Colon Cancer Cell Death Research Articles

Silver nanoparticles induce cell death of colon cancer cells through impairing cytoskeleton and membrane nanostructure

Globally, colon cancer is a predominant cause of increased morbidity and mortality annually; therefore, in addition to traditional treatments, new protocols are under continuous investigation. Nanotechnology-based cancer therapy is a new strategy and considered one of the most promising research directions for colon cancer. In this study, we used a silver nanoparticle (AgNP)-based methodology to treat colon cancer cells, and single cell approaches to examine how AgNPs exerted inhibiting effects on cells. We found that AgNPs could apparently destroy cytoskeleton and topography structures, alter cell membrane nanostructures, and thus increase membrane roughness, and depress cell membrane adhesion properties and cell stiffness. We also found that AgNPs caused mitochondrial dysfunctions including hyperpolarization of membrane potential and reactive oxygen species (ROS) accumulation. Notably, AgNPs altered all phenotypes or functions of cells in a dose-dependent manner. Therefore, our research provided a new paradigm for revealing killing mechanisms of AgNPs against colon cancer cells from single cell biophysical aspects, which could advance AgNP-based nanotechnology cancer therapy.

Hypotonic stress enhances colon cancer cell death induced by platinum derivatives and immunologically improves antitumor efficacy of intraperitoneal chemotherapy.

Colorectal cancer is a highly metastatic disease that could invade various distal organs and also the peritoneal cavity leading to peritoneal carcinomatosis. This is a terminal condition with poor prognosis and only palliative treatments such as cytoreductive surgery and intraperitoneal chemotherapy are proposed to some patients. However, clinicians use different parameters of treatments without any consensus. Here we decided to evaluate the effect of osmolarity in the efficacy of this procedure to kill colon cancer cells. We first show that a short exposure of platinum derivatives in hypotonic conditions is more efficient to decrease cell viability of human and murine colon cancer cells in vitro as compared to isotonic conditions. This is related to more important incorporation of platinum and the capacity of hypotonic stress to induce the copper transporter CTR1 oligomerization. Oxaliplatin in hypotonic conditions induces caspase-dependent cell death of colon cancer cells. Moreover, hypotonic conditions also modulate the capacity of oxaliplatin and cisplatin (but not carboplatin) to induce immunogenic cell death (ICD). In vivo, oxaliplatin in hypotonic conditions increases CD8+ T cell tumor infiltration and activation. Finally, in a murine peritoneal carcinomatosis model, oxaliplatin in hypotonic conditions is the only tested protocol which is able to slow down the appearance of tumor nodules and increase mice survival, while showing no effect in CD8+ T cells depleted mice or in immunodeficient mice. Altogether, our study provides new information both in vitro and in a preclinical model of peritoneal carcinomatosis, which highlights the importance of hypoosmolarity in intraperitoneal chemotherapy.

MYC promotes tryptophan uptake and metabolism by the kynurenine pathway in colon cancer.

Tumors display increased uptake and processing of nutrients to fulfill the demands of rapidly proliferating cancer cells. Seminal studies have shown that the proto-oncogene MYC promotes metabolic reprogramming by altering glutamine uptake and metabolism in cancer cells. How MYC regulates the metabolism of other amino acids in cancer is not fully understood. Using high-performance liquid chromatography (HPLC)-tandem mass spectrometry (LC-MS/MS), we found that MYC increased intracellular levels of tryptophan and tryptophan metabolites in the kynurenine pathway. MYC induced the expression of the tryptophan transporters SLC7A5 and SLC1A5 and the enzyme arylformamidase (AFMID), involved in the conversion of tryptophan into kynurenine. SLC7A5, SLC1A5, and AFMID were elevated in colon cancer cells and tissues, and kynurenine was significantly greater in tumor samples than in the respective adjacent normal tissue from patients with colon cancer. Compared with normal human colonic epithelial cells, colon cancer cells were more sensitive to the depletion of tryptophan. Blocking enzymes in the kynurenine pathway caused preferential death of established colon cancer cells and transformed colonic organoids. We found that only kynurenine and no other tryptophan metabolite promotes the nuclear translocation of the transcription factor aryl hydrocarbon receptor (AHR). Blocking the interaction between AHR and kynurenine with CH223191 reduced the proliferation of colon cancer cells. Therefore, we propose that limiting cellular kynurenine or its downstream targets could present a new strategy to reduce the proliferation of MYC-dependent cancer cells.

NO2 functionalized coumarin derivatives suppress cancer progression and facilitate apoptotic cell death in KRAS mutant colon cancer

Colon cancer is one of the most lethal cancers worldwide even with the significant progress made in screening techniques and therapeutic agents. Genetic mutations in tumors complicated the treatments, and the survival rate remains low for patients at late or metastatic stages. KRAS gene mutation which leads to failure of the EGFR targeted therapies stands for an example of the challenges in clinical sites. Therefore, development of novel agents for colon cancer treatment is in need. Natural and synthetic coumarin derivatives have been suggested with various biological activities with pharmacologic potential including anti-cancer capacity. Here in this study, five coumarin derivatives, include trifluoromethyl-, dimethoxy-, and/or nitro-substitutions at different positions, were synthesized. Their cancer inhibition potential was investigated in various cancer cell lines. Our data demonstrated that one nitro-coumarin derivate, 5,7-Dimethoxy-4-methyl-6-nitro-chromen-2-one, exhibits cytotoxicity specifically towards colon cancer cells under competitive EC50. Our results showed that this compound can effectively suppress colon cancer cells harboring either wild type or mutant KRAS genes, and that it could inhibit short-term proliferation, long term proliferation, and migration capacities of cancer cells. Finally, we demonstrated that this coumarin derivate facilitates cancer cell death through activation of apoptosis pathway. Our results suggest that this coumarin derivate is a promising lead drug worth further investigation and development for future cancer treatment.

Naringenin-loaded nano-structured lipid carrier fortifies oxaliplatin-dependent apoptosis in HT-29 cell line

Abstract In the present study, we inspected the role of naringenin-loaded nanostructured lipid carriers (NLCs) in the induction of apoptosis in (HT-29) colon cancer cell in co-administration with oxaliplatin. The cytotoxicity behavior of nanoparticles was investigated by MTT assay and DAPI staining. To understand the molecular mechanism of formulation in induction program cell death, flow cytometric and Real-Time RT-PCR assays were applied. New formulation revealed a mean particle size of 98 ± 3 nm with homogenous dispersity, which was confirmed by SEM graphs. Naringenin and Oxaliplatin-induced cell death in HT-29 colon cancer by IC50 value of 150 ± 11.04 μM and 20 ± 2.25 μM, respectively. Loading Naringenin into new formulation declined the proportion of cell proliferation of HT-29 colon cancer from 47 ± 9.1% to 27 ± 5.3% (p

Preparing Sodium Alginate/Polyethyleneimine Spheres for Potential Application of Killing Tumor Cells by Reducing the Concentration of Copper Ions in the Lesions of Colon Cancer.

Inhibition of residual malignant tumors in patients with colon cancer after operation is one of the difficulties in rehabilitation treatment. At present, using biocompatible materials to remove the copper ion which is the growth dependence of malignant tumors in the lesion site is considered to be the frontier means to solve this problem. In this work, we developed a sodium alginate (SA)/polyethyleneimine (PEI) hydrogel sphere via cross-linking method (SA/SP/SA; SP = SA/PEI) as an oral biomaterial for adsorbing and removing copper ions from colon cancer lesions. The evaluated results showed that the SA/PEI/SA (SPS) hydrogel sphere obtained the largest swelling rate at pH 8.3 which was the acid-base value of colon microenvironment and absorbed more copper ions compared with the SA control. The cell experiment presented that the SPS hydrogel sphere owned better compatibility on normal fibroblasts and promoted higher death of colon cancer cells compared with SA/PEI (SP) and SA control. Our data suggested that the SA/PEI hydrogel sphere had the potentiality as an oral biomaterial for inhibiting colon cancer cells.

Synthetic derivatives of the natural product 13-amino 2-desoxy-4-epi-pulchellin inhibit STAT3 signaling and induce G2/M arrest and death of colon cancer cells

2-Desoxy-4-epi-puchellin (PCL) is a sesquiterpene-lactone, which naturally occurs in many traditional Chinese medicinal plants, and has antitumor and anti-inflammatory activities. In this work, a series of 13-amino derivatives of PCL were synthesized through Michael addition reaction. Inhibition of IL-6-induced STAT3 signaling pathway and in vitro cytotoxicity of these compounds were evaluated, and their effect on the cell cycle was also studied. The methyl hydroxyethylamine derivatives showed higher potency than PCL, which could induce significant mitotic arrest via G2/M arrest in HCT116 cancer cells, indicating that these derivatives have the potential to be developed into anti-colon cancer drugs.

The Long-lasting Radioprotective Effect of Caffeic Acid in Mice Exposed to Total Body Irradiation by Modulating Reactive Oxygen Species Generation and Hematopoietic Stem Cell Senescence-Accompanied Long-term Residual Bone Marrow Injury.

Total body irradiation (TBI) serves as an effectively curative therapy for cancer patients and adversely causes long-term residual bone marrow (BM) injury with premature senescence of hematopoietic stem cells (HSCs), which is mediated by increased production of reactive oxygen species (ROS). In the present study, we investigated how the exposure time of TBI in a mouse model affects HSCs and whether the treatment of caffeic acid (CA), a known dietary phenolic antioxidant, has a radioprotective effect. Single (S)-TBI at a sublethal dose (5 Gy) caused relatively higher induction of mitochondrial ROS and senescence-related factors in HSCs than those in hematopoietic progenitor cells (HPCs) and Lineage-Sca-1+c-Kit+ (LSK) cells, as well as reduced clonogenic formation and donor cell-derived reconstituting capacity. Repetitive double (D)-TBI (two months after the S-TBI at a dose of 5Gy) further weakened HSPC function via mitochondrial ROS accumulation and senescence-associated β-galactosidase (SA-β-gal) activity. Oral administration of CA (20 mg/kg) five times before and once immediately after TBI ameliorated ROS generation and TBI-induced HSC senescence and its radioprotective effect was long lasting in S-TBI mice but not in D-TBI mice. Further, supplementation of CA also induced apoptotic cell death of colon cancer cells. Collectively, these findings indicate that CA has a dual effect, ameliorating HSC senescence-accompanied long-term BM injury in S-TBI mice and stimulating apoptotic cell death of colon cancer cells.

Investigation of Anticancer Activity of Lindernia crustacea (L.) F. Muell. var. Crustacean

Lindernia crustacea (L.) F. Muell. var. crustacean or “Ya Kap Hoi: YKH” was misunderstood as “Ya Yad Nam Kang” and cancer curative. This study aimed to investigate anticancer activity by the analysis of chemical constituent in extracts and in vitro screening of biological activities of the extract in several aspects such as cancer cell lines cytotoxic activity, immune cell proliferating activity, reducing power activity, alkylation activity. The HPLC analysis showed the absence of plumbagin peak observed in the HPLC chromatograms of both YKH aqueous and YKH ethanolic extracts. The YKH ethanolic extract yielded more chemical constituents than that of the YKH aqueous extract. The YKH aqueous extract was inactive against all cancer cells tested. Interestingly, YKH ethanolic extract caused cancer cell death in HCT116 colon cancer, HepG2 liver cancer, and Jurkat leukemic cancer cell lines in the concentration dependent manner. The following IC50 concentrations of the YKH ethanolic extract that possessed 50 % cell death after 24 h exposure in HCT116, HepG2, and Jurkat cell line were 195.4 ± 12, 171.7 ± 8.7, and 48.8 ± 5.7 µg/mL, respectively. Both aqueous and ethanolic extracts of YKH showed high antioxidant activity based on reducing power activity but did not have alkylation activity. At high concentration (250 µg/mL), YKH ethanolic extract can inhibit immune cells proliferation activity more than the YKH aqueous extract. An unexpected but critical outcome of our studies was the finding that anticancer activity is promised by selecting the plant extraction solvent with less polarity. Potential anticancer constituents were extracted from YKH using ethanol and these constituents cannot be found in aqueous solution of YKH.

Sequential treatment with celecoxib and bortezomib enhances the ER stress-mediated autophagy-associated cell death of colon cancer cells.

Treatment with celecoxib and bortezomib as single chemotherapeutic agents reduces the viability and proliferation of colorectal cancer cells. The use of these agents in combination with other chemotherapeutic agents is usually associated with adverse effects. In the present study, a combination of celecoxib and bortezomib was investigated for potential synergistic effects in colon cancer cells. The sequential exposure to celecoxib with bortezomib synergistically induced apoptotic death in human colon cancer cells compared with groups treated with a single drug or other drug combinations. c-Jun N-terminal kinase/p38-mitogen-activated protein kinase-induced endoplasmic reticulum (ER) stress through serial exposure to celecoxib and bortezomib may have induced the intracellular Ca2+ release, leading to the generation of autophagosomes in p53-expressing HCT-116 cells. Targeted inhibition of p53 activity or ER stress or treatment with the Ca2+-chelating agent BAPTA-AM suppressed the ER stress-mediated Ca2+ release and apoptosis. Although p53−/− HCT-116 cells were less sensitive to sequential treatment with celecoxib and bortezomib, co-localization of autophagosomes was detected in the absence of CCAAT-enhancer-binding protein homologous protein expression. Treatment of p53−/− HCT-116 cells with BAPTA-AM did not inhibit apoptosis following serial treatment with celecoxib and bortezomib. These results suggest that the order of drug administration is important in treating cancer and that the sequential treatment with celecoxib and bortezomib enhances the ER stress-mediated autophagy-associated cell death of colon cancer cells, regardless of p53 expression.

Abstract 2675: Quinacrine-mediated activation of JNK1 inhibits Nrf2 and causes cell death in human colon cancer

Abstract Up-regulation of Nrf2 is associated with poor prognosis in skin, breast, prostate, lung, head and neck, endometrial, and colorectal cancer, and increases chemoresistance and radioresistance by inducing antioxidant and detoxification genes. Nrf2 inhibitors are not currently available for clinical use. Here, we investigated the feasibility of using quinacrine, an old antimalarial drug, as a novel Nrf2 inhibitor for treating colorectal cancer. Immunohistochemical analysis of colorectal cancer samples from 60 patients was used to assess the correlation between Nrf2 expression levels and clinicopathological features. Using data from tumors in the Cancer Genome Atlas (n=290 normal, n=295 cancer) and publicly available data sets, we validated the correlation between Nrf2 expression and poor prognosis in colorectal cancer. The effect of quinacrine on the activation of JNK1-mediated Nrf2 degradation was analyzed with co-immunoprecipitation, immunoblotting, and luciferase reporter assays. To confirm that quinacrine-mediated Nrf2 degradation caused cancer cell death, we used Nrf2 gain-of-function and loss-of-function experiments. The cytotoxicity of quinacrine against colorectal cancer cells under both normoxia (20% O2) and hypoxia (0.5% O2) was assessed by clonogenic survival assays. Finally, a xenograft mouse model was used to confirm the results of our in vitro experiments in an in vivo setting. High Nrf2 levels were correlated with increased expression of its target genes and poor survival among colorectal cancer patients in immunohistochemical analysis and analysis for publicly available data sets. The quinacrine-mediated activation of JNK1 induced Nrf2 degradation by increasing the interaction between Keap1 and Nrf2, thereby causing cancer cell death under normoxia and hypoxia and suppressing tumor growth in our in vivo model. Overexpression of Nrf2 effectively prevented quinacrine-mediated Nrf2 degradation and cancer cell death regardless of oxygen tension. Additionally, siRNA-mediated JNK1 knockdown inhibited quinacrine-mediated Nrf2 degradation. Luciferase reporter assays and qPCR revealed that quinacrine inhibited Nrf2-mediated transcription in cancer cells under normoxia and hypoxia. Finally, the quinacrine-mediated degradation of Nrf2 was found to increase the sensitivity of colorectal cancer in in vivo model and cancer cells under both normoxia and hypoxia in vitro to ROS-inducing drugs and anticancer drugs commonly used against colorectal cancer. Quinacrine caused colorectal cancer cell death by promoting activation of JNK1-mediated Nrf2 degradation. Citation Format: Eun-Taex Oh, Chan-Woo Kim, Ha Gyeong Kim, In-Seon Lee, Chae Won Lee, Heon Joo Park. Quinacrine-mediated activation of JNK1 inhibits Nrf2 and causes cell death in human colon cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 2675.

1-L-MT, an IDO inhibitor, prevented colitis-associated cancer by inducing CDC20 inhibition-mediated mitotic death of colon cancer cells.

Indoleamine 2,3-dioxygenase 1 (IDO1), known as IDO, catabolizes tryptophan through kynurenine pathway, whose activity is correlated with impaired clinical outcome of colorectal cancer. Here we showed that 1-L-MT, a canonical IDO inhibitor, suppressed proliferation of human colorectal cancer cells through inducing mitotic death. Our results showed that inhibition of IDO decreased the transcription of CDC20, which resulted in G2/M cycle arrest of HCT-116 and HT-29. Furthermore, 1-L-MT induced mitochondria injuries and caused apoptotic cancer cells. Importantly, 1-L-MT protected mice from azoxymethane (AOM)/dextran sodium sulfate (DSS)-induced colon carcinogenesis, with reduced mortality, tumor number and size. What is more, IDO1-/- mice exhibited fewer tumor burdens and reduced proliferation in the neoplastic epithelium, while, 1-L-MT did not exhibit any further protective effects on IDO-/- mice, confirming the critical role of IDO and the protective effect of 1-L-MT-mediated IDO inhibition in CRC. Furthermore, 1-L-MT also alleviated CRC in Rag1-/- mice, demonstrating the modulatory effects of IDO independent of its role in modulating adaptive immunity. Taken together, our findings validated that the anti-proliferation effect of 1-L-MT in vitro and the prevention of CRC in vivo were through IDO-induced cell cycle disaster of colon cancer cells. Our results identified 1-L-MT as a promising candidate for the chemoprevention of CRC.

Silencing Livin improved the sensitivity of colon cancer cells to 5-fluorouracil by regulating crosstalk between apoptosis and autophagy.

Colorectal cancer (CRC) is the third most common cause of cancer-associated mortality worldwide. Currently, 5-fluorouracil (5-FU) remains a widely used chemotherapeutic drug in the treatment of CRC; however, 5-FU resistance during treatment has become a common problem. Livin, a member of the inhibitor of apoptosis protein family, is considered to be associated with tumor resistance to chemotherapy. In the present study, Livin-silenced cells were generated by introducing a lentivirus into HCT116 and SW620 colon cancer cell lines. Acridine orange/ethidium bromide staining was used as an indicator of cell death. Western blot analysis was performed to detect protein expression levels, and transmission electron microscopy was used to assess autophagy. The half-maximal inhibitory concentration of 5-FU in colon cancer cells was evaluated using a Cell Counting Kit-8 assay. The results of the present study confirmed that silencing Livin significantly enhanced colon cancer cell death in the presence of 5-FU, increased expression levels of various apoptosis- and autophagy-associated proteins and augmented chemotherapeutic sensitivity to 5-FU. Furthermore, the present study demonstrated that this effect may be reversed when autophagy or apoptosis was inhibited, indicating that apoptosis and autophagy were involved in this process. The protein kinase B signaling pathway and B-cell lymphoma-2 expression levels significantly decreased following Livin knockdown, suggesting they may contribute to the regulation of apoptosis and autophagy crosstalk, which caused the Livin knockdown-induced cell death observed.

Aldose reductase inhibitor, fidarestat prevents doxorubicin-induced endothelial cell death and dysfunction

Despite doxorubicin (Dox) being one of the most widely used chemotherapy agents for breast, blood and lung cancers, its use in colon cancer is limited due to increased drug resistance and severe cardiotoxic side effects that increase mortality associated with its use at high doses. Therefore, better adjuvant therapies are warranted to improve the chemotherapeutic efficacy and to decrease cardiotoxicity. We have recently shown that aldose reductase inhibitor, fidarestat, increases the Dox-induced colon cancer cell death and reduces cardiomyopathy. However, the efficacy of fidarestat in the prevention of Dox-induced endothelial dysfunction, a pathological event critical to cardiovascular complications, is not known. Here, we have examined the effect of fidarestat on Dox-induced endothelial cell toxicity and dysfunction in vitro and in vivo. Incubation of human umbilical vein endothelial cells (HUVECs) with Dox significantly increased the endothelial cell death, and pre-treatment of fidarestat prevented it. Further, fidarestat prevented the Dox-induced oxidative stress, formation of reactive oxygen species (ROS) and activation of Caspase-3 in HUVECs. Fidarestat also prevented Dox-induced monocyte adhesion to HUVECs and expression of ICAM-1 and VCAM-1. Fidarestat pre-treatment to HUVECs restored the Dox-induced decrease in the Nitric Oxide (NO)-levels and eNOS expression. Treatment of HUVECs with Dox caused a significant increase in the activation of NF-κB and expression of various inflammatory cytokines and chemokines which were prevented by fidarestat pre-treatment. Most importantly, fidarestat prevented the Dox-induced mouse cardiac cell hypertrophy and expression of eNOS, iNOS, and 3-Nitrotyrosine in the aorta tissues. Further, fidarestat blunted the Dox-induced expression of various inflammatory cytokines and chemokines in vivo. Thus, our results suggest that by preventing Dox-induced endothelial cytotoxicity and dysfunction, AR inhibitors could avert cardiotoxicity associated with anthracycline chemotherapy.

Yellow and green pigments from Calophyllum inophyllum L. seed oil induce cell death in colon and lung cancer cells.

Natural compounds have been candidates for anticancer medicine over the last 20 years. During the process of isolating seed oil from Calophyllum inophyllum L., yellow and green pigments containing multiple compounds with an aromatic structure were identified. High-performance liquid chromatography and nuclear magnetic resonance analysis of these pigments revealed that the compounds present were identical, but the concentration of the compounds was different. Treatment with the pigments was able to induce the death of DLD-1 human colon cancer cells and increase the percentage of the cells in the sub-G1 and sub-G2/M phases in a dose-dependent manner. Additionally, the pigments were able to exhibit cytotoxic activity on A549 and H1975 human non-small cell lung cancer (NSCLC) cell lines at 24 h, with half-maximal inhibitory concentrations (IC50) values of 0.1206 and 0.0676%, respectively for green pigments, and 0.0434 and 0.0501%, respectively for yellow pigments. Furthermore, a decrease in IC50 value was associated with an increase in the duration of treatment. However, a sharp decrease in IC50 value of the yellow pigment was observed for H1975 cells at 48 h and for A549 cells at 72 h compared with no change in IC50 value for the green pigment with time, suggesting that the pigments function and induce cell death differently in the two cell lines. An investigation was performed into the synergistic effect of the green pigment and gefitinib (Iressa®, ZD1839), which is a selective epidermal growth factor receptor-tyrosine kinase inhibitor to block growth factor-mediated cell proliferation. The combination of the green pigment and gefitinib resulted in an enhancement of the decrease in viability of A549 and H1975 cells compared with treatment with gefitinib alone, which suggested that treatment with the green pigments was able to enhance the sensitivity of NSCLC cells to gefitinib. In conclusion, these pigments may be considered for development as anti-colon cancer agents.

Ginkgolic acid induces interplay between apoptosis and autophagy regulated by ROS generation in colon cancer

Presently, developing effective anti-colon cancer drugs still remains to be important. Ginkgolic acids (GA), as a botanical drug extracted from the seed coat of Ginkgo biloba L., possess various bioactive properties. Our findings, for the first time, indicated that GA suppressed colon cancer cell proliferation, migration and invasion. GA led to cell death through G0/G1 phase arrest. In addition, apoptosis was significantly induced by GA treatment. The intrinsic apoptosis pathway was included, proved by the release of cytochrome c (Cyto-c) from the mitochondria into the cytosol. GA-induced autophagy was supported by the dose-dependent increase of LC3BII, autophagy-related gene-5 (ATG-5) and Beclin-1. Notably, silencing ATG-5 further reduced the cell viability and enhanced apoptosis in GA-treated colon cancer cells, indicating that GA-induced apoptosis rather than autophagy contributes to colon cancer cell death. And mammalian target of rapamycin complex 1 (mTORC1) was dose-dependently reduced by GA, evidenced by the reduction of p-mTOR, p-p70 ribosomal S6 kinase (p70s6k) and p-pras40. Moreover, GA markedly resulted in reactive oxygen species (ROS) generation, along with increased H2O2 and O2−. However, blocking ROS generation using its scavenger, NAC, significantly recovered GA-induced cells death, supported by the increase of cell viability, and the decrease of apoptosis. The expressions of autophagy- and cell cycle arrest-related molecules, as well as mTORC1 were also reversed by N-acetyl-l-cysteine (NAC) in GA-treated cells. In vivo, GA reduced tumor growth without toxicity to animals. In conclusion, our study illustrated that GA caused G0/G1 phase arrest and triggered intrinsic apoptosis and autophagy modulated by ROS generation in human colon cancer, elucidating that GA might be considered as a potential agent for colon cancer therapy.

Antitumor activity of HPA3P through RIPK3-dependent regulated necrotic cell death in colon cancer.

The antimicrobial peptide HPA3 shows anticancer activity in gastric cancer and leukaemia. However, how HPA3 exerts its anticancer activity, as well as whether it also exhibits activity in other cancers, remains unknown. Therefore, the aim of this study was to evaluate the anticancer activity of HPA3 and its analogues in colon cancer and to elucidate the mechanisms responsible for this activity. HPA3P decreased cell viability, whereas HPA3 and HPA3P2 did not decrease cell viability in colon cancer cells compared with control cells. This reduction in cell viability occurred through necrosis, a conclusion supported by our observation of the release of cellular contents, our intracellular PI staining results, and our observation of the release of HMGB1. Moreover, RIPK3 inhibition blocks the reduction of cell viability by HPA3P. Consistent with this finding, we found that knocking down RIPK3 and MLKL, key necroptosis proteins, attenuates the reductions in cell viability induced by HPA3P. Furthermore, HPA3P can improve the anticancer activity of chemotherapeutic agents and exhibits anticancer activity in other cancer cells. These results suggest that HPA3P may have potential as an anticancer agent in the treatment of colon cancer.

Curcumin induces apoptosis and cell cycle arrest via the activation of reactive oxygen species–independent mitochondrial apoptotic pathway in Smad4 and p53 mutated colon adenocarcinoma HT29 cells

Curcumin is a natural dietary polyphenol compound that has various pharmacological activities such as antiproliferative and cancer-preventive activities on tumor cells. Indeed, the role reactive oxygen species (ROS) generated by curcumin on cell death and cell proliferation inhibition in colon cancer is poorly understood. In the present study, we hypothesized that curcumin-induced ROS may promote apoptosis and cell cycle arrest in colon cancer. To test this hypothesis, the apoptosis-inducing potential and cell cycle inhibition effect of ROS induced by curcumin was investigated in Smd4 and p53 mutated HT-29 colon adenocarcinoma cells. We found that curcumin treatment significantly increased the level of ROS in HT-29 cells in a dose- and time-dependent manner. Furthermore, curcumin treatment markedly decreased the cell viability and proliferation potential of HT-29 cells in a dose- and time-dependent manner. Conversely, generation of ROS and inhibitory effect of curcumin on HT-29 cells were abrogated by N-acetylcysteine treatment. In addition, curcumin treatment did not show any cytotoxic effects on HT-29 cells. Furthermore, curcumin-induced ROS generation caused the DNA fragmentation, chromatin condensation, and cell nuclear shrinkage and significantly increased apoptotic cells in a dose- and time-dependent manner in HT-29 cells. However, pretreatment of N-acetylcysteine inhibited the apoptosis-triggering effect of curcumin-induced ROS in HT-29 cells. In addition, curcumin-induced ROS effectively mediated cell cycle inhibition in HT-29 cells. In conclusion, our data provide the first evidence that curcumin induces ROS independent apoptosis and cell cycle arrest in colon cancer cells that carry mutation on Smad4 and p53.

The Nedd8-activating enzyme inhibitor MLN4924 suppresses colon cancer cell growthviatriggering autophagy

Neddylation is a post-translational protein modification process. MLN4924 is a newly discovered pharmaceutical neddylation inhibitor that suppresses cancer growth with several cancer types. In our study, we first investigated the effect of MLN4924 on colon cancer cells (HCT116 and HT29). MLN4924 significantly inhibited the neddylation of cullin-1 and colon cancer cell growth in a time and dose-dependent manner. MLN4924 induced G2/M cell cycle arrest and apoptosis in HCT116 and HT29 cells. Moreover, MLN4924 also triggered autophagy in HCT116 and HT29 cells via suppressing the PI3K/AKT/mTOR pathway. Inhibiting autophagy by autophagy inhibitor 3-MA or ATG5 knockdown reversed the function of MLN4924 in suppressing colon cancer cell growth and cell death. Interestingly, MLN4924 suppresses colon cell growth in a xenograft model. Together, our finding revealed that blocking neddylation is an attractive colon cancer therapy strategy, and autophagy might act as a novel anti-cancer mechanism for the treatment of colon cancer by MLN4924.

Ultrasonic Modified Sweet Potato Pectin Induces Apoptosis like Cell Death in Colon Cancer (HT-29) Cell Line

ABSTRACTBackground: Pectin and especially modified citrus pectin possesses anticancer activity. Hence, the current study investigated anticancer activity of ultrasonic-modified sweet potato pectin (SPP) on HT-29 cells to assess its potential as a cancer therapeutic agent. Method: The effect of ultrasonic treatment on SPP molecular weight, galacturonic acid content, degree of methoxylation, and neutral sugar was investigated. Moreover, the effect of sonicated variant on human HT-29 cell proliferation was assessed by MTT assay, cell cytotoxicity, and apoptosis by Annexin V/PI flow cytometer and caspase-3 activity was studied. Results and discussion: Sonication led up to seven-fold decrease in molecular weight. The degree of methoxylation (DM) decreased more than two-fold. Moreover, the galacturonic acid (GalA) content increased up to 92%, arabinose and galactose content increased. The SSPP inhibited cell proliferation with the IC50 values 0.5 mg/ml and 0.75 mg/ml for 400 W and 200 W SSPP, respectively. Moreover, 14.41 ± 1.64% cell cytotoxicity was elicited by 400 W SSPP and 6.83 ± 0.80% by 200 W SSPP. Both SSPPs induced apoptosis with 400 W SSPP eliciting 19.42% and 42.21% apoptosis at 0.1 and 0.5 mg/ml, while 200 W SSPP induced 13.79% and 39.50% apoptosis at 0.1 and 0.5 mg/ml, respectively. SSPP activity increased with both increased concentration and sonication intensity.