; however, the underlying molecular mechanisms remain unclear and require in-depth investigation. Here, the colon cell line DLD-1 was arranged for the analysis by the microRNA microarray which was associated with the cancer metastasis after MC-LR exposure. 31 human microRNAs were differentially expressed, including miR-221, which targeted 3′-UTR of PTEN mRNA and PTEN level was down-regulated by MC-LR treatment. Besides, MC-LR also induced the phosphorylation of STAT3, which can be reversed by adding miR-221 inhibitor and PTEN expression plasmid. Furthermore, miR-221 inhibitor, STAT3 siRNA and PTEN expression plasmid could reverse the effects of MC-LR induced migration with the accumulation of β-catenin in nuclei. In conclusion, our study suggested that MC-LR promoted the progression of colon carcinoma, at least in part, by regulating the expression miR-221, PTEN and STAT3 phosphorylation, which offers a novel perspective to understand the connection between MC-LR and colon cancer. Graphical abstract fx1 Highlights • MC-LR inhibits PTEN expression by up-regulating the level of miR-221. • MC-LR induces the phosphorylation of STAT3, which can be reversed by miR-221/PTEN. • PTEN and STAT3 suppress MC-LR-induced accumulation of β-catenin in nuclei. • MiR-221/PTEN and STAT3 signal pathway regulates the MC-LR-induced migration. [ABSTRACT FROM AUTHOR]

Doxorubicin (DOX) is a cytotoxic compound capable of instigating apoptosis in cancer cells. TP53 apoptosis effector (PERP) is a key mediator of apoptosis in multiple cell types. PERP transcription is activated by a range of pro-apoptotic stimuli. In the present study, we investigated the regulation of DOX-induced PERP transcription in colon cancer cells (SW480) by the transcriptional modulator myocardin-related transcription factor A (MRTF-A). We report that DOX treatment up-regulated MRTF-A expression paralleling PERP activation. DOX also promoted nuclear translocation of MRTF-A. On the contrary, MRTF-A depletion or inhibition attenuated DOX-induced apoptosis as evidenced by the MTT assay and caspase 3 cleavage. In accordance, MRTF-A depletion or inhibition dampened PERP transcription. Chromatin immunoprecipitation (ChIP) assay showed that DOX treatment promoted the binding of MRTF-A on the PERP promoter. Mechanistically, MRTF-A was recruited to the PERP promoter by activator protein 1 (AP-1). AP-1 interacted and cooperated with MRTF-A to activate PERP transcription. AP-1 silencing weakened PERP trans-activation by DOX presumably by compromising MRTF-A recruitment to the PERP promoter. In conclusion, our data suggest that MRTF-A might be a key regulator of DOX-induced PERP transcription in colon cancer cells. [ABSTRACT FROM AUTHOR]

Background: Royal jelly (RJ), the exclusive food for the larva of queen honeybee, is regarded as the novel supplement to promote human health. The function of RJ may be attributed to its major and unique fatty acid, 10-hydroxy-2-decenoic acid (10-HDA). The current study investigated the anti-inflammory function of 10-HDA on human colon cancer cells, WiDr, as well as its effect on the growth of pathogenic bacterium. Methods: The pro-inflammatory cytokines, receptor antagonist cytokine (IL-1ra) and nuclear factor-kappa B (NF-κB) in WiDr cells was analyzed by Enzyme-linked immunosorbent assay (ELISA) or western blot. The growth inhibition of 10-HDA on bacterium was evaluated by determination of minimal inhibitory concentrations (MIC) and minimal bactericide concentrations (MBC). Results: The production of pro-inflammatory cytokines, Interleukin (IL)-8, IL-1β and tumor necrosis factor-alpha (TNF-α) in WiDr cells was modulated by 10-HDA. IL-8 were dramatically declined by 10-HDA at 3 mM, while IL-1β and TNF-α were significantly decreased. 10-HDA increased IL-1ra in a dose manner. NF-κB pathway is primarily in response to prototypical pro-inflammatory cytokines, and NF-κB was reduced after 10-HDA treatment. 10-HDA acted as potent bactericide against animal- or human-specific pathogens, including Staphylococcus aureus, Streptococcus alactolyticus, Staphylococcus intermedius B, Staphylococcus xylosus, Salmonella cholearasuis, Vibro parahaemolyticus and Escherichia coli (hemolytic). Conclusions: The current study showed that in vitro 10-HDA from RJ exhibited anti-inflammatory activity in WiDr cells, as well as anti-bacterial activity against animal pathogens. 10-HDA showed its potential as anti-imflammtory agent and bactericide to benefit human gastrointestinal tract. [ABSTRACT FROM AUTHOR]

Glycan metabolism balance is critical for cell prosperity, and macromolecule glycosylation is essential for cell communication, signaling and survival. Thus, glycotherapy may be a potential cancer treatment. The aim of the present study was to determine whether combined synthetic glycoconjugates (GCs) induce changes in gene expression that alter the survival of colon cancer cells. The current study evaluated the effect of the GCs N‑acetyl‑D‑glucosamine modified polyamidoamine dendrimer and calix[4]arene scaffold on cancer cell proliferation, apoptosis, invasion and sensitivity to immune cell‑mediated killing. Using reverse transcription‑quantitative polymerase chain reaction, the expression of genes involved in the aforementioned processes was measured. It was determined that GCs reduce the expression of the glucosaminyltransferases Mgat3 and Mgat5 responsible for surface glycosylation and employed components of the Wnt signaling pathway Wnt2B and Wnt9B. In addition, the calix[4]arene‑based GC reduced cell colony formation; this was accompanied by the downregulation of the metalloproteinase Mmp3. By contrast, the dendrimer‑based GC affected the expression of the glucose transporter components Sglt1 and Egfr1. Therefore, to the best of our knowledge, the present study is the first to reveal that N‑acetyl‑D‑glucosamine‑dendrimer/calix[4]arene GCs alter mRNA expression in a comprehensive way, resulting in the reduced malignant phenotype of the colon cancer cell line HT‑29. [ABSTRACT FROM AUTHOR]

Previously, carnosic acid and carnosol have demonstrated anti-proliferative activity against different types of cancer. To obtain extracts enriched in these two key phenolic compounds, two different processes have been developed in the present work based on the use of two-step sequential supercritical fluid extraction (SFE). By removing the interfering, less active fractions in a first step (150 or 300 bar, 40°C, neat CO2, 60 min), suitable enrichment is achieved in the second step (150 bar, 40°C, CO2 + 7% ethanol, 120 min), and this leads to carnosic acid concentrations in the extract as high as 40% of total dry weight, which are among the highest concentrations that have been described with this type of process. The enriched extracts were tested against the HT-29 human adenocarcinoma cell line, showing enhancement of their antiproliferative activity by approximately 3-fold compared to previously reported SFE rosemary extracts and higher inhibitory effects at lower concentrations (30 µg mL−1 of extract). Thus, the proposed two-step SFE process effectively improves the carnosic acid and carnosol recovery in shorter processing times (180 min vs. 300 min). Moreover, the obtained extracts possess higher anti-proliferative activity and consume less solvent.

Evidence demonstrate that p53 mutations and microRNAs (miRs) are important components of 5-FU resistance in colorectal cancer (CRC). miR-338-3p has been reported associated with cancer prognosis. However whether or not it influences chemotherapy sensitivity and the underlying mechanisms have not been elucidated. Here, three types of human colon cancer cell lines, HT29 (mutant p53), HCT116 (wild-type p53), and HCT116 p53 -/- (deficient p53), were treated with 5-FU. We showed that expression of miR-338-3p was correlated with apoptosis and 5-FU resistance in colon cancer cells. Ectopic expression of miR-338-3p conferred resistance to 5-FU in HCT116 cells. Further experiments indicated that miR-338-3p mediated 5-FU resistance through down-regulation of mTOR expression. Moreover, inhibition of miR-338-3p in HT29 and HCT116 p53 -/- cells increased their sensitivity to 5-FU treatment. Furthermore, we detected autophagy changes in our experiment because mTOR was known prominently regulating autophagy and the competition between autophagy and apoptosis in response to 5-FU was a mechanism influencing 5-FU sensitivity. Our results reveal a critical and novel role of miR-338-3p in the correlation of 5-FU resistance with p53 status. Moreover, the miR-338-3p inhibitor has the potential to overcome 5-FU resistance in p53 mutant colon cancer cells. [ABSTRACT FROM AUTHOR]

Objective To observe the effect of down-regulated CDX2 gene on the migration and invasion abilities of colon cancer cells (SW480 and HT29) and investigate the role and mechanisms of CDX2 gene in occurrence and development of colon cancer metastasis. Methods CDX2 gene in HT29 and SW480 cells was down-regulated using lentivirus RNA interference (RNAi) vector. The interference efficiency of CDX2 was detected by qRT-PCR and Western blotting. The effect of down-regulated CDX2 expression on colon cancer cells' migration and invasion was determined by Transwell and wound heal methods. Then the effects of down-regulated CDX2 on the expressions of epithelial-mesenchymal transition (EMT)-related genes (E-cadherin, ZEB-1, Vimentin, Twist and Snail) were detected by RT-PCR and Western blotting. Results The constructed CDX2 siRNA expression vector could significantly inhibit the expression of CDX2 in HT29 and SW480 cells. Compared with those of the cells transfected with empty vector (LV-NT-shRNA) and non-transfected cells, the migration and invasion abilities of cells transfected with LV-CDX2-shRNA were markedly enhanced (P<0.05). E-cadherin expression was reduced while expressions of ZEB-1, Vimentin, Twist, and Snail were significantly increased (all P<0.05). Conclusion Down-regulating the expression of CDX2 can induce the occurrence of EMT, thus enhancing the invasion and migration of colon cancer cells. [ABSTRACT FROM AUTHOR]

Context:Shikonin is a kind of naphthoquinone compound found mainly in Lithospermum erythrorhizon Sieb,et Zucc. Previous studies have shown that Shikonin has anti-tumor, anti-inflammatory and extensive pharmacological effects. According to new studies, Shikonin could also modulate the immune system function, but the effect to NK (nature killer) cells is yet unknown. Objective:To investigate the effect and mechanism of Shikonin on NK cells proliferation and cytotoxicity to colon cancer cell line (Caco-2). Methods:The proliferation and cytotoxicity of NK cells cultured with Shikonin were detected with CCK-8 assay. The expressions of perforin, GranB and IFN-γ were examined with FCM. The content of TNF-alpha was disclosed with ELISA kit. p-ERK1/2 and p-Akt expression of NK cells were detected with western blot. Results:With CCK-8 assay, it is found that Shikonin could significantly enhance NK cells proliferation and cytotoxicity to colon cancer cells. With FCM assay, it is found that Shikonin could improve the expression of perforin and GranB in a dose-dependent manner. Shikonin had no effect on TNF-alpha and IFN-γ expression. In mechanism, the study shows that Shikonin could enhance the expression of p-ERK1/2 and p-Akt. Conclusions:Shikonin enhances NK cells proliferation and cytotoxicity via the improvement of perforin, GranB, p-ERK1/2 and p-Akt expression. [ABSTRACT FROM PUBLISHER]