SW480 Colon Cancer Cells Research Articles

Effect of the chemoprotectant tempol on anti-tumor activity of cisplatin

To investigate the effect of the chemoprotectant tempol on the anti-tumor activity of cisplatin (DDP). The cellular toxicity of tempol in human colon cancer SW480 cells and mouse colon cancer CT26 cells were evaluated using MTT and cell counting kit-8 assays. CalcuSyn software analysis was used to determine the interaction between tempol and DDP in inhibition of the cell viability. A subcutaneous homograft mouse model of colon cancer was established. The mice were randomly divided into control group, tempol group, cisplatin group and tempol + DDP treatment group with intraperitoneal injections of the indicated agents. The tumor size, body weight and lifespan of the mice were measured, and HE staining was used to analyze the cytotoxic effect of the agents on the kidney and liver. Immunohistochemistry and Western blotting were performed to detect the expression of Bax and Bcl2 in the tumor tissue, and TUNEL staining was used to analyze the tumor cell apoptosis. The level of reactive oxygen species (ROS) in the tumor tissue was determined using flow cytometry. Tempol showed inhibitory effects on the viability of SW480 and CT26 cells. CalcuSyn software analysis showed that tempol had a synergistic anti-tumor effect with DDP (CI < 1). In the homograft mouse model, tempol treatment alone did not produce obvious anti-tumor effect. HE staining showed that the combined use of tempol and DDP alleviated DDP-induced fibrogenesis in the kidneys, but tempol also reduced the anti-tumor activity of DDP. Compared with the mice treated with DDP alone, the mice treated with both tempol and DDP had a significantly larger tumor size (P < 0.01) and a shorter lifespan (P < 0.05). Tempol significantly reversed DDP-induced expression of Bax and Bcl2 in the tumor tissue and tumor cell apoptosis (P < 0.001), and obviously reduced the elevation of ROS level in the tumor tissue induced by DDP treatment (P < 0.05). Tempol can attenuate the anti-tumor effect of DDP while reducing the side effects of DDP. Caution must be taken and the risks and benefits should be carefully weighed when considering the use of tempol as an anti-oxidant to reduce the toxicities of DDP.

Gambogic acid suppresses colon cancer cell activity in vitro.

The aim of the present study was to elucidate the underlying mechanism of antitumor activity of gambogic acid (GA) in colon cancer. Human colon cancer SW620 cells were divided into five treatment groups, including no-treatment control (NC), low dose GA (10 µg/ml), medium dose GA (50 µg/ml), high dose GA (100 µg/ml) and 5-fluorouracil (10 µg/ml). Differences in cell proliferation, apoptosis and cell cycle, invasion, and migration were measured between groups using MTT, flow cytometry, transwell and wound-healing assays, respectively. Western blotting was used to analyze relative protein expression levels of phosphoinositide 3-kinase (PI3K), protein kinase B (AKT), P21, and matrix metalloprotease (MMP)-2 and −9 between groups. Compared with the NC group, GA (low, middle and high) inhibited SW620 cell proliferation, invasion and migration (all P<0.05). Furthermore, there were significant differences in proliferation, invasion and migration between groups administered with different doses of GA (all P<0.05). Compared with the NC group, the expression levels of PI3K, AKT, phosphorylated-AKT, P21 and MMP-2 and −9 were significantly altered in a dose dependent manner following treatment with GA (all P<0.05). The results of the current study indicated that GA suppressed proliferation and dispersion of human colon cancer cells in a dose-dependent manner, possibly through a PI3K/AKT/P21/MMP-2/9-dependent pathway.

Sulforaphane enhances apoptosis induced by Lactobacillus pentosus strain S-PT84 via the TNFα pathway in human colon cancer cells.

Sulforaphane and Lactobacilli induce apoptosis in several cancer cells. Sulforaphane, a dietary isothiocyanate, is an attractive agent due to its potent anticancer effects. Sulforaphane suppresses the proliferation of various cancer cells in vitro and in vivo. The present study investigated the effect of sulforaphane and a co-culture with Lactobacillus-treated peripheral blood mononuclear cells (PBMCs) in human colon cancer cells. The combination markedly induced apoptosis in human colon cancer HCT116 and SW480 cells. A pan-caspase inhibitor markedly inhibited apoptosis, and a tumor necrosis factor (TNF) receptor/Fc chimera partially inhibited apoptosis in both cells. The amount of TNFα secretion in the culture supernatant was significantly increased by co-culture with Lactobacillus-treated normal human PBMCs. On the other hand, the expression of cellular inhibitor of apoptosis-2 (cIAP-2), an anti-apoptotic protein, was increased by co-culture with Lactobacillus-treated PBMCs in colon cancer cells, but sulforaphane treatment significantly suppressed the induction of cIAP-2. The present results revealed that sulforaphane enhances apoptosis in human colon cancer cells under co-culture with Lactobacillus-treated PBMCs via the TNFα signaling pathway.

Monitoring of dynamic ATP level changes by oligomycin-modulated ATP synthase inhibition in SW480 cancer cells using fluorescent \u201cOn-Off\u201d switching DNA aptamer

Adenosine triphosphate (ATP) is the main energy source in cells and an important biomolecule participating in cellular reactions in living organisms. Since the ATP level changes dynamically reflecting the development of a debilitating disease or carcinogenesis, we have focused in this work on monitoring of the oligomycin (OMC)-modulated ATP synthase inhibition using a fluorescent-switching DNA aptamer designed for the detection of ATP (Apt(ATP)), as the model for studies of dynamic ATP level variation. The behavior of the ATP aptamer has been characterized using fluorescence spectroscopy. The Intramolecular fluorescence resonance energy transfer (iFRET) operates in the proposed aptamer from the FAM dye moiety to guanines of the aptamer G-quadruplex when the target ATP is present and binds to the aptamer changing its conformation. The iFRET process enables the detection of ATP down to the limit of detection, LOD = 17 μM, without resorting to any extra chemi-amplification schemes. The selectivity coefficients for relevant interferent triphosphates (UTP, GTP, and CTP) are low for the same concentration as that of ATP. We have demonstrated an efficient transfection of intact cells and OMC-treated SW480 colon cancer cells with Apt(ATP), using microscopic imaging, iFRET measurements, and cell viability testing with MTT method. The applicability of the switching DNA aptamer for the analysis of real samples, obtained by lysis of SW480 cells, was also tested. The proposed Apt(ATP) may be considered as a viable candidate for utilization in measurements of dynamic ATP level modulation in cells in different stages of cancer development and testing of new drugs in pharmacological studies.Graphical abstract

Regulatory Effect of Astragaloside A on Proliferation and Apoptosis of Human Colon Cancer SW480 Cell Line

Regulatory Effect of Astragaloside A on Proliferation and Apoptosis of Human Colon Cancer SW480 Cell Line

N-Acetylcysteine Serves as Substrate of 3-Mercaptopyruvate Sulfurtransferase and Stimulates Sulfide Metabolism in Colon Cancer Cells.

Hydrogen sulfide (H2S) is an endogenously produced signaling molecule. The enzymes 3-mercaptopyruvate sulfurtransferase (MST), partly localized in mitochondria, and the inner mitochondrial membrane-associated sulfide:quinone oxidoreductase (SQR), besides being respectively involved in the synthesis and catabolism of H2S, generate sulfane sulfur species such as persulfides and polysulfides, currently recognized as mediating some of the H2S biological effects. Reprogramming of H2S metabolism was reported to support cellular proliferation and energy metabolism in cancer cells. As oxidative stress is a cancer hallmark and N-acetylcysteine (NAC) was recently suggested to act as an antioxidant by increasing intracellular levels of sulfane sulfur species, here we evaluated the effect of prolonged exposure to NAC on the H2S metabolism of SW480 colon cancer cells. Cells exposed to NAC for 24 h displayed increased expression and activity of MST and SQR. Furthermore, NAC was shown to: (i) persist at detectable levels inside the cells exposed to the drug for up to 24 h and (ii) sustain H2S synthesis by human MST more effectively than cysteine, as shown working on the isolated recombinant enzyme. We conclude that prolonged exposure of colon cancer cells to NAC stimulates H2S metabolism and that NAC can serve as a substrate for human MST.

Evaluation of the Cytotoxic Effect of Hydroxypyridinone Derivatives on HCT116 and SW480 Colon Cancer Cell Lines

According to the literature, iron chelators have been used to inhibit tumor cell proliferation. Hydroxypyridinones, due to easy derivatization and high affinity for iron, have been suggested as an attractive target for the development of iron scavenging ligands. N-arylhydroxypyridinone derivatives as iron chelators have been previously designed and synthesized, and the present study is performed in order to evaluate the antitumor efficacy of these compounds,. Four derivatives of hydroxypyridinone were tested against HCT116 and SW480 colon cancer cell lines for 48 h using MTT assay. One compound (3-hydroxy-2-methyl-1-phenylpyridin-4(1H)-one, PMPO) showed the maximum cytotoxic activity on both HCT116 and SW480 cancer cells with IC50 = 243 and 180 μmol, respectively, for 48 h treatment. The obtained results demonstrated that various concentrations of test compounds exhibited significant reduction of the cell viability (P < 0.05) in a concentration dependent manner. Our findings indicate that the proposed hydroxypyridinone derivatives can be considered as a new option for the treatment of colon cancer.

Resveratrol inhibits the invasion and metastasis of colon cancer through reversal of epithelial‑ mesenchymal transition via the AKT/GSK‑3β/Snail signaling pathway.

The identification of safe and effective drugs that inhibit tumor invasion and metastasis is required to improve the clinical outcome of patients with colon cancer. The present study aimed to investigate the inhibitory effects and possible mechanisms of action of resveratrol against the invasion and metastasis of colon cancer. AKT1-knockdown SW480 and SW620 colon cancer cells were used to detect the effects of resveratrol on cell invasion and metastasis, as well as changes in the expression of epithelial-mesenchymal transition (EMT) markers and serine/threonine kinase (AKT)/glycogen synthase kinase (GSK)-3β/Snail signaling pathway-related molecules in vitro. Furthermore, nude mice were inoculated with SW480 cells in the tail vein to establish an in vivo lung metastasis model of colon cancer, to investigate the effects of resveratrol on lung metastasis in colon cancer. The results revealed that resveratrol treatment and AKT1 knockdown significantly inhibited cell migration and invasion in colon cancer, and markedly increased E-cadherin expression and decreased that of N-cadherin, phospho (p)-AKT1, p-GSK-3β, and Snail in colon cancer both in vitro and in vivo. Furthermore, the effects of resveratrol were significantly weaker in the AKT1-knockdown cells. In conclusion, resveratrol may suppress the invasion and metastasis of colon cancer through reversal of EMT via the AKT/GSK-3β/Snail signaling pathway. AKT1 may therefore be a key regulator of EMT in colon cancer cells and a potential therapeutic target for this disease.

Structural elucidation of an acidic polysaccharide from Citrus grandis ‘Tomentosa’ and its anti-proliferative effects on LOVO and SW620 cells

Citrus grandis ‘Tomentosa’ (CGT) which is a region-famous medicinal and edible plant contains plentiful bioactive polysaccharides, however, its chemical structures and specific bioactivities still need to be further explored. In the present study, an acidic polysaccharide (CGTP-AP) was extracted and purified from the pulps of CGT. The structure elucidation and anti-cancer activity of CGTP-AP were investigated. Structure characterization indicated that CGTP-AP was a homogeneous heteropolysaccharide composed of arabinose, galactose and galacturonic acid in a molar ratio of 2.45:1:2.77, with an average molecular weight of 2721.68 kDa. Partial acid hydrolysis, methylation and NMR spectrometry revealed that the backbone of CGTP-AP mainly composed of (1 → 4)-α-D-galacturonan, while the branch principally consisted of (1 → 5)-α-L-Araf. In addition, CGTP-AP exhibited effective anti-proliferation against colon cancer cells LOVO and SW620 cells in dose- and time-dependent manners, with IC50 values of 5.55, 4.35 and 3.52 mg mL−1 after 24, 48 and 72 h, and 5.33, 3.63 and 2.97 mg mL−1 after 24, 48 and 72 h, respectively. This study indicated that CGTP-AP might be utilized as a promising food supplement for the patients of colon disorders.

Proteomic screen with the proto-oncogene beta-catenin identifies interaction with Golgi coatomer complex I

Beta-catenin is well-known as a key effector of Wnt signalling and aberrant expression is associated with several human cancers. Stabilisation of and atypical subcellular localisation of beta-catenin, regulated in part through specific protein-protein interactions has been linked to cancer development, however the mechanisms behind these pathologies is yet to be fully elucidated. Affinity purification and mass spectrometry were used to identify potential β-catenin interacting proteins in SW480 colon cancer cells. Recombinant β-catenin constructs were used to co-isolate interacting proteins from stable isotope labelled cells followed by detection using mass spectrometry. Several known and new putative interactors were observed. In particular, we identified interaction with a set of coatomer complex I subunits implicated in retrograde transport at the Golgi, and confirmed endogenous interaction of β-catenin with coatomer subunit COPB using immunoprecipitation assays and immunofluorescence microscopy. These observations suggest a hitherto unrecognised role for β-catenin in the secretory pathway and warrant further functional studies to unravel its activity at this cellular location.

Abstract 4962: Repurposing bazedoxifene to suppress gastrointestinal cancer growth

Abstract The interleukin (IL)6 family of inflammatory cytokines, characterized by the shared use of the gp130 receptor, supports the progression of solid cancers. Specifically, gp130 signaling becomes rate limiting for the growth of gastrointestinal tumors arising from oncogenic driver mutations in Apc, but remains dispensable for the homeostatic renewal of the gastrointestinal mucosa. In the present study we investigate the effects of the selective estrogen receptor modulator bazedoxifene, clinically approved for the treatment of osteoporosis, as a putative small molecule inhibitor of gp130 receptor oligomerization and activation. We assessed the effects of bazedoxifene on IL6 and IL11-dependent STAT3 activation and cell proliferation in gastric and colon cancer cell lines. In patient-derived colon cancer organoid cultures, we assessed bazedoxifene effects on IL11-dependent growth. The gp130Y757F, Lgr5CreERT2- or Cdx2CreERT2-driven Apc flox mouse models of gastrointestinal cancer, were used to examine the effects of bazedoxifene on tumour growth. In silico modeling suggests that the inhibitory activity of bazedoxifene is due to its ability to mimic the interaction between the Site III interface of gp130 receptor and the tryptophan-157 in IL6, or tryptophan-168 in IL11 cytokine. We show that bazedoxifene interferes with IL6 and IL11-dependent proliferation of BAF/03 cells. Bazedoxifene suppresses gastric tumor growth in gp130Y757F mice irrespective of their gender, in which tumors arise through excessive IL11-dependent STAT3 signaling. Strikingly, in sporadic colon cancer models based on aberrant activation of the β-catenin/canonical WNT pathway, bazedoxifene treatment also reduces tumor burden in mice following conditional ablation of the Apc suppressor gene using the Lgr5CreERT2- or Cdx2CreERT2-driver alleles. Consistent with our observation that nuclear accumulation of β-catenin remains unaffected in colonic tumors of bazedoxifene-treated mice, bazedoxifenetreatment of human SW480 colon cancer cells harboring mutant APC did not affect canonical WNT signaling, but suppressed IL11-dependent STAT3 signaling. Our findings provide compelling proof-of-concept for the repurposing of bazedoxifene for the treatment of gastrointestinal cancers that arise from bona fide oncogenic driver mutations. Note: This abstract was not presented at the meeting. Citation Format: Pathum Thilakasiri, Tracy Nero, Michael W. Parker, Matthias Ernst, Ashwini L. Chand. Repurposing bazedoxifene to suppress gastrointestinal cancer growth [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 4962.

Abstract 5033: The role of the epigenetic regulator SATB2 in colon cancer progression

Abstract SATB2 is a nuclear matrix-associated transcription factor that orchestrates gene expression by regulating higher-order chromatin structure. Using antibody-based screening of 48 normal human tissues and 20 cancer types, we identified SATB2 as a protein almost exclusively expressed in gastrointestinal tissue. We confirmed this observation at the mRNA level in 10,533 human tissue samples. Differential expression of SATB2 was observed in colorectal cancer, with loss of expression occurring along the adenoma-carcinoma sequence. Additionally, SATB2 expression was markedly decreased in metastatic SW620 colon cancer cells and siRNA knockdown of SATB2 expression in parental SW480 cells increased their growth and migratory capacity. Ectopic expression of SATB2 in the metastatic variant reversed the observed phenotype. Using tissue microarray and automated image analysis of SATB2 expression in colorectal cancers (n=309), SATB2 was demonstrated by multivariate Cox regression analysis to be an independent predictor of disease-specific survival (HR=0.52, 95% CI 0.32-0.83, p=0.006). SATB2 mRNA levels were examined in a second cohort (n=290) and again, SATB2 was demonstrated to be an independent predictor of disease-specific survival (HR=0.40, 95% CI 0.18-0.92, p=0.031). Interestingly, in colorectal cancer patients, SATB2 levels significantly correlated with CD3+ T-cell infiltrates in the tumours (p=0.006) and inversely correlated with COX2 expression (p=0.019). In two independent colorectal cancer cohorts (n=467), SATB2-low tumours were found to be significantly enriched in the CMS1 (immune-related) subtype (p&amp;lt;0.001). Gene set enrichment analysis revealed that SATB2 low tumours demonstrated altered immune signalling with significant increases in IFNγ (p=0.001), IL6 (p=0.001), IL8 (p&amp;lt;0.001) TFGβ (p&amp;lt;0.001), which was mirrored by in vitro manipulation of SATB2 expression in colon cancer cells. Furthermore, in a longitudinal cohort of patients with ulcerative colitis, we observed a significant correlation between loss of SATB2 expression and occurrence of future cancers (p=0.013). We therefore postulate that SATB2 acts as a master regulator of the inflammatory response in the gut and loss of expression is significantly associated with the progression of colorectal cancer. Citation Format: Donal J. Brennan, Kirsha Naicker, Sudipto Das, Bruce Moran, Rut Klinger, Fredrik Ponten, Stephen Hewitt, Karin Jirström, Annette T. Byrne, Jacintha O'Sullivan, William M. Gallagher, Darran P. O'Connor. The role of the epigenetic regulator SATB2 in colon cancer progression [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 5033.

Effects and Mechanism of Tanshinone II A in Proliferation, Apoptosis, and Migration of Human Colon Cancer Cells.

BackgroundThe aim of this study was to explore the effect and mechanism of tanshinone II A on proliferation, apoptosis, and migration of human colon cancer cells.Material/MethodsCCK-8 approach was carried out to evaluate proliferation after applying various levels of tanshinone II A to SW620 colon carcinoma cells. Flow cytometry (FC) was used to assess apoptosis. Transwell assay was performed to assess invasion in vitro, and the wound-healing assay was applied to assess migration. Western blot analysis was performed to evaluate translation of mTOR, while RT-PCR was carried out to assess transcription of VEGF.ResultsCCK-8 assay showed that tanshinone II A inhibited SW620 proliferation in comparison to the control group subsequent to 24 h, 48 h, and 72 h (P<0.001). FC revealed that tanshinone II A promoted SW620 apoptosis (P<0.001). The cell migration test revealed that the migration index of cells receiving tanshinone II A decreased. mTOR translation as well as VEGE transcription in cells receiving tanshinone II A was noticeably prohibited compared to control group (P<0.001).ConclusionsTanshinone II A is able to inhibit proliferation and migration of human colon cancer SW620 cells and promoted cell death. Its mechanism may be by downregulation of mTOR protein and VEGF mRNA.

Combined blockade of MEK and PI3KCA as an effective antitumor strategy in HER2 gene amplified human colorectal cancer models

BackgroundTargeting the epidermal growth factor receptor (EGFR) either alone or in combination with chemotherapy is an effective treatment for patients with RAS wild-type metastatic colorectal cancer (mCRC). However, only a small percentage of mCRC patients receive clinical benefits from anti-EGFR therapies, due to the development of resistance mechanisms. In this regard, HER2 has emerged as an actionable target in the treatment of mCRC patients with resistance to anti-EGFR therapy.MethodsWe have used SW48 and LIM1215 human colon cancer cell lines, quadruple wild-type for KRAS, NRAS, BRAF and PI3KCA genes, and their HER2–amplified (LIM1215-HER2 and SW48-HER2) derived cells to perform in vitro and in vivo studies in order to identify novel therapeutic strategies in HER2 gene amplified human colorectal cancer.ResultsLIM1215-HER2 and SW48-HER2 cells showed over-expression and activation of the HER family receptors and concomitant intracellular downstream signaling including the pro-survival PI3KCA/AKT and the mitogenic RAS/RAF/MEK/MAPK pathways. HER2-amplified cells were treated with several agents including anti-EGFR antibodies (cetuximab, SYM004 and MM151); anti-HER2 (trastuzumab, pertuzumab and lapatinib) inhibitors; anti-HER3 (duligotuzumab) inhibitors; and MEK and PI3KCA inhibitors, such as refametinib and pictilisib, as single agents and in combination. Subsequently, different in vivo experiments have been performed. MEK plus PI3KCA inhibitors treatment determined the best antitumor activity. These results were validated in vivo in HER2-amplified patient derived tumor xenografts from three metastatic colorectal cancer patients.ConclusionsThese results suggest that combined therapy with MEK and PI3KCA inhibitors could represent a novel and effective treatment option for HER2-amplified colorectal cancer.

Phytochemical Evaluation, Antimicrobial and Anticancer Properties of New “Oligo Grapes” Supplement

This research presents complete data about phytochemical analysis and biological activities of a new dietary supplement for commercial use “Oligo Grapes” (OG), and its individual constituents: lyophilized pomace (LP), clay supplemented with red wine extract, lyophilized pomace ethanolic extract (PE), and red wine extract. OG, as a commercially available food supplement, has not been previously tested for its biological activity. Now we want to present new data about its phytochemical screening, antioxidant and antibacterial activities, and anticancer properties with respect to its cytotoxicity and effects on redox status in colon cancer cell lines. The tested extracts expressed strong antibacterial activity against Proteus mirabilis and Proteus mirabilis ATCC 12453, where the synergy of contents inside the supplement demonstrated a higher influence on the bacteria than its separately tested constituents. Among the investigated extracts, PE, as the extract with the highest phenolics concentration, had remarkable cytotoxic activity on HCT-116 and SW-480 colon cancer cells. Also, the treatments modulated redox status in the investigated cancer cells, by inducing oxidative and nitrosative stress, which could be one of the preferred mechanisms of the anticancer action. Based on the achieved antimicrobial and anticancer properties, there is a need for producing different food supplements and nutritional products originating from grapes. In this respect, the food supplement OG and its health benefits deserve scientific attention and further research.

166-LB: Canagliflozin but Not Empagliflozin Activates AMPK and Suppresses Colon Cancer Cell Growth

Background: Recent increase in the relative proportion of cancer death in diabetic patients has become a major issue. A hypoglycemic agent, sodium-glucose cotransporter (SGLT) 2 inhibitor has attracted attention because of its potency in reducing cardiovascular risks. Several lines of evidence has emerged suggesting a potential anti-oncotic effect. Here we address potential effects of canagliflozin (Cana) on human colon cancer cell growth and underlying mechanisms and compared with empagliflozin (Empa). Material and Methods: We examined the effects of Cana and Empa on 1) cell proliferation using colon cancer cell lines, SW480 and HCT116 by MTT assay, 2) DNA synthesis by flow cytometry-based analysis of bromodeoxyuridine (BrdU) uptake. We further examined 3) the expression of SGLT1 and 2 in these cells and 4) investigated molecules of signal transduction pathways with established roles in cancer cell growth such as AMPK, 4EBP1, mTOR by western blotting. Finally, we analyzed 5) time-dependent changes of metabolomic signatures after the treatment with Cana. Results: While SGLT1 and 2 mRNA and protein expression was detected in both cell lines, Cana but not Empa significantly inhibited cell proliferation and in a dose-dependent manner. Cana suppressed BrdU positive cells and enhanced AMPK phosphorylation with a consequent suppression of 4EBP1 and mTOR activity. Metabolomic profiling revealed that, as a function of time, intermediate metabolites of glycolysis especially phosphoenolpyruvate was decreased with an acute rise in ADP and gradual decrease in ATP and GSSG. Conclusion: We demonstrate that Cana deprives human colon cancer cells of energy and anti-oxidative capacity possibly through an inhibition of SGLT1-mediated glucose influx, and thereby activates AMPK and inhibits cancer cell growth. Our data herein suggest a potential beneficial effect of Cana in suppressing colon cancer progression especially in patients with diabetes. Disclosure T. Yagi: None. T. Tanaka: Research Support; Self; Japan Society for the Promotion of Science. Speaker's Bureau; Self; AstraZeneca, Boehringer Ingelheim Pharmaceuticals, Inc., Daiichi Sankyo Company, Limited, Kissei Pharmaceutical Co., Ltd., Kowa Pharmaceutical Europe Co. Ltd., Mitsubishi Tanabe Pharma Corporation, Novartis Pharmaceuticals Corporation, Taisho Toyama Pharmaceutical Co. Ltd., Takeda Pharmaceutical Company Limited. E. Kubota: None. H. Koyama: None. T. Guo: None. K. Ogawa: None. D. Aotani: None. H. Kataoka: None.

Pharmacological effects of the simultaneous and sequential combinations of trifluridine/tipiracil (TAS-102) and 5-fluorouracil in fluoropyrimidine-sensitive colon cancer cells.

The aim of this study was to investigate possible synergistic effects in vitro of trifluridine/tipiracil (TAS-102) and 5-fluoruracil (5-FU) on fluoropyrimidine-sensitive colon cancer cell lines of different mutational status in order to build a rational basis for the future use of this combination therapy in adjuvant settings or as a first-line treatment for metastatic disease. Proliferation assays were performed on HT-29 (B-raf mutated), SW-620 (ras mutated), and Caco-2 (wild type) colon cancer cell lines exposed to 120-h treatments of 5-FU, TAS-102 and their different combination schedules (simultaneous, sequential and reverse) at equimolar and non-equimolar ratios. The synergistic, additive and antagonistic effects of 5-FU and TAS-102 were determined by the combination index (CI) and dose reduction index (DRI). Our preclinical in vitro results may suggest an apparently counterintuitive but strongly synergistic combination of 5-FU and TAS-102 in fluoropyrimidine-sensitive colon cancer cells allowing a marked theoretical reduction in the administered doses of both drugs. In particular, this association seems to be highly effective in wild-type colon cancer cells, both in sequential and simultaneous schedules. Together, these data may build a rational basis for the future use of TAS-102 combined with 5-FU in adjuvant settings, or as a first-line treatment for metastatic disease.

Inhibitory effects of petasin on human colon carcinoma cells mediated by inactivation of Akt/mTOR pathway.

Background:Colorectal cancer is the third most common cancer worldwide and still lack of effective therapy so far. Petasin, a natural product found in plants of the genus Petasites, has been reported to possess anticancer activity. The present study aimed to investigate the anticolon cancer activity of petasin both in vitro and in vivo. The molecular mechanism of petasin was also further explored.Methods:Caco-2, LoVo, SW-620, and HT-29 cell lines were used to detect the inhibitory effect of petasin on colon cancer proliferation. Cell viability was determined using the MTT (3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide) assay. Cell apoptosis was analyzed by flow cytometry. Hoechst 33258 staining was used to visualize morphological changes. Cell migration was assessed using a wound-healing migration assay, and cell invasion was investigated using Transwell chambers. Western blotting assays were employed to evaluate the expression levels of proteins in the protein kinase B/mammalian target of rapamycin (Akt/mTOR) signaling pathway. Finally, in vivo activity of petasin was evaluated using the SW-620 subcutaneous tumor model established in Balb/c nude mice. Twelve rats were randomly divided into control group and 10 mg/kg petasin group. The tumor volume was calculated every 7 days for 28 days. Terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) assay was performed to assess the apoptotic effect of petasin. Differences between two groups were assessed by analysis of independent-sample t tests.Results:Petasin significantly inhibited the proliferation of human colon carcinoma cell lines, induced apoptosis, and suppressed migration and invasion in SW-620 cells. Western blotting results showed that petasin decreased the phosphorylation of Akt (1.01 ± 0.16 vs. 0.74 ± 0.06, P = 0.042), mTOR (0.71 ± 0.12 vs. 0.32 ± 0.11, P = 0.013), and P70S6K (1.23 ± 0.21 vs. 0.85 ± 0.14, P = 0.008), elevated the expression of caspase-3 (0.41 ± 0.09 vs. 0.74 ± 0.12, P = 0.018) and caspase-9 (1.10 ± 0.27 vs. 1.98 ± 0.22, P = 0.009), decreased the Bcl-2 protein (2.75 ± 0.47 vs. 1.51 ± 0.36, P = 0.008), downregulated the expression of matrix metalloproteinase (MMP)-3 (1.51 ± 0.31 vs. 0.82 ± 0.11, P = 0.021) and MMP-9 (1.56 ± 0.32 vs. 0.94 ± 0.15, P = 0.039) in SW-620 cell. In vivo, 10 mg/kg petasin inhibited tumor growth in Balb/c nude mice (924.18 ± 101.23 vs. 577.67 ± 75.12 mm3 at day 28, P = 0.001) and induced apoptosis (3.6 ± 0.7% vs. 36.0 ± 4.9%, P = 0.001) in tumor tissues.Conclusions:Petasin inhibits the proliferation of colon cancer SW-620 cells via inactivating the Akt/mTOR pathway. Our findings suggest petasin as a potential candidate for colon cancer therapy.

Ethyl Acetate Extracts of Two Artemisia Species: Analyses of Phenolic Profile and Anticancer Activities Against SW-480 Colon Cancer Cells

Because Artemisia vulgaris L. and Artemisia alba Turra are traditional medicinal plants used for the treatment of different diseases, we evaluated the cytotoxic/apoptotic activity of ethyl acetate extracts from these natural products against human colon cancer cells SW-480. The extracts contained a large amount of the total polyphenols and flavonoids. The phenolic profile showed the presence of phenolic acids (gallic, p-coumaric, vanillic, and ferulic acids) and flavonoids (rutin, myricetin, luteolin, quercetin, and apigenin). 3-[4,5-Dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide assay indicated that antiproliferative activities of both A. vulgaris and A. alba extracts increased with the extension of time exposure, with decreasing IC50 values. Mitomycin C (MMC) alone had antiproliferative activity, but in combination with plant extracts caused stronger effect with lower IC50 values. Flow cytometry analyses showed that A. alba extract induced higher percentage of SW-480 cells in the early stage of apoptosis (33.5 ± 1.6 vs 0.7 ± 0.1, P &lt; 0.05), whereas the A. vulgaris extract significantly increased the percentage of cells in necrosis (82.4 ± 5.0 vs 53.9 ± 2.3, P &lt; 0.05). In conclusion, A. alba extract can be considered a potential source of bioactive components with anticancer activity or be used as a dietary food supplement or supplement to chemotherapy due to its synergistic effect with the MMC.

Synthesis of luteolin loaded zein nanoparticles for targeted cancer therapy improving bioavailability and efficacy

A naturally occurring flavonoid, Luteolin possess wide range of pharmaceutical activities from anti-inflammatory to anticancer effect. Luteolin inhibits cell proliferation by arresting cell cycle pathways and triggers apoptotic cell death. Poor aqueous solubility of luteolin limits its oral bioavailability. Therefore, to improve the oral bioavailability of luteolin by use of biodegradable protein zein and sodium caseinate as nanocarrier. The nanoparticles were prepared by simple precipitation process. The resulting nanoparticles had mean size of 200–300 nm with negative zeta potential with 92% encapsulation efficiency and luteolin release at intestinal pH. Anti-oxidant activity of loaded molecule with respect to parent molecule significantly indicate the free radical scavenging potential of luteolin increases after encapsulation in zein nanoparticles. Luteolin loaded zein nanoparticles showed enhanced cytotoxicity against SW480 colon cancer cells and induces apoptosis. In this paper, we have shown the potential of luteolin loaded zein nanoparticles for treating colon cancer which could serve as promising strategy to develop oral formulation of this water insoluble drug enhancing its bioavailability.