Colon Cancer Cell Line SW480 Research Articles

Exosomes derived from colon cancer cells and plasma of colon cancer patients promote migration of SW480 cells through Akt/mTOR pathway

ObjectiveTo investigate the effect of exosomes derived from colon cancer (CC) cells and plasma of CC patients on migration of SW480 cells. MethodsThe exosomes derived from culture medium of human colon epithelial cell line NCM460 and CC cell line SW620 were isolated by ultracentrifugation. The exosomes derived from plasma of CC patients and healthy controls were isolated by size exclusion chromatography (SEC). The particle size and morphology of exosomes were identified by nanoparticle tracking analysis (NTA) and transmission electron microscopy (TEM) respectively, and exosomal markers were detected by Western blotting. The uptake of fluorescent DiI labeled exosomes by SW480 cells was observed by confocal microscopy. Transwell assay was used to detect the effect of exosomes on the migration of SW480 cells. The expression level of associated proteins in signaling pathway were analyzed by Western blotting. Rapamycin, an inhibitor of mTOR, was used to study the role of mTOR signaling pathway on exosomes mediated migration of SW480 cells. ResultsThe results of NTA and TEM showed that the particle size of the isolated exosomes was about 120 nm, which were small vesicles with membrane structure. The expressions of exosomal markers Alix, TSG101 and CD63 could be detected. The exosomes were evidenced by a red fluorescent signal inside the cytoplasm of SW480 recipient cells, and could promote the migration of SW480 cells, which is associated with Akt/mTOR signaling pathway. Compared with the control group, plasma exosomes derived from CC patients could significantly promote the migration of SW480 cells. Inhibition the activity of mTOR signaling could attenuate the migration of SW480 cells. ConclusionsExosomes derived from CC cells and plasma of CC patients could promote the migration of SW480 cells, which is associated with Akt/mTOR signaling pathway.

Evaluating the effect of dermaseptin S4 and its derivatives on multidrug-resistant bacterial strains and on the colon cancer cell line SW620.

Dermaseptins are peptides found in the skin secretions of Phyllomedusinae frogs. These peptides exert a lytic action on various microorganisms and have no considerable hemolytic effect except dermaseptin S4 (DS4) which exhibits a powerful cytotoxic effect. Therefore, we synthesized several analogs of DS4 in an attempt to find molecules with a weak hemolytic effect and significant bioactivities. In this study, we performed the synthesis of truncated peptides by introducing C-terminal and N-terminal amino acid deletions of the native sequence. All peptide analogs, in comparison with parental peptide, were tested firstly on human red blood cells to work out their cytotoxicity, secondly on the multidrug-resistant bacteria by trying to find MICs, and finally on colon cancer tumor cell line SW620 using the MTT test so as to investigate the anti-proliferative effect. Our results showed that, on the one hand, the N terminus of the native peptide was necessary for the antibacterial activity and the anti-proliferative effect of the peptide. On the other hand, the hemolytic activity was more notable in the sequences broken down on the C-terminal side.

RGD4C peptide mediates anti-p21Ras scFv entry into tumor cells and produces an inhibitory effect on the human colon cancer cell line SW480

BackgroundWe prepared an anti-p21Ras scFv which could specifically bind with mutant and wild-type p21Ras. However, it cannot penetrate the cell membrane, which prevents it from binding to p21Ras in the cytoplasm. Here, the RGD4C peptide was used to mediate the scFv penetration into tumor cells and produce antitumor effects.MethodsRGD4C-EGFP and RGD4C-p21Ras-scFv recombinant expression plasmids were constructed to express fusion proteins in E. coli, then the fusion proteins were purified with HisPur Ni-NTA. RGD4C-EGFP was used as reporter to test the factors affecting RGD4C penetration into tumor cell. The immunoreactivity of RGD4C-p21Ras-scFv toward p21Ras was identified by ELISA and western blotting. The ability of RGD4C-p21Ras-scFv to penetrate SW480 cells and colocalization with Ras protein was detected by immunocytochemistry and immunofluorescence. The antitumor activity of the RGD4C-p21Ras-scFv was assessed with the MTT, TUNEL, colony formation and cell migration assays. Chloroquine (CQ) was used an endosomal escape enhancing agent to enhance endosomal escape of RGD4C-scFv.ResultsRGD4C-p21Ras-scFv fusion protein were successfully expressed and purified. We found that the RGD4C fusion protein could penetrate into tumor cells, but the tumor cell entry of was time and concentration dependent. Endocytosis inhibitors and a low temperature inhibited RGD4C fusion protein endocytosis into cells. The change of the cell membrane potential did not affect penetrability. RGD4C-p21Ras-scFv could penetrate SW480 cells, effectively inhibit the growth, proliferation and migration of SW480 cells and promote this cells apoptosis. In addition, chloroquine (CQ) could increase endosomal escape and improve antitumor activity of RGD4C-scFv in SW480 cells.ConclusionThe RGD4C peptide can mediate anti-p21Ras scFv entry into SW480 cells and produce an inhibitory effect, which indicates that RGD4C-p21Ras-scFv may be a potential therapeutic antibody for the treatment of ras-driven cancers.

Cytotoxicity and Apoptosis Effects of Curcumin Analogue (2E,6E)-2,6-Bis(2,3-Dimethoxybenzylidine) Cyclohexanone (DMCH) on Human Colon Cancer Cells HT29 and SW620 In Vitro.

Colorectal cancer (CRC) is the third most common type of cancer worldwide and a leading cause of cancer death. According to the Malaysian National Cancer Registry Report 2012–2016, colorectal cancer was the second most common cancer in Malaysia after breast cancer. Recent treatments for colon cancer cases have caused side effects and recurrence in patients. One of the alternative ways to fight cancer is by using natural products. Curcumin is a compound of the rhizomes of Curcuma longa that possesses a broad range of pharmacological activities. Curcumin has been studied for decades but due to its low bioavailability, its usage as a therapeutic agent has been compromised. This has led to the development of a chemically synthesized curcuminoid analogue, (2E,6E)-2,6-bis(2,3-dimethoxybenzylidine) cyclohexanone (DMCH), to overcome the drawbacks. This study aims to examine the potential of DMCH for cytotoxicity, apoptosis induction, and activation of apoptosis-related proteins on the colon cancer cell lines HT29 and SW620. The cytotoxic activity of DMCH was evaluated using the [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] (MTT) cell viability assay on both of the cell lines, HT29 and SW620. To determine the mode of cell death, an acridine orange/propidium iodide (AO/PI) assay was conducted, followed by Annexin V/FITC, cell cycle analysis, and JC-1 assay using a flow cytometer. A proteome profiler angiogenesis assay was conducted to determine the protein expression. The inhibitory concentration (IC50) of DMCH in SW620 and HT29 was 7.50 ± 1.19 and 9.80 ± 0.55 µg/mL, respectively. The treated cells displayed morphological features characteristic of apoptosis. The flow cytometry analysis confirmed that DMCH induced apoptosis as shown by an increase in the sub-G0/G1 population and an increase in the early apoptosis and late apoptosis populations compared with untreated cells. A higher number of apoptotic cells were observed on treated SW620 cells as compared to HT29 cells. Human apoptosis proteome profiler analysis revealed upregulation of Bax and Bad proteins and downregulation of Livin proteins in both the HT29 and SW620 cell lines. Collectively, DMCH induced cell death via apoptosis, and the effect was more pronounced on SW620 metastatic colon cancer cells, suggesting its potential effects as an antimetastatic agent targeting colon cancer cells.

Inhibition of BAMBI reduces the viability and motility of colon cancer via activating TGF-β/Smad pathway in vitro and in vivo.

[This retracts the article DOI: 10.3892/ol.2017.6811.].

Changes of Kinesin Family Member 18 Pathway in Epithelial Mesenchymal Transformation and Effect of Butyrate on Migration and Invasion of Colon Cancer Cells

Colon cancer is a common digestive system disease with an increasing incidence. Severe migration and invasion aggravates the deterioration of colon cancer patients. Previous studies have found that epithelial mesenchymal transition (EMT) is closely associated with early transference of colon carcinoma and abnormal changes occur in KIF18 signaling pathway. Butyrate protects colonic mucosa with a considerable effect on the colon. This study predicts that butyrate may reverse EMT process of colon cancer cells through KIF18 signaling pathway, thereby inhibiting cell migration and invasion. In this experiment, EMT model of colon cancer was used to investigate migration and invasion. Human colon cancer cell line SW1116 was cultured and assigned into control group (0 mmol/L butyrate), low concentration group (2 mmol/L), medium concentration group (4 mmol/L), and high concentration group (10 mmol/L). After 72 hours, cell migration and invasion was analyzed by Transwell assays. E-cadherin, Vimentin, and KIF18 level was detected by Western blot and quantitative real-time PCR. After treatment, cell migration and invasion was significantly inhibited compared to control dose-dependently. In addition, Vimentin and KIF18 mRNA level was significantly lower and E-cadherin mRNA was higher in treatment groups than control group in a dose-dependent manner (P < 0.05). Consistently, the profile of protein level of these molecules was similar to mRNA expression profile. Electron microscope showed that after treatment with butyrate, the surface protuberances of colon cancer cells were abnormally increased, especially the vesicular protuberances, which were the microvilli of intestinal mucosal epithelium. In conclusion, KIF18 is crucial in EMT of colon cancer cells. Butyrate may elevate E-cadherin and suppress Vimentin and KIF18 by activating KIF18 signaling, thus inhibiting invasion and migration.

Application of decoy oligodeoxynucleotides strategy for inhibition of cell growth and reduction of metastatic properties in nonresistant and erlotinib-resistant SW480 cell line.

Signal transducer and activator of transcription 3 (STAT3) is a critical regulator for angiogenesis, cell cycle progression, apoptosis, and drug resistance. Resistance toward EGF receptor (EGFR) inhibitors is a significant clinical concern for metastatic colon cancer patients. The present study aimed to evaluate the blocking influences of STAT3 decoy oligodeoxynucleotides (ODNs) on the STAT3 survival signaling pathway in nonresistant and erlotinib-resistant SW480 colon cancer cells. First, STAT3 decoy and scramble ODNs were designed according to STAT3 elements in the promoter region of MYCT1 gene and tested for the interaction of STAT3 protein with designed ODNs via in silico molecular docking study. Then, the efficiency of transfection and subcellular localization of ODNs were assessed using flow cytometry and fluorescence microscopy, respectively. Cell viability, cell cycle, and apoptosis tests, scratch and colony formation assays, and real-time PCR were also used to study the cancerous properties of cells. A considerable decrease in proliferation of colon cancer cells was observed with blockade of STAT3 signaling due to cell cycle arrest and induced apoptosis via downregulation of cyclin D1 and Bcl-XL, respectively. Furthermore, upon transfecting STAT3 decoy ODNs, colony formation potential and migration activity in both SW480 colon cancer cell lines were decreased compared to the control groups. From this study, it could be concluded that STAT3 is critical for cell growth inhibition and metastatic properties reduction of resistant SW480 colon cancer cells; therefore, STAT3 decoy ODNs could be considered as potential therapeutics along with current remedies for treating drug-resistant colon cancer.

L-fucose ameliorates the carcinogenic properties of Fusobacterium nucleatum in colorectal cancer.

Fusobacterium nucleatum (Fn) is considered a promoting factor in colorectal cancer (CRC); however, only a few studies have investigated therapies against Fn. L-fucose is a natural monosaccharide that has prebiotic potential. The present study aimed to investigate the effect of L-fucose on the carcinogenic properties of Fn. The HCT116 and SW480 colon cancer cell lines were treated with Fn and Fn+L-fucose (Fnf), respectively. The Cell Counting Kit-8 (CCK-8), colony formation, Transwell migration and invasion and wound healing assays were performed to assess the proliferative, migratory and invasive abilities of the cells, respectively. Western blot was performed to detect the protein levels of jak/stat3 pathway components and EMT. The results of the CCK-8, colony formation, Transwell and wound healing assays demonstrated that treatment with Fn significantly enhanced the proliferative, migratory and invasive abilities of HCT116 and SW480 colon cancer cells. Notably, these effects were significantly reversed following addition of L-fucose. Furthermore, L-fucose inhibited the carcinogenic properties of Fn to activate the stat3 pathway and epithelial-to-mesenchymal transition. Taken together, the results of the present study suggest that L-fucose ameliorates the carcinogenic properties of Fn in vitro, and thus may serve as a novel therapeutic target for flora-related colon cancer.

Epibrassinolide activates AKT to trigger autophagy with polyamine metabolism in SW480 and DLD-1 colon cancer cell lines.

Epibrassinolide (EBR), a plant-derived polyhydroxylated derivative of 5α-cholestane, structurally shows similarities to animal steroid hormones. According to the present study, EBR treatment triggered a significant stress response via activating ER stress, autophagy, and apoptosis in cancer cells. EBR could also increase Akt phosphorylation in vitro. While the activation of Akt resulted in cellular metabolic activation in normal cells to proceed with cell survival, a rapid stress response was induced in cancer cells to reduce survival. Therefore, Akt as a mediator of cellular survival and death decision pathways is a crucial target in cancer cells. In this study, we determined that EBR induces stress responses through activating Akt, which reduced the mTOR complex I (mTORC1) activation in SW480 and DLD-1 colon cancer cells. As a consequence, EBR triggered macroautophagy and led to lipidation of LC3 most efficiently in SW480 cells. The cotreatment of spermidine (Spd) with EBR increased lipidation of LC3 synergistically in both cell lines. We also found that EBR promoted polyamine catabolism in SW480 cells. The retention of polyamine biosynthesis was remarkable following EBR treatment. We suggested that EBR-mediated Akt activation might determine the downstream cellular stress responses to induce autophagy related to polyamines.

Overexpression of protein regulator of cytokinesis 1 facilitates tumor growth and indicates unfavorable prognosis of patients with colon cancer

BackgroundProtein regulator of cytokinesis 1 (PRC1) has been reported to play important role in the pathogenesis of various cancers. However, its role in colon cancer has not been studied. Here, we aimed to investigate the biological functions and potential mechanism of PRC1 in colon cancer.MethodsThe expression level of PRC1 in colon cancer tissues and cell lines was detected by quantitative real-time polymerase chain reaction (qRT-PCR), Western blotting, and immunohistochemical (IHC) staining of a tissue microarray (TMA). Furthermore, colon cancer cell lines HCT116 and SW480 were treated with short hairpin RNAs against PRC1. The biological function of PRC1 was determined by MTT proliferation, colony formation assay, cell cycle, and apoptosis assays. Then, an in vivo tumor formation assay was conducted to explore the effects of PRC1 on tumor growth.ResultsThe mRNA and protein expression levels of PRC1 were highly expressed in colon cancer tissues and cell lines. PRC1 expression was associated with clinicopathological characteristics and overall survival of patients with colon cancer. Knockdown of PRC1 could decrease proliferation and colony forming ability of colon cancer cells, as well as arrested more cells at G2/M phase and promoted cell apoptosis. In cancer cells, the expression pattern of protein regulators included in cell cycle and apoptosis progress were reverted by PRC1 down-regulation. Additionally, PRC1 down-regulation could suppress colon tumor growth and differentiation.ConclusionsWe confirmed that PRC1 was overexpressed in colon cancer and was associated with poor prognosis of colon cancer patients. PRC1 down-regulation could arrest cell cycle at G2/M stage, inhibit proliferation, and elicit apoptosis. These findings showed the potential of PRC1 to be used for therapeutic approaches in colon cancer.

Non-thermal effects of radiofrequency electromagnetic fields

We explored the non-thermal effects of radiofrequency (RF) electromagnetic fields and established a theoretical framework to elucidate their electrophysiological mechanisms. In experiments, we used a preclinical treatment device to treat the human colon cancer cell lines HT-29 and SW480 with either water bath heating (WB-HT) or 13.56 MHz RF hyperthermia (RF-HT) at 42 °C for 60 min and analyzed the proliferation and clonogenicity. We elaborated an electrical model for cell membranes and ion channels and estimated the resulting ion fluxes. The results showed that, for both cell lines, using RF-HT significantly reduced proliferation and clonogenicity compared to WB-HT. According to our model, the RF electric field component was rectified and smoothed in the direction of the channel, which resulted in a DC voltage of ~ 1 µV. This may induce ion fluxes that can potentially cause relevant disequilibrium of most ions. Therefore, RF-HT creates additional non-thermal effects in association with significant ion fluxes. Increasing the understanding of these effects can help improve cancer therapy.

Enhanced drug retention, sustained release, and anti-cancer potential of curcumin and indole-curcumin analog-loaded polysorbate 80-stabilizied PLGA nanoparticles in colon cancer cell line SW480.

The major therapeutic limitation of curcumin and indole-incorporated curcumin analog is its low bioavailability. We hypothesized that nano-encapsulation of indole-incorporated curcumin analog and curcumin as a biodegradable polymeric nanoparticle may enhance its bioavailability with extended drug retention time. Indole-incorporated curcumin analog and curcumin loaded PLGA nanoparticles were synthesized by solvent evaporation technique. Physicochemical characterizations and anti-cancer potential of the nanoparticles were evaluated in human colon cancer cell line SW480. The synthesized NPs had a size range of 50-150nm diameter. The nano-formulation preserved the drug from degradation in wide ranges of pH environments. The nanoparticles treatment against SW480 cancer cell line triggered nuclear fragmentation, cell cycle blockade, inhibition of apoptosis and metastatic biomarkers. These drug-loaded nanoparticles may be potent nano-formulations against colon cancer because of its ability to tolerate extreme pH environments, thus having potential of oral drug-delivery.

Identification of biomarkers associated with extracellular vesicles based on an integrative pan-cancer bioinformatics analysis.

Extracellular vesicle (EV) has received increasing attention over the last decade. However, biomarkers and mechanisms underlying remain largely limited. Three microarray profiles, GSE78718 (K562 leukemia cell line), GSE45301 (U87-MG glioblastoma cell line), and GSE9589 (SW480 colon cancer cell line), were analyzed for the overlapped differentially expressed genes (DEGs). SurvExpress was used for the prognostic analysis of hub genes signature. Predicted transcription factors networks were built by NetworkAnalysis. Characterization between hub genes and immune cells was analyzed by the tumor immune estimation resources (TIMER) and single-sample gene set enrichment analysis (ssGSEA). The most significantly enriched pathway was lysosome. Hub genes included lysosomal-associated membrane protein 1 (LAMP1), heat shock protein family A (Hsp70) member 5 (HSPA5), lysosomal-associated membrane protein 2 (LAMP2), integrin subunit alpha V (ITGAV), and transmembrane protein 30A (TMEM30A). Significant prognostic values of hub genes signature were identified in glioblastoma (P-value = 0.006), but not colon cancer. In colon cancer, ITGAV displayed remarkably high correlation with tumor immune infiltrating cells. In glioblastoma, the highest correlation was found between HSPA5 and dendritic cell. Moreover, distinct association of immune cells between cell and EV were identified via ssGSEA. This study identified biomarkers in EV with potential immunological insights and clinical values.

Itraconazole inhibits the Hedgehog signaling pathway thereby inducing autophagy-mediated apoptosis of colon cancer cells

Itraconazole is as an antifungal medication used to treat systemic fungal infections. Recently, it has been reported to be effective in suppressing tumor growth by inhibiting the Hedgehog signaling pathway and angiogenesis. In the present study, we investigated whether itraconazole induces autophagy-mediated cell death of colon cancer cells through the Hedgehog signaling pathway. Cell apoptosis and cell cycle distribution of the colon cancer cell lines SW-480 and HCT-116 were detected by flow cytometry and terminal TUNEL assay. Autophagy and signal proteins were detected by western blotting and cell proliferation-associated antigen Ki-67 was measured using immunohistochemistry. The images of autophagy flux and formation of autophagosomes were observed by laser scanning confocal and/or transmission electron microscopy. Colon cancer cell xenograft mouse models were also established. Itraconazole treatment inhibited cell proliferation via G1 cell cycle arrest as well as autophagy-mediated apoptosis of SW-480 and HCT-116 colon cancer cells. In addition, the Hedgehog pathway was found to be involved in activation of itraconazole-mediated autophagy. After using the Hedgehog agonist recombinant human Sonic Hedgehog (rhshh), itraconazole could counteract the activation of rhshh. Moreover, treatment with itraconazole produced significant cancer inhibition in HCT-116-bearing mice. Thus, itraconazole may be a potential and effective therapy for the treatment of colon cancer.

RETRACTED ARTICLE: A novel small-molecule inhibitor suppresses colon cancer metastasis through inhibition of metastasis-associated in colon cancer-1 transcription.

We previously reported that the metastasis-associated in colon cancer-1 (MACC-D1) is overexpressed in colon cancer. However, so far, only a few effective MACC-1 inhibitors have been identified. To discover novel transcriptional inhibitors of MACC-1, we performed a luciferase reporter-based high-throughput screening (HTS). This strategy discovered rottlerin as an inhibitor of MACC-1 transcription was able to reduce cell motility in colon cancer in time- and dose-dependent manners. Wound healing assay indicated that 48-h treatment with 2.5 μM rottlerin impairs wound closure compared to the controls. Electrophoretic mobility shift assays showed that rottlerin inhibits the binding of the transcriptional activators Sp1 and c-Jun with human MACC-1 promoter. A combination of Western blot assays and expression analyses by qRT-PCR ruled out that rottlerin may act indirectly through inhibition of c-Jun or Sp1 expression to decrease MACC-1 expression in human colon cancer cell lines SW620. Rather, these results support that rottlerin restricts the binding of MACC-1 promoter directly by c-Jun and Sp1 in colon cancer cells. Thus, as a small-molecule inhibitor of MACC-1, rottlerin may benefit colon cancer patients by suppressing MACC-1-dependent tumor growth and metastasis.

The Role and Mechanism of S1PR5 in Colon Cancer.

PurposeTo investigate the role and mechanism of S1PR5 in colon cancer.Materials and MethodsLentiviral infection and drug screening helped to establish colon cancer cell lines with stable overexpression and knockdown of S1PR5. Effects of S1PR5 expression on cell growth, proliferation, migration, and invasion were analyzed using a subcutaneous xenograft model in nude mice. Western blot (WB) was used to detect the effects of S1PR5 expression on p-AKT, STAT3, NF-κB, and p-JNK. The distribution of p65 was evaluated in nuclear and cytoplasmic fractions using WB. CCK-8, Transwell migration, and Transwell invasion assays analyzed cell growth, proliferation, migration, and invasion. qRT-PCR analysis revealed that S1PR5 expression was associated with altered expression levels of NF-κB downstream target genes, such as IL-6, TNF-α, and indoleamine 2, 3-dioxygenase 1 (IDO1).ResultsqRT-PCR and WB analysis showed that the S1PR5 level in colon cancer cell lines—SW480, SW620, HCT116, and LoVo—was significantly higher than in NCM460, a healthy colonic epithelial cell line. SW620 and SW480, with high and low expression of S1PR5, respectively, were selected as model cell lines. S1PR5 knockdown in SW620 caused the growth rate, proliferation, migration, invasion, and subcutaneous tumor formation rate to decrease in mice, whereas S1PR5 overexpression in SW480 caused all of these parameters to increase. WB analysis showed an increase in phospho-p65 and its nuclear translocation. S1PR5 knockdown caused a decrease in phospho-p65 levels and its nuclear import, thereby inhibiting its activity. In S1PR5 knockdown and overexpressing cells, p65 was overexpressed and knocked down, respectively. qRT-PCR and WB showed that S1PR5 over-expression up-regulates IDO1, and S1PR5 knockdown inhibits IDO1. CCK-8 and Transwell assays showed that p65 and IDO1 overexpression antagonizes the antitumor effect of S1PR5 knockdown, and that p65 and IDO1 knockdown antagonizes the tumorigenic effect of S1PR5 overexpression.ConclusionS1PR5 overexpression promotes the growth, migration, and invasion of cancer by activating the NF-κB/IDO1 signaling pathway.

Filago germanica (L.) Huds. bioactive constituents: Secondary metabolites fingerprinting and in vitro biological assays

Plants from Asteraceae have interesting medicinal potential in traditional medicines and have been studied for different pharmacological effects. This study attempts to describe the phytochemical and biological profiles of dichloromethane (DCM) and methanol (MeOH) extracts of aerial parts and roots of Filago germanica (L.) Huds. Phytochemical composition was assessed by high performance liquid chromatography photodiode array (HPLC-PDA) polyphenolic quantification, and ultra-high performance liquid chromatography mass spectrometry (UHPLC-MS) secondary metabolites analysis. Biological potential were determined via antioxidant (2, 2′-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid -ABTS, cupric reducing antioxidant capacity -CUPRAC and metal chelation), enzyme inhibition (α-amylase, tyrosinase and lipoxygenase -LOX) and cytotoxic (SW-480 colon cancer cell line) assays. The HPLC-PDA quantification identified 11 phenolic phytochemicals, amongst epicatechin (1.81 μg/g extract) and syringic acid (2.23 μg/g extract) were present in higher amounts. Similarly, UHPLC-MS analysis of both DCM extracts showed the tentative presence of several flavonoid, alkaloid and terpenoid derivatives as major components. The aerial-MeOH extract was found to possess highest ABTS and CUPRAC antioxidant potential with values of 119.11 and 170.83 mg Trolox equivalent (TE)/g extract, respectively. Root-DCM extract was the most active in the metal chelation assay (22.86 mg ethylenediaminetetraacetic acid equivalent (EDTAE)/g extract). The root-DCM extract was most active against tyrosinase (30.51 mg kojic acid equivalent (KAE)/g extract) and α-amylase (0.30 acarbose equivalent (ACAE)/g extract), while aerial-MeOH extract showed maximum LOX inhibition. Moreover, the aerial parts and root DCM extracts were most active against colon cancer cell line with half maximal inhibitory concentration (IC50) of 26.95 and 41.62 μg/mL, respectively. Based on the present findings, F. germanica can be viewed as a promising bio-resource for bioprospecting of novel pharmaceuticals and biomedicine industrial products.

Inhibition of colorectal tumor formation using a new miR‐210 therapeutic inhibitor that targets the Lnc RNA Xist and Nme1

In this report we investigate the role of microRNA‐210 (miR‐210) in the etiology of colorectal cancer (CRC) using both cell‐based and xenograft mouse models. We show that miR‐210 expression is higher in the colon cancer cell line SW620 versus normal colon epithelium and has been associated with decreased cancer survival rates. Stable expression of our new miR‐210 inhibitor changes the morphology, proliferation and gene expression of SW620 cells and tumor growth. RNAseq identified potential targets of miR‐210 and qPCR has verified several of these targets. We find that inhibition of miR‐210 can act as a tumor suppressor and reduce tumor formation of SW620 colon cancer cells in xenograft mouse models. Direct comparison with capecitabine treatments demonstrates the effectiveness of the miR‐210 inhibitor treatment. The direct administration of the miR‐210 inhibitor (MiRIS) to existing tumors can inhibit tumor growth in both NSG and Foxnu/j mouse models. The inhibition of miR‐210 can cause alterations in hypoxia responses and upregulation of several tumor suppressor genes, including Nme1. We demonstrate that the long non‐coding transcript XIST is regulated by miR‐210 and XIST expression is decreased in colon tumors. We show that XIST expression correlates with the expression of the tumor suppressor gene Nme1. Furthermore, the inhibition of miR‐210 effects epigenetic marks of the NME1 loci, with increases in H3K4me3 and H3K27ac near the transcription start site (TSS). We show that our new miR‐210 inhibitor can be used as an effective colorectal cancer therapeutic tool.

Effects of propofol on invasion and migration of colon cancer cells and JAK2/STAT3 signaling pathway.

To investigate the effect of propofol on invasion and migration of human colon cancer cell line SW480 and the JAK2/STAT3 pathway. Human colon cancer cell line SW480 was divided into a control group, 3 propofol treatment groups (with 2, 4 and 8 μg/mL propofol treatment, respectively), and a propofol+colivelin group. Lactate dehydrogenase (LDH) activity and cell proliferation was evaluated by detection kit and 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide (MTT) method, respectively. Cell migration and invasion were detected by Transwell assay. Western blotting was performed to detect the protein expression levels of JAK2, p-JAK 2, STAT3 and p-STAT3 in colon cancer cell line SW480. Compared with the control group, the LDH activity was significantly increased in the propofol treatment groups (P<0.05). The cell proliferation, migration, invasion as well as p-JAK2 and p-STAT3 in colon cancer cell line SW480 in the propofol treatment groups were significantly decreased (allP<0.05). Compared with the propofol treatment groups, the p-STAT3 level, proliferation, migration and invasion in the colon cancer cell line SW480 were significantly increased in the propofol+colivelin group (allP<0.05). Propofol can inhibit the proliferation, migration and invasion of human colon cancer cells by inhibiting JAK2/STAT3 signaling pathway.

New Hybrids Derived from the Natural Compound (-)-β-Pinene and Amides or Acylthioureas as Antitumor Agents

Background:Plant-derived natural compounds have a unique molecular structure and rich biological activity, hence, they are treated as important raw materials for the development of drugs.Methods:A natural compound (-)-β-pinene was used as a raw material, and twenty-six novel derivatives with amide or acylthiourea groups were synthesized based on the molecular hybridization method. In vitro antitumor activity of these derivatives on human breast cancer cell line MCF7 and human colon cancer cell line SW1116 were tested by MTT method. The effects of the synthesized derivatives on the morphology of MCF7 and SW1116 were observed by fluorescent inverted microscope.Results:The preliminary structure-activity relationship analysis demonstrates that the position and species of substituents on the aromatic ring of derivatives have an effect on the antitumor activity of derivatives. Observation of the cell morphology reveals that derivatives with antitumor activity can lead to rounding of the cell morphology, a decrease in cell volume and cell density, and ultimately inhibition of the proliferation of MCF7 and SW1116 cells. The antitumor activity evaluation results show that among these derivatives, compounds 5c, 5e, 5h, 7c, 7b and 7e exhibit good antitumor activity against MCF7, and compounds 5c, 5e, 5h and 7j exert moderate antitumor activity against SW1116.Conclusion:This study hopes to promote the high value-added utilization of natural compounds β-pinene and the development of novel antitumor drugs.