Human Colon Adenocarcinoma Cancer Research Articles

Design, synthesis and biological evaluation of 1,2,4-oxadiazole linked 1,2,4-thiadiazole-pyrimidines as anticancer agents

Abstract A novel library of 1,2,4-oxadiazole linked 1,2,4-thiadiazole-pyrimidines (10a-j) were designed and synthesized. All compounds were characterized by FTIR, NMR and high resolution Mass analysis. Further, these compounds were screened for their anticancer activity against a panel of human cancer cell lines such as MCF-7 (human breast cancer), A-549 (human non -small cell lung cancer), Colo-205 (human adenocarcinoma colon cancer) and A2780 (human ovarian cancer) by employing MTT assay. The clinical drug (etoposide) used as positive control. Among all synthesized compounds, five compounds 10b, 10c, 10d, 10i and 10j were possessed more promising anticancer activity on all cell lines.

Combinatorial Cytotoxic Effects of Gelam Honey and 5-Fluorouracil against Human Adenocarcinoma Colon Cancer HT-29 Cells In Vitro

Combination of natural products with chemodrugs is becoming a trend in discovering new therapeutics approach for enhancing the cancer treatment process. In the present study, we aimed to investigate the cytotoxic and apoptosis induction of Gelam honey (GH) combined with or without 5-Fluorouracil (5-FU) on HT-29 cells. The cell viability was determined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay to assess cytotoxicity. Morphological changes and apoptosis were determined by the inverted microscope, Annexin V-FITC, and DNA fragmentation via flow cytometric analysis, respectively. Our results demonstrate that combined treatment revealed a remarkable and concentration-dependent cytotoxic effect on HT-29 cells in comparison with GH and 5-FU alone. Flow cytometry analysis showed that early apoptosis event was more pronounced in combined treatment. In addition, compared to 5-FU alone, apoptosis of HT-29 cells treated with combinations of GH and 5-FU demonstrated increasing percentages of fragmented DNA. Our results suggest that GH has a synergistic cytotoxic effect with 5-FU in HT-29 cell lines in vitro. Although the actions of the molecular mechanisms are not yet clear, the results reveal that the combination of GH and 5-FU could have the potential as a therapeutic agent.

PO-049 Quantitative proteomics unveils key mitotic regulators involved in the transition to androgen independent prostate cancer

IntroductionMarine invertebrate organisms have received great attention from the scientific community due to its richness in new bioactive compounds with potential anti-proliferative and anticancer activities. In the present study, the cytotoxic efficacy of four marine invertebrate-extracts against human adenocarcinoma colon cancer cells was investigated.Material and methodsHT29, SW480 and HGUE-C-1 human colon adenocarcinoma cell lines were cultured and exposed to 4 marine invertebrate extracts at different concentrations for 24 hour. The anti-proliferative properties were examined by MTT assay. Analysis of apoptosis and cell cycle stages were performed with the Muse Cell Analyzer (Millipore, Hayward, CA, USA). Mitochondrial functionality was determined by the ratio of MitoTracker Red CMXRos and MitoTracker Green FM fluorescent probes (Molecular Probes). Measurement of lactate dehydrogenase (LDH) was performance by a colorimetric assay (Roche Diagnostic, Germany).Results and discussionsThe four marine invertebrate extracts exhibited pronounced cytotoxic effects in the three cancer cell lines used. In this study, CR, PS, NA and NB extracts were able to modify the cell cycle of HGUE-C-1, HT-29 and SW-480 cells, by increasing cell population in the subG1 and G2/M phase in a dose-dependent manner. Results confirmed that NA, NB and CR extracts induced an increase in the percentage of cell population in early apoptosis by Annexin V staining. PS extract on the contrary, raised the late apoptotic population at different concentrations. Mitochondrial membrane depolarization rate, as indicator of apoptosis, was induced significantly by CR, NA and NB extracts in the three cell models used. In contrast PS reduced this parameter in a dose-dependent manner.Finally, the release of the enzyme lactate dehydrogenase (LDH) confirmed a cell death by a necrotic pathway for PS extract.ConclusionOur results strongly suggest that NB, NA and CR extracts exhibit antiproliferative and pro-apoptotic properties in colon cancer cell lines concomitantly to a decreased mitochondrial membrane potential. In contrast, PS extract shows an antiproliferative effect mediated by necrosis in the same cell models. This preliminary study suggests that these extracts present pharmacological potential. Further investigations to determine the responsible compounds and the underlying mechanism are currently ongoing.

Photo-catalytic, anti-bacterial, and anti-cancer properties of phyto-mediated synthesis of silver nanoparticles from Artemisia tournefortiana Rchb extract

Metal nanoparticles have largely been investigated due to their potential medicinal activities. This study demonstrates the biological properties of green-synthesized silver nanoparticles (AgNPs) by using Artemisia tournefortiana Rchb ethanol extract. Instrumentations such as ultraviolet-visible spectra analysis, high-resolution transmission electron microscopy, scanning electron microscopy, energy-dispersive X-ray analysis, X-ray diffraction, and Fourier transform infrared spectroscopy were used to reveal the synthesized AgNPs. Microscopic results showed that the particles were mostly spherical in shape, having an average diameter of 22.89±14.82nm. The antibacterial activity of the phyto-fabricated AgNPs was investigated by the determination of minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC). The in vitro cytotoxicity effect was investigated against normal human embryonic kidney (HEK293) cells and human colon adenocarcinoma cancer (HT29) cells. The apoptotic cells were identified by annexin V/PI FITC staining, and morphological assessment. The expressions of Bax and Bcl2 were evaluated by quantitative real time PCR method. The phyto-synthesized AgNPs have shown increased cell apoptosis and demonstrated dose-dependent cytotoxicity in HT29 cancer cells. Moreover, the photocatalytic activity of the phyto-synthesized AgNPs was evaluated by degradation of Coomassie Brilliant Blue G-250 under UV light exposure and these fabricated Ag nanoparticles demonstrated efficacy in degrading the dye within 60min. Overall, the present results highlighted the antibacterial and anticancer properties of fabricated AgNPs, suggesting that phyto-synthesized silver nanoparticles could possess potent anti-pathogenic bacteria and anti-colon cancer activities.

Abstract 5437: A novel method to quantify gamma H2AX foci in circulating tumor cells in patients receiving chemotherapy for colorectal cancer

Abstract Phosphorylated H2AX (γH2AX) is a marker of DNA double-strand break damage and has been proposed as a pharmacodynamic biomarker following treatment with chemotherapy. γH2AX levels in circulating tumor cells (CTCs) may provide a sensitive direct marker for assessing drug effects in tumor tissue without the need for biopsy. In the present study, we established a novel assay to quantify changes in γH2AX in CTCs from patients with metastatic colorectal cancers undergoing treatment with Fulfox or Folfiri. γH2AX foci were initially assessed in human colon adenocarcinoma cancer cell lines (HT-29) treated with different concentrations of Oxaliplatin and SN-38 at different time points. Image capture was conducted using Confocal Microscope Leica SPE2 and γH2AX foci were analyzed with CellProfiler software. The highest number of γH2AX foci/nucleus was induced at 5uM for Oxaliplatin and 0.01uM for SN-38 and the peak of γH2AX foci/nucleus was observed between two-six hours post dose. To reproduce the analytical process for CTCs, we evaluated the γH2AX signal using both the CellSearch System (Janssen Diagnostics) and the DEPArray™ System (Silicon Biosystems). HT-29 cells were used, either as untreated controls or following 2 hours treatment with Oxaliplatin 5uM or SN-38 0.01uM and spiked in to healthy donor blood. Cells were defined as positive for γH2AX on the CellSearch according to a previously validated assay. For Oxaliplatin and SN-38 treated cells, 15.28% and 18.37% respectively were classified as positive for γH2AX, compared with 5.10% for untreated controls. However, using CellSearch, the fluorescent signal could not be quantified and we therefore repeated the experiment using the DEPArray platform. HT-29 cells were treated with SN-38 as above, and compared with an untreated control group using two different exposure times for fluorescein isothiocyanate-conjugated antibody (FITC): FITCI (100 ms and gain 1X) and FITCII (800 ms and gain 4X). The treated group showed a significantly increased intensity of FITC staining compared with the untreated control group: mean 363 vs mean 220 (p<0.0001) for FITCI, and mean 5521 vs mean 4365 (p<0.0040) for FITCII. The DEPArray system therefore has an advantage in being able to quantify differences in signal intensity caused by induction of γH2AX in CTCs and then select individual cells for further downstream analysis. This analysis is now being applied to patient samples collected pre and post-chemotherapy to evaluate the utility of the assay in a clinical setting. Citation Format: Matilde Saggese, Leah Ensell, Clare Vesely, Victoria Spanswick, Elena Peruzzi, Diana Cunati, Giulio Signorini, John Hartley, Hendrik Tobias Arkenau, Tim Meyer. A novel method to quantify gamma H2AX foci in circulating tumor cells in patients receiving chemotherapy for colorectal cancer. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 5437. doi:10.1158/1538-7445.AM2015-5437

Polydatin, a natural precursor of resveratrol, induces cell cycle arrest and differentiation of human colorectal Caco-2 cell

BackgroundHuman colon adenocarcinoma cells are resistant to chemotherapeutic agents, such as anthracyclines, that induce death by increasing the reactive oxygen species. A number of studies have been focused on chemo-preventive use of resveratrol as antioxidant against cardiovascular diseases, aging and cancer. While resveratrol cytotoxic action was due to its pro-oxidant properties. In this study, we investigate whether the Resveratrol (trans-3,5,49-trihydroxystilbene) and its natural precursor Polydatin (resveratrol-3-O-b-mono- D-glucoside, the glycoside form of resveratrol) combination, might have a cooperative antitumor effect on either growing or differentiated human adenocarcinoma colon cancer cells.MethodsThe polydatin and resveratrol pharmacological interaction was evaluated in vitro on growing and differentiated Caco-2 cell lines by median drug effect analysis calculating a combination index with CalcuSyn software. We have selected a synergistic combination and we have evaluated its effect on the biological and molecular mechanisms of cell death.ResultsSimultaneous exposure to polydatin and resveratrol produced synergistic antiproliferative effects compared with single compound treatment. We demonstrated that polydatin alone or in combination with resveratrol at 3:1 molar ratio synergistically modulated oxidative stress, cell cycle, differentiation and apoptosis. Worthy of note treatment with polydatin induced a nuclear localization and decreased expression of heat shock protein 27, and vimentin redistributed within the cell.ConclusionsFrom morphological, and biochemical outcome we obtained evidences that polydatin induced a transition from a proliferative morphology to cell-specific differentiated structures and caused human CaCo-2 cell death by induction of apoptosis. Our data suggest the potential use of polydatin in combination chemotherapy for human colon cancer.

Positional Isomers of Aspirin Are Equally Potent in Inhibiting Colon Cancer Cell Growth: Differences in Mode of Cyclooxygenase Inhibition

We compared the differential effects of positional isomers of acetylsalicylic acid (o-ASA, m-ASA, and p-ASA) on cyclooxygenase (COX) inhibition, gastric prostaglandin E2 (PGE2), malondialdehyde, tumor necrosis factor-alpha (TNF-α) levels, superoxide dismutase (SOD) activity, human adenocarcinoma colon cancer cell growth inhibition, cell proliferation, apoptosis, and cell-cycle progression. We also evaluated the gastric toxicity exerted by ASA isomers. All ASA isomers inhibit COX enzymes, but only the o-ASA exerted an irreversible inhibitory profile. We did not observe a significant difference between ASA isomers in their ability to decrease the in vivo synthesis of PGE2 and SOD activity. Furthermore, all isomers increased the levels of gastric and TNF-α when administered orally at equimolar doses. We observed a dose-dependent cell growth inhibitory effect; the order of potency was p-ASA > m-ASA ≈ o-ASA. There was a dose-dependent decrease in cell proliferation and an increase in apoptosis, with a concomitant Go/G1 arrest. The ulcerogenic profile of the three ASA isomers showed a significant difference between o-ASA (aspirin) and its two positional isomers when administered orally at equimolar doses (1 mmol/kg); the ulcer index (UI) for o-ASA indicated extensive mucosal injury (UI = 38), whereas m-ASA and p-ASA produced a significantly decreased toxic response (UI = 12 and 8, respectively) under the same experimental conditions. These results suggest that the three positional isomers of ASA exert practically the same biologic profile in vitro and in vivo but showed different safety profiles. The mechanism of gastric ulcer formation exerted by aspirin and its two isomers warrants a more detailed and thorough investigation.

Human Gut Flora-Fermented Nondigestible Fraction from Cooked Bean (Phaseolus vulgaris L.) Modifies Protein Expression Associated with Apoptosis, Cell Cycle Arrest, and Proliferation in Human Adenocarcinoma Colon Cancer Cells

Metabolism of the nondigested fraction (NDF) from common bean ( Phaseolus vulgaris L.) by the human gut flora (hgf) produces short-chain fatty acids (SCFAs) that may benefit cancer by reducing colorectal tumor risks. This paper reports the effect of fermentation products (FP) by hgf (FP-hgf) from NDF of cooked beans on survival and protein expression associated with apoptosis, cell cycle arrest, and proliferation in human adenocarcinoma colon cancer cells. FP-hgf was the only inoculum eliciting butyrate production after 24 h of NDF fermentation using different bacterial sources. FP-hgf inhibited HT-29 cell growth and modulated protein expression associated with apoptosis, cell cycle arrest, and proliferation, as well as morphological changes linked to apoptosis evaluated by TUNEL and hematoxylin and eosin stains, confirming previous results on gene expression. The current results suggest that fermentation of NDF from common beans can elicit beneficial chemoprotective effects in colon cancer by modulating protein expression in HT-29 cells.

Establishment of doxorubicin-resistant subline derived from HCT15 human colorectal cancer cells

Doxorubicin, one of the clinically most useful anticancer agents, is used alone or in combination with other drugs against a wide variety of tumors, recently. But cancer cells developed resistance to this agent in many ways. This resistance is an important limiting factor of doxorubicin for anticancer drug. We newly established doxorubicin-resistant HCT15/CL02 subline from parental HCT15 human adenocarcinoma colon cancer cells. HCT15/CL02 revealed resistance to doxorubicin about 85-fold of its parental cells, and it also revealed cross-vealed to actinomycin D, etoposide and vinblastine but not to cisplatin and tamoxifen. And verapamil, a reversal agent of multidrug-resistance (MDR) by P-glycoprotein, elevated the cytotoxicity of doxorubicin against both HCT15 and HCT15/CL02 cells. But the relative resistant rate was not reduced. Verapamil had no effects on the toxicity of cisplatin to the both cell lines. These results indicate that HCT15/CL02 cells have some functionally complex mechanisms for MDR.