microRNA-133b Regulates Cell Proliferation and Cell Cycle Progression via Targeting HuR in Colorectal Cancer

MicroRNAs (miRNAs/miRs) have been identified to serve a key role in the development of tumors. However, the role of miR-133b in colorectal cancer (CRC) remains largely unclear. This study will investigate the role and mechanism of miR-133b in CRC. Reverse transcription-quantitative polymerase chain reaction analysis was performed to detect the level of miR-133b in CRC cell lines. Bioinformatics software TargetScan predicted the potential target genes of miR-133b, and a dual luciferase reporter assay was used to confirm this. To investigate the role of miR-133b in CRC cells, miR-133b was upregulated or downregulated in CRC cell lines (SW620 and HT-29) by transfecting with a miR-133b mimic or inhibitor, respectively. Subsequently, cell viability was analyzed using MTT assay, whereas cell apoptosis and the cell cycle distribution were analyzed by flow cytometry. In addition, the associated protein levels were detected using western blot analysis. The results demonstrated that miR-133b was significantly downregulated in CRC cell lines when compared with the normal colonic epithelial NCM-460 cell line. Human antigen R (HuR; also termed ELAVL1) was demonstrated to be a direct target of miR-133b and was negatively regulated by miR-133b. HuR was also notably upregulated in the CRC cell lines when compared with the normal control. Transfection of SW620 and HT-29 cells with the miR-133b mimic significantly inhibited cell viability, and induced cell apoptosis and G1 phase arrest, while upregulation of HuR demonstrated the opposite effects. Furthermore, the present data demonstrated that the miR-133b mimic significantly enhanced the protein levels of p21 and p27, and downregulated cyclin D1 and cyclin A levels in SW620 and HT-29 cells; the opposite effects were observed following treatment with the miR-133b inhibitor. In conclusion, the data indicate that miR-133b suppressed CRC cell growth by targeting HuR.

Protective effect of two Thai pigmented rice cultivars against H2O2-induced oxidative damage in HT-29 cell culture

Radicals derived from exogenous and endogenous sources are considered to be the principal cause of genetic damage. Exogenous and endogenous radicals participate in the reactive oxygen species (ROS) formation, which leads to damages in the DNA, RNA, proteins and lipids. However, dietary compounds, mainly from pigmented rice, are an essential source of antioxidants that help protect cells from damage. This study seeks to determine the antioxidant properties and cytoprotective effect of two Thai pigmented rice extracts namely the glutinous black rice (native name: Neaw dum moa37) and red rice (native name: Hom gradung-nga57) on H2O2-induced damage in HT-29 cells. The bioactive compound contents, as well as antioxidant activities of both rice extracts, were investigated. The protective effect of rice extracts on H2O2-induced damage was executed following the co-incubation method. HT-29 cells were exposed to H2O2 and different rice extract concentrations for 3 h and an MTT assay was used to measure the viability of the cell. The ROS level was determined using the 2′,7′-dichlorofluorescin diacetate (DCFDA). The result showed that glutinous black rice extract contained significantly higher contents of all analysed antioxidants and activities than red rice extract. Glutinous black rice showed a higher cytotoxic effect compared to red rice. At the non-toxic concentration of both rice extracts, the HT-29 cells were guarded against the H2O2 induced oxidative stress. Besides, the intracellular ROS accumulation result from H2O2 exposure was significantly reduced in the presence of rice extracts for both glutinous black rice and red rice compared to control. Hence, this study has demonstrated the potential properties of both pigmented rice extracts in alleviating H2O2-mediated damage in HT-29 cells.

Mechanism of Pterostilbene-Induced Cell Death in HT-29 Colon Cancer Cells

Pterostilbene is a dietary phytochemical that has been found to possess several biological activities, such as antioxidant and anti-inflammatory. Recent studies have shown that it exhibits the hallmark characteristics of an anticancer agent. The aim of the study was to investigate the anticancer activity of pterostilbene against HT-29 human colon cancer cells, focusing on its influence on cell growth, differentiation, and the ability of this stilbene to induce cell death. To clarify the mechanism of pterostilbene activity against colon cancer cells, changes in the expression of several genes and proteins that are directly related to cell proliferation, signal transduction pathways, apoptosis, and autophagy were also evaluated. Cell growth and proliferation of cells exposed to pterostilbene (5–100 µM) were determined by SRB and BRDU assays. Flow cytometric analyses were used for cell cycle progression. Further molecular investigations were performed using quantitative real-time RT-PCR. The expression of the signaling proteins studied was determined by the ELISA method. The results revealed that pterostilbene inhibited proliferation and induced the death of HT-29 colon cancer cells. Pterostilbene, depending on concentration, caused inhibition of proliferation, G1 cell arrest, and/or triggered apoptosis in HT-29 cells. These effects were mediated by the down-regulation of the STAT3 and AKT kinase pathways. It may be concluded that pterostilbene could be considered as a potential therapeutic option in the treatment of colon cancer in the future.

Synthesis and anti-tumor activity evaluation of salinomycin C20-O-alkyl/benzyl oximes derivatives

Seventeen C20-O-alkyl/benzyl oximes derivatives were synthesized in concise and effective method. Most of these derivatives showed tens to several hundreds nanomolar IC50 values against HT-29 colorectal, HGC-27 gastric and MDA-MB-231...

Antiproliferative Effect of Epilobium parviflorum Extracts on Colorectal Cancer Cell Line HT-29

The Epilobium species, rich in various active phytochemicals, have been widely used in folk medicine to treat several diseases including benign prostatic hyperplasia. Despite being demonstrated on some type of cancer cells such as prostate cancer, their potential anti-cancerous role on colorectal adenocarcinoma cells has not been studied yet. According to the World Health Organization (WHO), colon cancer is the third most common form of cancer, resulting over 800 000 deaths every year worldwide. The present study demonstrates the anti-cancerous activity of aqueous and ethanolic Epilobium parviflorum extracts in colon cancer cell line HT-29 cells in vitro. The both type of extracts reduced the cell viability of HT-29 cells in a dose dependent manner. Gene expression analysis of HT-29 cells demonstrated an increase at apoptotic genes, caspase 3 and caspase 8. Nuclear fragmentation of apoptotic cells was also demonstrated through TUNEL assay as well as immunostaining experiments. On the other hand, same lethal concentrations of E. parviflorum extracts were not profound on non-cancerous human fibroblast cell line BJ cells. Our results indicate that E. parviflorum may also be used as a therapeutic agent against colon cancers.