DLD-1 Cell Lines Research Articles

A Preliminary Study on Anti-Colorectal Cancer Effect and Molecular Mechanism of Aegiceras Corniculatum Extract

Objective: To study the inhibitory effects on colorectal cancer (CRC) and the underlying mechanism of the petroleum ether extract of Aegiceras corniculatum leaves (PACL). Materials and Methods: The effect of PACL on the proliferation of CRC cell lines DLD-1, HT-29, and SW480 was measured by 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium assay and colony-forming assay. And then, a wound-healing assay was used to measure the migration ability of three CRC cells. The cell cycle and apoptosis of three CRC cells were measured by PI/RNase staining and annexin V-FITC/double staining, respectively, and the intrinsic apoptosis pathway was studied by the Western blot. The anti-CRC effect of PACL in vivo was evaluated by HT-29 xenograft zebrafish embryos. Results: PACL inhibited cell viability and proliferation in DLD-1, HT-29, and SW480 cells in a dose- and time-dependent manner. PACL can inhibit cell migration in DLD-1 and SW480 cells but not in the less mobile phenotype cell HT-29. PACL treatment resulted in cell cycle arrest of DLD-1 and HT-29 cells in the G2/M phase. Moreover, PACL can induce apoptosis in all three CRC cells, which may be achieved by regulating the intrinsic apoptosis pathway mediated by mitochondria and the endoplasmic reticulum. Interestingly, the tumor sizes were decreased after treatment with PACL and PACL combined with fluorouracil in HT-29 xenograft zebrafish embryos. Conclusions: These findings suggested that PACL may exert its anti-CRC effect by inducing apoptosis through the intrinsic apoptosis pathway and show a significant anti-CRC effect in vitro and in vivo, so it might be potentially developed as an anti-CRC agent.

DETERMINATION OF CYTOTOXIC EFFECT OF AMYGDALIN IN DLD-1 CELL LINE AND ANTICYTOTOXIC EFFECT IN CCD-18CO CELL LINE

Objective: Amygdalin, which is part of the aromatic cyanogenic glycoside group, is found in plant seeds such as apricot, peach, plum, apple, pear, and cherry. It has been shown that amygdalin has anti-tumor properties against many cancers such as colon, breast, and lung cancer. This study aimed to determine the cytotoxic and anticytotoxic effects of amygdalin in human colon cancer cells (DLD-1) and normal colonic epithelium (CCD-18Co) using the MTT (3-(4,5-dimethylthiazol-2-YL)-2,5-diphenyltetrazolium bromide) test. 
 Materials and Methods: DLD-1 and CCD-18Co cells were grown in flasks containing Roswell Park Memorial Institute-1640 and Eagle's Minimum Essential Medium, respectively. Both cell groups were treated with amygdalin concentrations of 100, 50, 25, 12.5, 6.25, 3.125, and 1.56 mM for 24 hours. Then, 20% MTT dye was added to the wells of the aspirated plates and incubated for 3 hours. After the reaction was stopped with pure DMSO at the end of the period, the absorbance values of the plates were read spectrophotometrically at a wavelength of 570 nm.
 Results: The percent viability values for the DLD-1 cell line were found to be between 48.3-71.6% and the IC50 value was calculated as 74.03 mM. The viability values for the CCD-18Co cell line after the amygdalin treatment ranged from 101.6 to 117.9%.
 Conclusion: While amygdalin showed a cytotoxic effect in the DLD-1 cell line, it showed an anticytotoxic effect in the CCD-18Co cell line. In our study, it was determined that amygdalin decreased the viability of DLD-1 cancer cells in a dose-dependent manner and did not show cytotoxic effects on CCD18-Co normal epithelial cells. More comprehensive controlled clinical trials are needed to demonstrate the feasibility of using amygdalin in combination with other anti-tumor drugs and to develop the artificial synthesis of the active ingredients in amygdalin in order to increase the anti-tumor activities of these drugs.

Use of a Foamy-Virus Vector System to Produce an 'Off-the-Shelf' Fcγ-CR-T Cell Product for the Treatment of Hematological and Solid Tumour Malignancies

Introduction Chimeric antigen receptor (CAR) T cells have transformed the field of cancer immunotherapy. Other approaches, such as the use of Fc gamma chimeric receptor (Fcγ-CR)-T cells have further expanded the applicability of such therapies in both solid and liquid tumours with the added benefit of tackling some of the hurdles associated with CAR-T therapies (such as loss or down-regulation of the target antigen). Previous studies have generated Fcγ-CR-T cells from autologous cells, which carry several limitations (prolonged production time, costly, manufacturing failure), whereas the use of Lentiviral vectors (LV) is endowed with extra limitations (packaging limits and mutagenesis risk). Our group has developed an in-house Fcγ-CR-T cell product, using a safer to LV, foamy virus (FV) vector and T cells from healthy donors. Materials We constructed FV vectors expressing the CD16 V158, which has higher Fc binding and is associated with higher tumour killing, the T cell stimulatory molecule CD3ζ and the costimulatory molecule CD28 and an EFP1a promoter. 2nd generation LV vector backbones were purchased from a commercial vendor. Peripheral blood from healthy individuals were used as T cell sources. T cells were activated by anti-CD3/CD28 beads and transduced with CD16-CR, LV or FV vectors. Transduction efficiency was assayed by flow cytometry (FCM) using an anti-CD16 antibody (Ab), on day 3. The human cell lines Raji, Panc01 and DLD-1 were used for functional assays, in the presence of the Abs Rituximab (anti-CD20) and Cetuximab (anti-EGFR), respectively, at a concentration of 0.1 ug/ml. The CR's Ab-binding capacity was assessed by incubating the CD16-CRs with the Abs for 30mins, followed by an anti-human Ig PE antibody and the related fluorescence intensity was assessed. To determine whether Ab binding to CD16-CR can promote aggregation of effector and target cells, CFSE-labelled target cells were mixed with Ab-coated CD16-CRs for 60 mins and the formation of CFSE-PE doublets was assessed by FCM. Their cytotoxic effect was evaluated against the CFSE-labelled Raji or DLD-1 or Panc01 cells at different ratios (5:1, 10:1) for 18 hours, in the presence of Rituximab (0.1ug/ml) or Cetuximab (0.1ug/ml), respectively. The % of live cells was assessed by flow cytometry and calculated as: [1-live targets (sample)/live targets (control)]x100. Results LV and FV vector titers were between 3-5x10^6 TU/ml and 4-5x10^6 TU/ml, respectively. Transduction efficiency ranged from 58.3-69.2% with FV vectors (MOI 3-5) and 85.2-85.9% with LV vectors (MOI 10-20). The Ab-binding capacity of FV-CD16-CRs was determined to be 68.7% (SEM 2.3) and 71.3% (SEM 3.1), (n=3) for Rituximab and Cetuximab, respectively, whereas that of LV-CD16-CRs was 72.1 (SEM 3.3) and 76.5 (SEM 2.8), (n=3), respectively. Cell aggregation of effector and target cells, assessed as doublets, was: (i) 32%, 39% and 36% for FV-CD16-CRs and (ii) 26%, 31% and 29% for LV-CD16-CRs, coated with Rituximab and Cetuximab, respectively, and it was specific for the Raji, DLD-1 and Panc01 cells, respectively. Next, we assessed whether CD16-CR T cells were able to kill target cells in the presence of specific antibodies. Results showed that the FV-CD16-CR-induced % cell lysis was: (i) 26.3% and 51.5% at 5:1 and 10:1 ratio, respectively, in the presence of Rituximab and Raji cells, (ii) 41.5% and 57.7% at 5:1 and 10:1 ratio, respectively, in the presence of Cetuximab and DLD1 cells and (iii) 39.4% and 57.3% at 5:1 and 10:1 ratio, respectively, in the presence of Cetuximab and Panc01 cells. For LV-CD16-CRs the respective % lysis was comparable. More importantly, this lysis was shown to be specific (no lysis noted with untransduced T cells) and significantly lower in the absence of the antibodies. Conclusion Our group has developed for the first time a FV vector for the generation of CD16-CR T cells, with an efficient gene transfer to human T cells and with potent in vitro cytotoxic properties, similar to their LV-derived counterpart. Overall, we provide a proof of concept that allogeneic, in-house Fcγ-CR-T cells derived from a non-pathogenic viral backbone such as the FV, could be a safe, efficient and affordable alternative to LV-derived vectors for immunotherapy. Our future aim is to further modify these Fcγ-CR-T cells, by interfering with immune checkpoint molecules, in order to achieve better tumour penetration and tackle the anergy induced by the tumour microenvironment.

2.4 GHz Electromagnetic Field Influences the Response of the Circadian Oscillator in the Colorectal Cancer Cell Line DLD1 to miR-34a-Mediated Regulation.

Radiofrequency electromagnetic fields (RF-EMF) exert pleiotropic effects on biological processes including circadian rhythms. miR-34a is a small non-coding RNA whose expression is modulated by RF-EMF and has the capacity to regulate clock gene expression. However, interference between RF-EMF and miR-34a-mediated regulation of the circadian oscillator has not yet been elucidated. Therefore, the present study was designed to reveal if 24 h exposure to 2.4 GHz RF-EMF influences miR-34a-induced changes in clock gene expression, migration and proliferation in colorectal cancer cell line DLD1. The effect of up- or downregulation of miR-34a on DLD1 cells was evaluated using real-time PCR, the scratch assay test and the MTS test. Administration of miR-34a decreased the expression of per2, bmal1, sirtuin1 and survivin and inhibited proliferation and migration of DLD1 cells. When miR-34a-transfected DLD1 cells were exposed to 2.4 GHz RF-EMF, an increase in cry1 mRNA expression was observed. The inhibitory effect of miR-34a on per2 and survivin was weakened and abolished, respectively. The effect of miR-34a on proliferation and migration was eliminated by RF-EMF exposure. In conclusion, RF-EMF strongly influenced regulation mediated by the tumour suppressor miR-34a on the peripheral circadian oscillator in DLD1 cells.

Effects of Sulfur Containing Glycine Imine Derivatives Compounds on Multidrug Resistance Proteins (MRPs) and Apoptosis Mechanism in MCF-7 and DLD-1 Cell Lines

Effects of Sulfur Containing Glycine Imine Derivatives Compounds on Multidrug Resistance Proteins (MRPs) and Apoptosis Mechanism in MCF-7 and DLD-1 Cell Lines

Establishment of In Vitro and In Vivo Anticolorectal Cancer Efficacy of Lithocholic Acid-Based Imidazolium Salts.

Imidazolium salts (IMSs) are the subject of many studies showing their anticancer activities. In this research, a series of novel imidazolium salts substituted with lithocholic acid (LCA) and alkyl chains of various lengths (S1–S10) were evaluated against colon cancer cells. A significant reduction in the viability and metabolic activity was obtained in vitro for DLD-1 and HT-29 cell lines when treated with tested salts. The results showed that the activities of tested agents are directly related to the alkyl chain length, where S6–S8 compounds were the most cytotoxic against the DLD-1 line and S4–S10 against HT-29. The research performed on the xenograft model of mice demonstrated a lower tendency of tumor growth in the group receiving compound S6, compared with the group receiving 5-fluorouracil (5-FU). Obtained results indicate the activity of S6 in the induction of apoptosis and necrosis in induced colorectal cancer. LCA-based imidazolium salts may be candidates for chemotherapeutic agents against colorectal cancer.

Abstract 6061: Novel β-Apopicropodophyllin derivative, JNC-1043, exerts anti-cancer and radiosensitizing effects

Abstract In this study, we reported novel anti-cancer drug candidate, JNC-1043{(5S)-5-(4-Benzyloxy-3,5-dimethoxy-phenyl)-5,9-dihydro-8H-furo[3',4':6,7] naphtho[2,3-d][1,3]dioxol-6-one}, synthesized from β-Apopicropodophyllin. Firstly, we analyzed anti-cancer effect of JNC-1043. We performed MTT assay to detect IC50 values on several cancer cells - breast, lung, pancreatic, blood, gastric, colorectal cancers. In these experiments, 0.1~1 μM IC50 values of JNC-1043 were calculated, and JNC-1043 treatments induced activation of caspase-3 and -8. Additionally, effective values of pharmacokinetic parameters in male rats and 2 weeks Dose Rate Finding (DRF) of JNC-1043 also detected. This result showed JNC-1043 is a promising anti-cancer candidate. Next, we tested whether radiosensitizing effect of JNC-1043. Each IC50 values of DLD-1 and HCT116 cell lines were calculated as 0.157, 0.1145, respectively. Cell counting assays were performed and then showed combination of JNC-1043 and γ-ionizing radiation (IR) enhanced cell death. JNC-1043 only- or IR only-treatment induced cell death by 50~60%, respectively, while combination of JNC-1043 and IR increased more 20~30% cell death than those of JNC-1043 only- or IR only-treatment in both cell lines. Clonogenic assay also performed, and then dose enhancement ratios (DER) of DLD-1 and HCT116 were calculated as 1.40, 1.53, respectively. Apoptosis also was detected with Annexin V-Propidium iodide assay. Combination of JNC-1043 and IR increased apoptosis more 30~40% than those of JNC-1043 only- or IR only-treatment. DCFDA-based assay also were performed to indicate ROS production was enhanced in combination of JNC-1043 and IR. In vivo xenograft experiment, combination of JNC-1043 and IR reduced tumor sizes less 50% and increased apoptosis more 30~40% than those of JNC-1043 only- or IR only-treatment. These results implied anti-cancer and radiosensitizing effect of JNC-1043 resulted from promotion of apoptosis via enhancement of ROS production. Citation Format: Jong Kuk Park, A-Ram Kang, Jin-Hee Kwon, Jong-Ryoo Choi, Jie-Young Song, Sang-Gu Hwang. Novel β-Apopicropodophyllin derivative, JNC-1043, exerts anti-cancer and radiosensitizing effects [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 6061.

Abstract 5987: Membrane spanning 4 domains A12 (MS4A12) c.C211T stop-gain single-nucleotide polymorphism (rs2298553) promotes metastasis in colorectal cancer via the Hippo pathway

Abstract Background: Colorectal cancer (CRC) is one of the most malignant tumors worldwide. The prognosis of patients with colorectal cancer remains poor, with a 5-year metastasis survival rate of 20%-30%. Membrane spanning 4 domains subfamily A (MS4A) is an evolving family of structurally related cell surface proteins. Previous studies demonstrated that MS4A12 expression in CRC was significantly higher than that in the normal tissues. And low expression of MS4A12 in CRC was associated with poor survival in patients. However, the impact and mechanism of MS4A12 c.C211T stop-gain single-nucleotide polymorphism (SNP) (rs2298553) on the metastatic phenotype of CRC have not previously been examined. Current opinion holds that SNP introducing premature termination codons could alter the stability and function of transcripts and proteins. The aim of this study was to investigate the correlation between MS4A12 stop-gain SNP and invasion and migration of CRC cells. Methods: Whole exome sequencing and target sequencing were used to screen SNP related to distant metastasis. Immunohistochemical staining of tissue microarray was used to detect proteins in CRC tumor cancer and normal tissues. Recombinant plasmids carrying wild-type (CC) and MS4A12 c.C211T stop-gain SNP (TT) were constructed and then transfected to colon cancer cell lines DLD1, RKO. Knockdown of MS4A12 used the short hairpin RNA (shRNA). Western blot was performed to detect the level of protein expression. Wound healing essay, migration and invasion assays were used to detect metastatic ability of CRC. RNA-seq analysis, co-immunoprecipitation and mass assays were performed to further mechanism. Results: MS4A12 c.C211T stop-gain SNP (rs2298553) was screened and identified to be associated with CRC metastasis. Tissue microarray revealed that the expression of MS4A12 was lower in the colorectal tumor tissues compared to normal tissues. The low expression of MS4A12 in CRC was associated with a poor prognosis in patients. The expression level of the TT group was close to that of the basal level of empty vector transfected groups, which suggested that the c.C211T stop-gain SNP (rs2298553) inhibited truncated MS4A12 protein expression. Compared to the CC group, the TT group showed significantly increased invasion and migration ability. Meanwhile, MS4A12 knockdown also promoted invasion and migration. GO and KEGG pathway analysis suggested that the Hippo signaling pathway might be associated with the inhibition of migration and invasion by MS4A12. Compared to the CC group, the TT group impedes YAP/TAZ phosphorylation and facilitates YAP/TAZ nuclear translocation. Conclusion: MS4A12 c.C211T stop-gain SNP (rs2298553) promotes the invasion and migration of CRC. These results provide an important theoretical basis for studying the invasion and migration of CRC. Citation Format: Shuang Liu, Ting Jiang, Yan Yuan, Haina Yu, Zhiwei Guo, Yuanhong Gao, Yan Li, Weiwei Xiao. Membrane spanning 4 domains A12 (MS4A12) c.C211T stop-gain single-nucleotide polymorphism (rs2298553) promotes metastasis in colorectal cancer via the Hippo pathway [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 5987.

Noncanonical NF-κB factor p100/p52 regulateshomologous recombination and modulates sensitivity to DNA-damaging therapy.

Homologous recombination (HR) serves multiple roles in DNA repair that are essential for maintaining genomic stability, including double-strand DNA break (DSB) repair. The central HR protein, RAD51, is frequently overexpressed in human malignancies, thereby elevating HR proficiency and promoting resistance to DNA-damaging therapies. Here, we find that the non-canonical NF-κB factors p100/52, but not RelB, control the expression of RAD51 in various human cancer subtypes. While p100/p52 depletion inhibits HR function in human tumor cells, it does not significantly influence the proficiency of non-homologous end joining, the other key mechanism of DSB repair. Clonogenic survival assays were performed using a pair DLD-1 cell lines that differ only in their expression of the key HR protein BRCA2. Targeted silencing of p100/p52 sensitizes the HR-competent cells to camptothecin, while sensitization is absent in HR-deficient control cells. These results suggest that p100/p52-dependent signaling specifically controls HR activity in cancer cells. Since non-canonical NF-κB signaling is known to be activated after various forms of genomic crisis, compensatory HR upregulation may represent a natural consequence of DNA damage. We propose that p100/p52-dependent signaling represents a promising oncologic target in combination with DNA-damaging treatments.

Combined experimental and theoretical analyses on design, synthesis, characterization, and in vitro cytotoxic activity evaluation of some novel imino derivatives containing pyrazolone ring

In this research, some novel Schiff base derivatives 10–18 were synthesized for the first time, characterized, and tested for their anticancer activities. Spectroscopic characterization of the synthesized molecules 1–18 was carried out by using elemental analysis (C, H, N, S), FT-IR, HRMS, 1H- and 13C- NMR and DEPT-135 spectroscopic techniques. The antiproliferative activity studies of all newly synthesized compounds were tested against different human cancer cell lines, including colon and liver, using an MTT assay for 48 h. Under specified experimental conditions, three molecules (13-15) demonstrated higher cytotoxic activity than other molecules (1-12, 16-18) toward human epithelial colon colorectal cancer cell line (DLD-1) with IC50 values of 67.88, 56.53, and 48.70 μM, respectively. Besides, two different molecules (8 and 17) were found to have more toxic effects than molecules (1-7, 9-16, 18) against human liver epithelial hepatocellular carcinoma cell line (HepG2) with IC50 values of 65.72 and 54.84 μM, respectively. The selectivity of these compounds, which show high activity, was also tested on a healthy human cell line (Wl-38). Compounds (1–18) were docked against target proteins (PDB ID: 5ETY and 6V9C) representing DLD-1 and HepG2 cell lines.

GSTs, MRP and Apoptototic Markers in DLD-1 Human Colon Cancer Cell Line Before and After 5-FU Treatment

Multidrugresistance is an important factor limiting the effect of chemotherapy on cancer treatment. Disorders of drug transport and apoptosis, deterioration of redox homeostasis are among the main mechanisms that lead to multidrug resistance. The aim of this study was to determine the effect of 5-FU on GST isozymes, drug resistance proteins and apoptotic proteins before and after 5-Flourouracil application on DLD-1 colon cancer cell line. The cytotoxic effect of 5-FU was measured by WST-1test and, the efficiency of drug application was, also, proved by double staining via Hoechst 33342 with Propidium iodide. Next, the expression levels of GST isozymes, drug resistance proteins and apoptotic proteins were determined by immunocytochemistry. The cytotoxic effect of 5-FU at different doses on DLD-1 colon cancer cell line was determined by WST-1 method. MRP-2, 3, 6, 7 of drug resistance proteins; GSTA1, GSTM1, GSTT1, GSTZ1, GSTK1 and GSTO1 of GST proteins; bcl-2, caspase-3, p38, and p53, which are apoptotic proteins, have higher expression in the drug-treated DLD-1 cell line. GSTS1, MDR-1 and MRP-1expressions were not immunocytochemically different. It was determined that there is a direct correlation between the level of cytotoxicity and applied drug concentration. The cytotoxic effect of the drug increased with the increase in the dose of the drug. In this study, as first in the literature, the expression levels of some apoptotic markers, GST isozymes and drug resistance proteinswere evaluated togetherand except GSTS1, MDR-1 and MRP-1, they were all upregulated with respect to the control group after 5-FU administration.

Flavonoid Glycoside Diosmin Induces Apoptosis and Cell Cycle Arrest in DLD-1 Human Colon Cancer Cell Line

The purpose of this study was to see if diosmin induced apoptosis and inhibited cell proliferation in DLD-1 cell lines. The assays such as MTT, DCFDA, AO/EtBr, cell cycle analysis, annexin V-FITC apoptosis by flow cytometry, gene expression analysis were done to evaluate the in vitro anti-colon cancer potential of diosmin in DLD-1 human colon cancer cell line. While analyzing the results of MTT assay it is observed that when different concentrations of diosmin were induced, it was discovered that DLD-1 cells were killed in a dose-dependent manner. Diosmin mediated apoptosis induction is observed through AO/EtBr double staining method. Diosmin treatment in DLD-1 cell line generates high amount of ROS which is being analyzed using ROS DCFDA assay. While using flow cytometry to investigate the effect of diosmin on cell cycle distribution, it was found that diosmin-induced cell cycle arrest occurs in the S phase. When diosmin is administered, the proportion of surviving cells decreases, confirming the existence of cell death induction via apoptosis using the annexin V staining method. Diosmin upregulated mRNA levels expression of p53 and p21, indicating that diosmin induced S phase arrest and apoptosis in DLD-1 cells is controlled in part by the p21/p53 signaling pathway.

Curcumin Nanoparticles as Promising Anticancer Agent Against Human Colorectal Cancer Cell Lines

Curcumin, the main bioactive material found in turmeric, has been extensively studied as a curative measure for variety of diseases. Besides its high therapeutic value, curcumin possess some inherent limitations like poor aqueous solubility, less bioavailability, rapid metabolic rate, and fast elimination from the body. In the present study, nanoformulation of curcumin was discussed to improve its potentials and to reduce the disadvantages. The curcumin nanoparticles have been synthesized by wet-milling technique and have been characterized using FTIR, UV-Visible spectroscopy, SEM-EDS, TEM, and XRD. The results showed that the particles have a uniform dispersion and possess spherical shape with size ranging from 10-30 nm. FTIR studies showed that there was no change in the chemical structure of curcumin during the nanoparticle synthesis process. The anticancer activity of the synthesized nanoparticles was studied against DLD-1 (colorectal cancer) cell lines and cytotoxic evaluation was done against L929 (fibroblast) cells. It was observed that the curcumin nanoparticles significantly reduce the cell viability of DLD-1 cells in a concentration dependent manner and less toxic on normal L929 cells.

Cytotoxicity Evaluation and Antioxidant Activity of a Novel Drink Based on Roasted Avocado Seed Powder.

The avocado seed is an underused waste resulting from the processing of pulp. Polyphenols, fibers, and carotenoids are present in the seed, which also exhibits prophylactic, fungicidal, and larvicidal effects. Developing food products with avocado seed as an ingredient or spice is highly desired for nutritional, environmental, and economic reasons. The present study proposed its valorization in a hot drink, similar to already existing coffee alternatives, obtained by infusing the roasted and grinded avocado seed. The proximate composition of the raw or conditioned avocado seed and that of the novel drink were determined. The total phenolic content was assessed using the Folin-Ciocâlteu method. The total carotenoids were extracted and assessed spectrophotometrically. Starch determination was performed by the Ewers Polarimetric method. The highest content of polyphenols, 772.90 mg GAE/100 g, was determined in the crude seed, while in the drink was as low as 17.55 mg GAE/100 g. However, the proposed drink demonstrated high antioxidant capacity, evaluated through the DPPH method. This might be due to the high content of the total carotenoid compounds determined in the roasted seed (6534.48 µg/100 g). The proposed drink demonstrated high antiproliferative activity on Hs27 and DLD-1 cell lines.

Camptothecin, triptolide, and apoptosis inducer kit have differential effects on mitochondria in colorectal carcinoma cells.

Mitochondrial fission and fusion are required for cell survival, and several studies have shown an imbalance between fission and fusion in cancer. High levels of mitochondrial fusion are observed in drug‐resistant tumor cells, whereas mitochondrial fission may be important in sensitizing tumor cells to chemotherapy drugs. Based on current knowledge, we hypothesized that different chemotherapeutics might differentially affect mitochondrial dynamics and energy production. Thus, we selected chemotherapeutics with different mechanisms of action (camptothecin, triptolide and apoptosis inducer kit) and investigated their effect on mitochondria in colorectal carcinoma cells. We report that these chemotherapeutics decreased the activity of complex I and reduced the mitochondrial membrane potential, and also decreased the size of mitochondria in the colorectal carcinoma cell lines DLD1 and HCT‐116. Treatment with camptothecin, triptolide and/or apoptosis inducer kit results in differential effects of fission on apoptosis in these cells. Our results suggest that fission is an important process in apoptosis induced by chemotherapeutics.

Explorations of ATP-Binding Cassette Transporters and Apoptosis Signal Pathways of 2-Hydroxyanthraquinone Substituted Cyclotriphosphazenes in MCF-7 and DLD-1 Cell Lines.

As a class with biological properties, such as anti-cancer, anti-bacterial, anti-HIV, and various physical effects, phosphazene derivatives constitute the most striking part of inorganic compounds. Anthraquinones, on the other hand, are a broad family of compounds with a wide variety of biological properties; the biologically active anthraquinones have been used as valuable compounds for biochemical and pharmacological research. In this study, we aimed to investigate the effect of the anthraquinone substituted cyclotriphosphazene compounds on apoptosis and drug resistance in MCF-7 and DLD-1 cells. In breast and colon cells, mRNA levels of multi-drug resistance genes (ABCB1, ABCC3, ABCC10, ABCC11, and ABCG2), apoptotic genes (BAX, BCL-2, p53, and PARP), heat shock (HSP27, HSP40, HSP60, HSP90α) and endoplasmic reticulum chaperone genes (GRP78, and GRP94) were determined by the qPCR method. The amount of proteins of the cell cycle, HSPs, apoptosis, and related signaling pathways were measured by the membrane array kits. Compounds 2, 3, 4, and 7 showed the most potent results on the ATP-binding cassette genes in both breast and colon cancer cells. These compounds have a remarkable effect on apoptotic, heat shock, and ER chaperone genes in cancer cells. Besides, these compounds induced protein levels of pro-apoptotic pathways, leading to apoptosis by inhibiting anti-apoptotic pathways. Also, these compounds decreased HSPs. These compounds have potential properties that eliminate drug resistance, suppress heat shock and ER chaperone genes, and drag cells to apoptotic cell death and are notable for drug studies.

Cembranolides and Related Constituents from the Soft Coral Sarcophyton cinereum.

In an attempt to explore the bioactive metabolites of the soft coral Sarcophyton cinereum, three new cembranolides, cinerenolides A–C (1–3), and 16 known compounds were isolated and identified from the EtOAc extract. The structures of the new cembranolides were elucidated on the basis of spectroscopic analysis, and the NOE analysis of cinerenolide A (1) was performed with the assistance of the calculated lowest-energy molecular model. The relative configuration of cinerenolide C (3) was determined by the quantum chemical NMR calculation, followed by applying DP4+ analysis. In addition, the cytotoxic assays disclosed that some compounds exhibited moderate to potent activities in the proliferation of P388, DLD-1, HuCCT-1, and CCD966SK cell lines.

The influence of the pH on the incorporation of caffeic acid into biomimetic membranes and cancer cells

Caffeic acid (CA) is a phenolic compound synthesized by all plant species. It constitutes the main hydroxycinnamic acid found in human diet and presents a variety of beneficial effects including anticancer activity. Current data suggests essential role of the interplay between anticancer drugs and the cell membrane. Given this, biophysical interactions between CA and cancer cells or biomimetic membranes were investigated. Glioblastoma cell line U118MG and colorectal adenocarcinoma cell line DLD-1, as well as lipid bilayers and liposomes, were used as in vitro models. Electrophoretic light scattering was used to assess the effect of CA on the surface charge of cancer cells and liposomal membranes. Electrochemical impedance spectroscopy was chosen to evaluate CA-dependent modulatory effect on the electrical capacitance and electrical resistance of the bilayers. Our results suggest that CA fulfills physicochemical criteria determining drug-like properties of chemical compounds, and may serve as a potential cytostatic agent in cancer treatment.

The Interaction Between Cancer-associated Fibroblasts and Cancer Cells Enhances Bcl-xL and Mcl-1 in Colorectal Cancer.

The acquisition of resistance to apoptosis is one of the biggest problems in colorectal cancer (CRC) treatment. This study aimed to elucidate the mechanisms of resistance to apoptosis with a focus on interleukin (IL)-6 produced by the interaction between cancer cells and cancer-associated fibroblasts (CAFs). DLD-1 and HCT116 cell lines were treated with IL-6 and furthermore co-cultured with CAFs. The expression levels of Bcl-xL, Mcl-1 and phosphorylation of STAT3 were evaluated by western blotting. We also performed immunostaining for CRC specimens and evaluated the correlation between CAFs invasion and Bcl-xL/Mcl-1 expression. Both IL-6 and co-culturing enhanced Bcl-xL, Mcl-1 and the phosphorylation of STAT3. Immunohistochemistry showed a positive correlation between CAFs and Bcl-xL/Mcl-1. These results showed that the interaction between CAFs and cancer cells enhances Bcl-xL and Mcl-1 through the IL-6/STAT3 signaling pathway. Our findings provide new potential therapeutic targets and strategies for CRC treatment.

Synthesis and characterization of silver nanoparticles using curcumin: cytotoxic, apoptotic, and necrotic effects on various cell lines.

Nanostructures have distinctive chemical and physical features owing to their surface area and nanoscale size. In this study, silver nanoparticles were synthesized using curcumin, a medicinally valuable natural product. The structure of curcumin-mediated silver nanoparticles (c-AgNPs) was identified by extensive spectroscopic techniques. The maximum absorption was observed at 430nm in UV-Vis spectrum. The crystal structure of c-AgNPs was identified by XRD. The morphology of the structure was determined by SEM image. The particle size was found as 51.13nm. The functional groups of curcumin and c-AgNPs were established by FTIR spectroscopy. Cytotoxic activity of c-AgNPs was carried out using A549, DLD-1, and L929 with MTT assay. c-AgNPs revealed excellent activity on DLD-1cell lines and A549cell lines at 1.0mg/mL concentration with the lethal effect of 80%. However, nanoparticles did not show the considerable effect on L929. Moreover, they induced apoptosis. Consequently, c-AgNPs are a promising material for anticancer drugs candidate.