Normal Human Colonic Epithelial Cells Research Articles

HmiR-34c-3p upregulation inhibits the proliferation of colon cancer cells by targeting EIF3D.

Our study desired to investigate how miR-34c-3p regulates colon cancer cell proliferation and what is the relationship between miR-34c-3p and EIF3D. HCEpiC (normal human colonic epithelial cells), SW620, HT-29, SW480, and HCT-116 (human colon cancer cells lines) were used in our study. SW620 cells were chosen and divided into blank, miR-34c-3p mimics, miR-34c-3p NC, miR-34c-3p inhibitors, Lv-EIF3D, Lv-NC, and miR-34c-3p mimics+Lv-EIF3D groups. qRT-PCR was used for the detection of miR-34c-3p and EIF3D mRNA expressions. Dual-luciferase reporter assay was performed to investigate the effect of miR-34c-3p on EIF3D. Western blot was performed to detect EIF3D, cyclin D1, and c-Myc expressions. Clone formation and MTT assay were used to measure cell proliferation ability. colon cancer cells had lower miR-34c-3p expression, but higher EIF3D expression compared with HCEpiC. EIF3D mRNA expression was regulated negatively by miR-34c-3p. In the miR-34c-3p mimics group, colon cancer cell proliferation was significantly decreased, whereas c-Myc and cyclin D1 expressions were downregulated. Colon cancer cell proliferation in miR-34c-3p inhibitors and Lv-EIF3D groups was enhanced, and c-Myc and cyclin D1 expressions were decreased. The results suggested that by targeting EIF3D, miR-34c-3p inhibited colon cancer cell proliferation.

Imaging of human cells exposed to an antifungal antibiotic amphotericin B reveals the mechanisms associated with the drug toxicity and cell defence

Amphotericin B is an antibiotic used in pharmacotherapy of life-threatening mycotic infections. Unfortunately, the applicability of this antibiotic is associated with highly toxic side effects. In order to understand molecular mechanisms underlying toxicity of amphotericin B to patients, two cell lines, human normal colon epithelial cells (CCD 841 CoTr) and human colon adenocarcinoma cells (HT-29) were cultured in the presence of the drug and imaged with the application of fluorescence lifetime imaging microscopy and Raman scattering microscopy. The results of the cell viability assays confirm high toxicity of amphotericin B towards human cells. The images recorded demonstrate effective binding of amphotericin B to biomembranes. Analysis of the images reveals the operation of a defence mechanism based upon the elimination of molecules of the drug from living cells via formation of small amphotericin B-containing lipid vesicles. The fact that exosomes formed are devoid of cholesterol, as concluded on the basis of the results of Raman analysis, suggests that sequestration of sterols from the lipid phase of biomembranes is not a sole mechanism responsible for the toxic side effects of amphotericin B. Alternatively, the results imply that molecules of the drug present directly within the hydrophobic membrane core disturb the lipid membrane structure and affect their biological functions.

Biological activity of oxygenated pinene derivatives on human colon normal and carcinoma cells

AbstractThe antitumour activity of the oxygenated derivatives of pinene is still poorly understood. We hypothesized that the derivatives of pinene, (−)‐myrtenol, (−)‐myrtanol, and (−)‐myrtenal, would express different effects on human colon tumour cells and normal intestinal cells in vitro. For cytotoxic and nitric oxide (NOx) assays, human colon tumour (HT29) and human normal colon epithelial cells (CCD 841 CoTr) were incubated for 24 h with 0.1–5.0 mM of the terpenoids. The NOx level was determined in cells pre‐incubated or not with lipopolysaccharide (LPS). All three terpenoids decreased the activity of succinate dehydrogenase in tumour and normal cells, with a mixture (1 : 1 : 1) of the pinene derivatives having the strongest effect (with half‐maximal inhibitory concentrations, IC50, of 2.56 and 1.05 mM, respectively). The cytotoxic effect of the terpenes on tumour cells was observed at concentrations higher than 2 mM, with (–)‐myrtenal exerting the strongest activity (IC50 = 5.3 mM). Normal cells were more sensitive to the activity of the terpenes, especially when administered as a mixture (IC50 = 0.08 mM). The terpenoids synergistically stimulated tumour cells for NOx release and acted antagonistically in the case of normal cells. Tumour cells released the highest (1.111 μM) and normal cells released the lowest (0.146 μM) quantities of NOx after incubation with 1 mM terpenoid mixture. LPS‐induced tumour cells produced higher quantities of NOx than the unstimulated culture. An opposite effect was observed for normal intestinal cells. The investigated pinene derivatives inhibit the activity of human colon tumour cells in vitro by affecting mitochondrial enzyme activity, NOx release, and membrane stability. Differences in the bioactivity of the individual compounds can be determined on the basis of their structure and degree of oxidation.

MiR-522-3p Promotes Tumorigenesis in Human Colorectal Cancer via Targeting Bloom Syndrome Protein.

miR-522-3p is known to degrade bloom syndrome protein (BLM) and enhance expression of other proto-oncogenes, leading to tumorigenesis. This study aimed to investigate the molecular mechanisms of miR-522-3p in human colorectal cancer (CRC) cells. Expressions of miR-522-3p in CRC and adjacent tissues, as well as in normal human colon epithelial cell line (FHC) and five CRC cell lines, were detected. Human CRC cell lines, HCT-116 and HT29, were transfected with miR-522-3p mimic, inhibitor, or scrambled controls. Then cell viability, apoptosis, cell cycle progression, and the expressions of c-myc, cyclin E, CDK2, and BLM were assessed. It was found that miR-522-3p was highly expressed in CRC tissues when compared to adjacent nontumor tissues and was highly expressed in CRC cell lines when compared to FHC cells. miR-522-3p overexpression promoted cell viability, reduced apoptotic cell rate, arrested more cells in the S phase, and upregulated c-myc, cyclin E, and CDK2 expression. BLM was a target gene of miR-522-3p, and miR-522-3p suppression did not exert antiproliferative and proapoptotic activities when BLM was silenced. These findings demonstrate that miR-522-3p upregulation negatively regulates the expression of BLM, with upregulation of c-myc, CDK2, and cyclin E, and thereby promoting the proliferation of human CRC cells.

The expression profile and clinical application potential of hsa_circ_0000711 in colorectal cancer.

IntroductionCircular RNAs, as a class of long-time-ignored non-coding RNA, have been revealed as multifunctional RNAs in recent years, especially in the cancer research. However, the mechanism of most circular RNAs and their clinical application values in human cancers remain unknown, including in colorectal cancer (CRC).MethodsIn this study, we focused on the expression pattern and clinical values of hsa_circ_0000711 in CRC. The expression level of hsa_circ_0000711 in 101 paired CRC tissues and 3 CRC cell lines (HCT116, COLO205, and H-T29), as well as human normal colon epithelial cell line NCM460, were measured by quantitative real-time polymerase chain reaction.ResultsOur results revealed that the expression level of hsa_circ_0000711 was significantly downregulated in CRC tissues (P=9.35E-16) and CRC cell lines (P<0.01). In addition, the area under the receiver characteristic curves was 0.81. The sensitivity and specificity were 0.91 and 0.58, respectively. Meanwhile, our study showed that low expression of hsa_circ_0000711 could serve as an independent prediction biomarker associated with poor overall survival of CRC patients (hazard ratio =2.409; 95% CI: 1.276–4.547; P=0.004).ConclusionAll of these results indicated that hsa_circ_0000711 may play a crucial role in CRC carcinogenesis and could be a potential effective biomarker for the diagnosis and prognosis of CRC.

Migration inhibition of water stress proteins from Nostoc commune Vauch. via activation of autophagy in DLD-1 cells

Water stress proteins (WSP1) from Nostoc commune Vauch. had been proven to selectively induce colon cancer cells apoptosis. In this study, the effect of WSP1 on migration of human colon cancer cells was investigated. It showed that WSP1 inhibited DLD-1 cell migration, but with an insignificant effect on normal human colon epithelial cells. The data further indicated that WSP1 activated autophagy through down regulation of PI3K/AKT/mTOR pathway. Meanwhile, β‑catenin was degraded by autophagy, which then restrained epithelial-mesenchymal transition (EMT) of DLD-1 cell and its migration was subsequently suppressed significantly. The same changes occurred in xenografted nude mice according to the obtained immunohistochemical results. Consistently, the application of autophagy inhibitor largely reversed the inhibited migration by WSP1 treatment. Taken together, WSP1 could suppress migration of DLD-1 cells by autophagy inhibited EMT. The results suggested that WSP1 possessed the potential as a selective therapeutic agent against metastatic colon cancer.

Abstract 488: Epigenetic regulation of colorectal cancer stem cells by the miR-221/QKI5 axis

Abstract Colorectal cancer stem cells (CSCs) have extensive abilities to initiate tumor formation, and are responsible for the progression and metastasis of colorectal cancers. MicroRNAs (miRNAs) are non-coding RNAs of 18-24 nucleotides that post-transcriptionally regulate expression of multiple genes by targeting their 3'untranslated regions (UTRs). miRNAs regulate various biological processes, and function as important regulators of CSC properties. miR-221 is one of the oncogenic miRNAs frequently upregulated in human cancers including colorectal cancer. Previous studies have shown that miR-221 enhances proliferation, migration, and invasion of cancer cells. However, the role of miR-221 in human colorectal CSCs is not fully elucidated. In this study, we analyzed the colorectal CSCs directly isolated from the surgical specimens of colorectal cancer patients to explore the role of miR-221 in CSCs. Expression profile of 754 miRNAs in the CD44+/EpCAM+ CSC and non-tumorigenic cancer cell (NTC) populations was analyzed by multiplex semi-quantitative PCR. Prognostic impact of miR-221 expression in CSCs was analyzed using the TCGA database. Target genes of miR-221 were validated by luciferase reporter assays and Western blot experiments. The roles of miR-221 and its target gene QKI-5 in colorectal CSCs were evaluated by organoid and xenotransplantation assays. Comparison of the expression levels of 754 miRNAs in the CSCs and NTCs resulted in the identification of 10 miRNAs upregulated or downregulated in CSCs compared to NTCs. Among them, miR-221 was most highly expressed in CSCs and its expression level was very low in NTCs and human normal colon epithelial cells of human colorectal cancer specimens (n=6, p&amp;lt;0.05). The estimated 5-year overall survival rate for miR-221high patients (n=177) was significantly lower than miR-221low ones (n=116) in TCGA dataset (54.6% vs. 73.6%; p&amp;lt;0.001). The TNM stages at diagnosis were not significantly associated with the expression level of miR-221. In a multivariate analysis, miR-221 expression were significantly associated with overall survival (p=0.009). miR-221 targeted an RNA binding protein (RBP) QKI-5 and suppressed its expression. Knockdown of miR-221 in human colorectal cancer patient-derived xenograft cells suppressed the organoid formation in vitro in a QKI-5-suppression-dependent manner, and tumor formation initiated by colorectal CSCs in vivo (n=6, p=0.015 ). Our results suggest that the miR-221/QKI5 axis plays important roles in the regulation of CSC properties in human colorectal cancers. Because miR-221 was preferentially and highly upregulated in human colorectal CSCs, but not in normal stem/progenitor cells, it is possible that miR-221 will be a biomarker that reflects amount and/or activity of CSCs in colorectal cancers and an attractive target to selectively attach colorectal CSCs by avoiding damages on their normal counterparts. Citation Format: Junko Mukohyama, Yohei Shimono, Piero Dalerba, Taichi Isobe, Qingjiang Hu, Debashis Sahoo, Naoki Shibuya, Hironobu Minami, Koshi Mimori, Yoshihiro Kakeji, Akira Suzuki. Epigenetic regulation of colorectal cancer stem cells by the miR-221/QKI5 axis [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 488.

Synthesis and Binding Behaviour of New Isomers of Bis-Thiourea

The two new symmetrical bis-thiourea compounds, 2,2’-[{(terephthaloylbis(azanediyl)bis (carbonothioyl)bis(azanediyl)}bis(4-methylpentanoicacid)] 1A and 2,2’-[{(isophthaloylbis (azanediyl)bis(carbonothioyl)bis(azanediyl)}bis(4-methylpentanoic acid)] 1B have been synthesized by reacting terephthaloyl/isophthaloyl chloride and L-leucine in high yields. Newly synthesized bis-thiourea derivatives were characterized using FTIR, 1D NMR, 2D NMR, MS spectrometry and Elemental analysis. Their binding properties with various cations were also carried out using UV-vis titration experiments. Both isomers exhibited effective binding against Ag+, Cu2+, Ni2+, Hg2+, Pb2+, Fe2+, and Fe3+ in the presence of other cations, such as Na+, Co2+, Cd2+, Zn2+, Mn2+, Mg2+, Ca2+, Sn2+, and Al3+. Stoichiometries of the host-guest complexation for both isomers were found to be 1:4 by plotting molar-ratio curves. Pearson Product moment correlation coefficient was in the range of 0.83-0.99 and nonlinear regression equation was used to calculate dissociation constant (Kd). Both compounds displayed weak antibacterial activities against gram-positive and gram negative bacteria. Cytotoxicity testing on CCD841 Normal human colon epithelial cell line showed that both compounds are non-toxic, with an IC50 value of 1.50 mg/mL.

Novel polypyridyl ruthenium complexes acting as high affinity DNA intercalators, potent transcription inhibitors and antitumor reagents

Six novel polypyridyl ruthenium complexes with (E)-2-styryl-1H- imidazo[4,5-f][1,10]phenanthroline ligand and its analogues have been designed to enhance the DNA intercalation ability of their model compound [Ru(bpy)2(pip)]2+ (bpy = 2,2′-bipyridine, pip = 2-phenyl-1H-imidazo[4,5-f][1,10]phenanthroline). As shown in the optimized geometry of the complexes, the introduction of styryl group not only extended the conjugated area of the intercalative ligand, but also retained the excellent planarity. These two merits have been proven to be beneficial for their DNA intercalation, thus greatly improved their inhibition activity towards DNA transcription by RNA polymerase and DNA topoisomerase, two enzymes closely related to both DNA and tumor cell growth. The relationships between the substituent group structures and the biological activities have also been investigated from energetic and electronic aspects by quantum chemistry calculations. Results from cell cytotoxicity and apoptosis assay testified that the styryl substituted ruthenium complexes possessed higher antitumor activity than [Ru(bpy)2(pip)]2+, as expected. As quantified in the MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay, the tumor cell death is caused mostly through apoptosis for Ru2 and Ru3, while non-apoptotic processes for Ru1, Ru4 and Ru5. In vitro fluorescence evaluation revealed that all complexes located mainly in cytoplasm, but the three complexes with high antiproliferative activity could enter nucleus. All complexes have shown apparent lower cytotoxicity towards normal human colon epithelial cell CCD-841-CON than the examined tumor cell lines.

Differences in the Effects of EGCG on Chromosomal Stability and Cell Growth between Normal and Colon Cancer Cells.

The tea catechin epigallocatechin-3-gallate (EGCG) proved to be the most potent physiologically active tea compound in vitro. It possesses antioxidant as well as pro-oxidant properties. EGCG has the effect of inducing apoptosis of tumor cells and inhibiting cell proliferation. Whether this effect is associated with the antioxidant or pro-oxidative effects of EGCG affecting the genome stability of normal and cancer cells has not been confirmed. Here, we selected Human normal colon epithelial cells NCM460 and colon adenocarcinoma cells COLO205 to investigate the effects of EGCG (0–40 μg/mL) on the genome stability and cell growth status. Chromosomal instability (CIN), nuclear division index (NDI), and apoptosis was measured by cytokinesis-block micronucleus assay (CBMN), and the expression of core genes in mismatch repair (hMLMLH1 and hMSH2) was examined by RT-qPCR. We found that EGCG significantly reduced CIN and apoptosis rate of NCM460 at all concentrations (5–40 μg/mL) and treatment time, EGCG at 5 μg/mL promoted cell division; EGCG could significantly induce chromosome instability in COLO205 cells and trigger apoptosis and inhibition of cell division. These results suggest that EGCG exhibits different genetic and cytological effects in normal and colon cancer cells.

Biphasic regulation of spindle assembly checkpoint by low and high concentrations of resveratrol leads to the opposite effect on chromosomal instability

Resveratrol (RSV) is a naturally occurring polyphenolic phytoalexin possessing numerous health-promoting effects. Chromosomal instability (CIN), usually results from defective spindle assembly checkpoint (SAC), is a major contributor to many diseases. While it’s recently recognized that RSV exhibits a nonlinear dose response for disease prevention, whether it’s the case for its role in CIN remains unknown. Here, we investigated the potential of a broad range of RSV concentrations (0.01–100μM) on CIN and the underlying mechanisms in human normal colon epithelial NCM460 cells. CIN was measured by cytokinesis-block micronucleus assay; mitotic fidelity was determined by aberrant mitosis analysis; SAC activity was assessed by nocodazole-challenge assay, and the expression of SAC genes was examined by RT-qPCR. We found that 0.1μM RSV significantly reduced CIN (P<0.01), while 100μM RSV significantly induced it (P<0.05). Mitotic infidelity was significantly prevented by 0.1μM RSV but promoted by 100μM RSV (P<0.05 for both). Moreover, the function of SAC was sustained and impaired by 0.1μM and 100μM RSV, respectively. Several SAC genes, including Aurora-B, Aurora-C, Plk-1 and CENP-E, were significantly up-regulated and down-regulated by 0.1μM and 100μM RSV, respectively (P<0.05). In conclusion, RSV exhibited a biphasic dose-dependent effect on CIN that was exerted via the regulation of mitotic fidelity through the SAC network. The health implications of these findings were summarized.

Perilla frutescens Extracts Protects against Dextran Sulfate Sodium-Induced Murine Colitis: NF-κB, STAT3, and Nrf2 as Putative Targets.

Perilla frutescens is a culinary and medicinal herb which has a strong anti-inflammatory and antioxidative effects. In the present study, we investigated the effects of Perilla frutescens extract (PE) against dextran sulfate sodium (DSS)-induced mouse colitis, an animal model that mimics human inflammatory bowel disease (IBD). Five-week-old male ICR mice were treated with a daily dose of PE (20 or 100 mg/kg, p.o.) for 1 week, followed by administration of 3% DSS in double distilled drinking water and PE by gavage for another week. DSS-induced colitis was characterized by body weight loss, colon length shortening, diarrhea and bloody stool, and these symptoms were significantly ameliorated by PE treatment. PE administration suppressed DSS-induced expression of proinflammatory enzymes, including cyclooxygenase-2 and inducible nitric oxide synthase as well as cyclin D1, in a dose-dependent fashion. Nuclear factor-kappa B (NF-κB) and signal transducer and activator of transcription 3 (STAT3) are major transcriptional regulators of inflammatory signaling. PE administration significantly inhibited the activation of both NF-κB and STAT3 induced by DSS, while it elevated the accumulation of Nrf2 and heme oxygenase-1 in the colon. In another experiment, treatment of CCD841CoN human normal colon epithelial cells with PE (10 mg/ml) resulted in the attenuation of the tumor necrosis factor-α-induced expression/activation of mediators of proinflammatory signaling. The above results indicate that PE has a preventive potential for use in the management of IBD.

Different molecular organization of two carotenoids, lutein and zeaxanthin, in human colon epithelial cells and colon adenocarcinoma cells

Two cell lines, human normal colon epithelial cells (CCD 841 CoTr) and human colon adenocarcinoma cells (HT-29) were cultured in the presence of exogenous carotenoids, either zeaxanthin or lutein. Both carotenoids demonstrated cytotoxicity with respect to cancer cells but not to normal cells. Cells from both the cell lines were analyzed with application of fluorescence lifetime imaging microscopy and Raman scattering microscopy. Both imaging techniques show effective incorporation of carotenoid molecules into growing cells. Comparison of the Raman scattering and fluorescence lifetime characteristics reveals different molecular organization of carotenoids in the carcinoma and normal cells. The main difference consists in a carotenoid aggregation level which is substantially lower in the carcinoma cells as compared to the normal cells. Different molecular organization of carotenoids was interpreted in terms of a different metabolism of normal and carcinoma cells and has been concluded to provide a possibility of cancer diagnosis based on spectroscopic analyses.

Evaluation of anticancer activity of water and juice extracts of young Hordeum vulgare in human cancer cell lines HT-29 and A549.

Introduction and objective. Barley (Hordeum vulgare L.) is known as a rich source of different bioactive compounds. At present, considerable attention of researchers is focused on young barley grass. It can be a good source of dietary minerals, vitamins, carbohydrates, amino acids, phenolic compounds and proteins. It is possible that the composition of chemical ingredients beneficial for health may induce an anticancer potential of young barley in human colon adenocarcinoma (HT-29) and human lung adenocarcinoma (A549) cell lines. Materials and method. Hordeum vulgare water extract (HWE) and Hordeum vulgare juice extract (HJE) were prepared. Cell proliferation and viability were examined with the use of MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) and NR (neutral red) methods. Induction of necrosis was assessed by propidium iodide/Hoechst staining. Progress of the cell cycle involved in cell proliferation, apoptosis, and regulation of transcription was estimated using flow cytometry analysis. Additionally, the capability of free radical scavenging was evaluated with the DPPH assay. Results. The study revealed that extracts inhibited the proliferation of cancer cells. The NR study confirmed the low cytotoxic activity of the tested extracts to normal human colon epithelial cells (CCD 841 CoTr) and human skin fibroblasts (HSF). Furthermore, a dose-dependent cytotoxicity against HT-29 cells, but not A549 cells, has been reported. The free radical scavenging activity was observed in the case of the HWE but not the HJE. Conclusions. The obtained results indicate a cancer chemopreventive potential of young barley as a safe dietary agent in colon carcinoma.

FAM83D knockdown regulates proliferation, migration and invasion of colorectal cancer through inhibiting FBXW7/Notch-1 signalling pathway.

Colorectal cancer is one of the most common malignant tumors in the digestive system, and in China its incidence is rising in recent years. The FAM83D family with sequence similarity 83, member D is associated with the occurrence and development of various cancers. However, in human colorectal cancer, the biological function of FAM83D and its molecular mechanism are still little known. In our study, we found that FAM83D mRNA expression level was markedly up-regulated in colorectal cancerous tissues when compared with that of adjacent normal colon tissues. We also found that the protein and mRNA expression levels of FAM83D are dramatically increased in human colorectal cancer cell lines, including Caco-2, RKO, DLD-1, HT-29, LoVo, SW480, and HCT116, especially in SW480 and HCT116 cells, when compared with that of human normal colon epithelial cell line (NCM460). Next, in HCT116 and SW480 cells, the biological function of FAM83D was examined. FAM83D-knockdown notably inhibited cell proliferation and colony formation. Cell apoptosis was promoted by FAM83D knockdown. In addition, FAM83D siRNA decreased cell migration and invasion. Moreover, FAM83D knockdown up-regulated the protein expression level of F-box and WD repeat domain-containing 7 (FBXW7), but diminished the Notch1 protein expression level. It also found that FBXW7 siRNA reverses the suppressive effect of FAM83D knockdown on Notch1 protein expression. Notch1 overexpression reversed the effect of FAM83D knockdown on colorectal cancer cell proliferation, cell migration and invasion. In conclusion, FAM83D knockdown promoted colorectal cancer cell apoptosis, inhibited cell proliferation, cell migration and invasion, which might be associated with inhibiting the FBXW7/Notch1 signal pathway. Our findings indicated that FAM83D is a promising molecular target for colorectal cancer treatment.

Comparative Cytotoxicity of Six Iodinated Disinfection Byproducts on Nontransformed Epithelial Human Colon Cells

Exposure to disinfection byproducts (DBPs) in chlorinated drinking water has been positively associated with increased risk of colon, bladder, and rectum cancers. Iodinated DBPs (I-DBPs) are of concern as this class exhibits enhanced cytotoxicity and genotoxicity compared to those of chlorinated and brominated equivalents in Chinese hamster ovary (CHO) cells. We tested the impact of six I-DBPs on immortalized normal human colon epithelial cells (CCD 841 CoN). Our assay showed the following rank order of cytotoxicity of the I-DBPs: iodoacetic acid (IAA) > iodoacetamide (IAcAm) > bromoiodoacetamide (BIAcAm) > chloroiodoacetamide (CIAcAm) > bromoiodoacetic acid (BIAA) ≈ diiodoacetic acid (DIAA). The enhanced cytotoxicity of IAA compared with those of other haloacetic acids agrees with studies conducted on CHO cells. IAcAm was found to be 3.5 times more cytotoxic than BIAcAm and 9.4 times more cytotoxic than CIAcAm. The cytotoxicity of both dihaloacids (i.e., BIAA and DIAA) was <1% of that of the monohaloacid...

NCAPH plays important roles in human colon cancer

Colon cancer (CC) is one of the major malignancies worldwide, whose pathogenesis is complex and requires the accumulated alteration of multiple genes and signaling pathways. Condensins are multi-protein complexes that play pivotal roles in chromosome assembly and segregation during mitosis, meiosis and even tumorigenesis. Using tissue microarrays by immunohistochemistry and hematoxylin–eosin staining, we found that non-SMC condensin I complex subunit H (NCAPH) in colon cancerous tissues was higher than that in all corresponding adjacent non-cancerous tissues. We then characterized the exact function of the NCAPH in CC. We provided evidences showing that NCAPH is highly expressed in colorectal cancer cell lines comparing with normal human colonic epithelial cells, and identified many NCAPH mutations in CC patients. We found that depletion of NCAPH inhibits CC cell proliferation, migration in vitro and xenograft tumor formation in vivo. Furthermore, NCAPH knockdown promotes cell apoptosis and cell cycle arrest at G2/M phase. Interestingly, the NCAPH high expression in tumor tissues of colon patients had a significantly better prognosis and survival rate than low-expression patients, suggesting that NCAPH high expression promotes colonic cancerous cell proliferation; on the other hand, it may also sensitize these cells responding to chemo- or radio-therapies. Collectively, these findings reveal an important role of NCAPH in CC, indicating that NCAPH could be used as a new therapeutic target in future.

Decrease of miR-622 expression suppresses migration and invasion by targeting regulation of DYRK2 in colorectal cancer cells.

BackgroundMore and more evidence indicates that microRNAs are present and involved in many tumor-related diseases. The function of microRNA-622 (miR-622) in colorectal cancer (CRC) remains controversial. Dual specificity tyrosine phosphorylation-regulated kinase 2 (DYRK2) has been reported as a tumor suppressor gene in different cancers. The detailed regulation mechanism of DYRK2 in CRC remains unclear.MethodsmiR-622 and DYRK2 expression levels were detected at tissue and cellular level respectively by using real time polymerase chain reaction (PCR), Western blot, and immunohistochemical staining. Pearson’s correlation analysis was used to evaluate the correlation between miR-622 and DYRK2. Transwell assay was applied to measure the effect of miR-622 on migration and invasion of SW1116 and SW480. We used dual luciferase reporter assay to confirm the targeted binding effect of miR-622 and DYRK2 3′-untranslated region (3′UTR). An antisense experiment was executed to further confirm the role miR-622 had played with regard to migration and invasion by targeting regulation of DYRK2 pathway in CRC cells.ResultsIn our research, we found that the expression of miR-622 was elevated in CRC tissues and cell lines compared to that of nonCRC tissues and the normal human colon epithelial cell line NCM460. Correspondingly, the expression of DYRK2 in CRC tissues and cell lines showed a contrary tendency. The different expression level of DYRK2 was closely correlated with clinicopathological characteristics of CRC patients. We demonstrated that down-regulation of miR-622 could inhibit the ability of migration and invasion of CRC cell lines SW1116 and SW480. Also, we confirmed that DYRK2 was negatively regulated by miR-622 via a specific targeted binding site within the 3′UTR. We finally verified that the migration and invasion ability of CRC cells in the conducted DYRK2 3′UTR defect plasmid transfection group were lower compared to miR-622 and cotransfection group.ConclusionThe findings of this study indicate that a decrease of miR-622 expression could suppress migration and invasion by targeting regulation of DYRK2 and miR-622/DYRK2 could be a potential molecular treating target of CRC.

Peroxiredoxin 2 is associated with colorectal cancer progression and poor survival of patients

The present study was to investigate the clinical significance of peroxiredoxin 2 (PRDX2), an oncoenzyme, in the development and progression of colorectal cancer(CRC).We found levels of PRDX2 mRNA and protein were higher in CRC cell lines than in normal human colonic epithelial cells. PRDX2 expression was significantly up-regulated in CRC lesions compared with that in the adjacent noncancerous tissues. CRC tissues from 148 of 226 (65.5%) patients revealed high level of PRDX2 protein expression in contrast to only 13 of 226 (5.8%) PRDX2 strong staining cases in the adjacent noncancerous tissues. Increased expression of PRDX2 protein was significantly associated with poor tumor differentiation (p = 0.001), advanced local invasion (p = 0.046), increased lymph node metastasis (p = 0.008), and advanced TNM stage (p = 0.020). Patients with higher PRDX2 expression had a significantly shorter disease-free survival and worse disease-specific survival than those with low expression. Importantly, PRDX2 up-regulation was an independent prognostic indicator for stage I–III, early stage (stage I-II) and advanced stage (stage III) patients. In conclusion, our findings suggest PRDX2 up-regulation correlates with tumor progression and could serve as a useful marker for the prognosis of CRC.

Bile acid: a potential inducer of colon cancer stem cells.

BackgroundAlthough the unconjugated secondary bile acids, specifically deoxycholic acid (DCA) and lithocholic acid (LCA), are considered to be risk factors for colorectal cancer, the precise mechanism(s) by which they regulate carcinogenesis is poorly understood. We hypothesize that the cytotoxic bile acids may promote stemness in colonic epithelial cells leading to generation of cancer stem cells (CSCs) that play a role in the development and progression of colon cancer.MethodsNormal human colonic epithelial cells (HCoEpiC) were used to study bile acid DCA/LCA-mediated induction of CSCs. The expression of CSC markers was measured by real-time qPCR. Flow cytometry was used to isolate CSCs. T-cell factor/lymphoid-enhancing factor (TCF/LEF) luciferase assay was employed to examine the transcriptional activity of β-catenin. Downregulation of muscarinic 3 receptor (M3R) was achieved through transfection of corresponding siRNA.ResultsWe found DCA/LCA to induce CSCs in normal human colonic epithelial cells, as evidenced by the increased proportion of CSCs, elevated levels of several CSC markers, as well as a number of epithelial–mesenchymal transition markers together with increased colonosphere formation, drug exclusion, ABCB1 and ABCG2 expression, and induction of M3R, p-EGFR, matrix metallopeptidases, and c-Myc. Inhibition of M3R signaling greatly suppressed DCA/LCA induction of the CSC marker ALDHA1 and also c-Myc mRNA expression as well as transcriptional activation of TCF/LEF.ConclusionsOur results suggest that bile acids, specifically DCA and LCA, induce cancer stemness in colonic epithelial cells by modulating M3R and Wnt/β-catenin signaling and thus could be considered promoters of colon cancer.