Modulation of Colorectal Tumor Behavior via lncRNA TP53TG1-Lipidic Nanosystem

Long non-coding RNAs (lncRNAs) are an emerging group of RNAs with a crucial role in cancer pathogenesis. In gastrointestinal cancers, TP53 target 1 (TP53TG1) is an epigenetically regulated lncRNA that represents a promising therapeutic target due to its tumor suppressor properties regulating the p53-mediated DNA damage and the intracellular localization of the oncogenic YBX1 protein. However, to translate this finding into the clinic as a gene therapy, it is important to develop effective carriers able to deliver exogenous lncRNAs to the targeted cancer cells. Here, we propose the use of biocompatible sphingomyelin nanosystems comprising DOTAP (DSNs) to carry and deliver a plasmid vector encoding for TP53TG1 (pc(TP53TG1)-DSNs) to a colorectal cancer cell line (HCT-116). DSNs presented a high association capacity and convenient physicochemical properties. In addition, pc(TP53TG1)-DSNs showed anti-tumor activities in vitro, specifically a decrease in the proliferation rate, a diminished colony-forming capacity, and hampered migration and invasiveness of the treated cancer cells. Consequently, the proposed strategy displays a high potential as a therapeutic approach for colorectal cancer.

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Curcumin Regulates ERCC1 Expression and Enhances Oxaliplatin Sensitivity in Resistant Colorectal Cancer Cells through Its Effects on miR-409-3p

Evidence-based Complementary and Alternative Medicine ◽

10.1155/2020/8394574 ◽

2020 ◽

Vol 2020 ◽

pp. 1-16

Author(s):

Wei Han ◽

Hongli Yin ◽

Hao Ma ◽

Yi Wang ◽

Desong Kong ◽

...

Keyword(s):

Colorectal Cancer ◽

Drug Resistance ◽

Cancer Cells ◽

Resistance Mechanism ◽

Cancer Cell Line ◽

Colorectal Cancer Cell Line ◽

Pcr Analysis ◽

Colorectal Cancer Cells ◽

Ercc1 Expression

Background. Oxaliplatin (L-OHP) resistance is a major obstacle to the effective treatment of colorectal cancer. The resistance mechanism(s) of colorectal tumors to L-OHP may be related to the regulation of ERCC1 by cancer-expressed miRNAs, but no in-depth studies on the miRNAs that affect drug resistance have been performed. Curcumin (Cur) can reverse the drug resistance of cancer cells, but its effects on ERCC1 expression and miRNA profiles in colorectal cancer have not been studied. Methods. To study the regulation effect of curcumin on ERCC1 expression and its effects on miRNAs, the L-OHP-resistant colorectal cancer cell line HCT116/L-OHP was established. MTT assays were used to evaluate cell proliferation. Flow cytometry was used to investigate apoptotic induction. Western blot and RT-PCR analysis were used to evaluate the expression of drug-associated ERCC1, Bcl-2, GST-π, MRP, P-gp, and survivin. Results. HCT116//L-OHP cell lines were successfully established. The combination of L-OHP and curcumin could reduce L-OHP resistance in vitro. In addition, combination therapy inhibited the expression of ERCC1, Bcl-2, GST-π, MRP, P-gp, and survivin at the mRNA and protein level. Curcumin was found to inhibit ERCC1 through its ability to modulate miR-409-3p. Conclusion. Curcumin can overcome L-OHP resistance in colorectal cancer cells through its effects on miR-409-3p mediated ERCC1 expression.

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The POLD1R689W variant increases the sensitivity of colorectal cancer cells to ATR and CHK1 inhibitors

Scientific Reports ◽

10.1038/s41598-020-76033-1 ◽

2020 ◽

Vol 10 (1) ◽

Author(s):

Albert Job ◽

Marina Tatura ◽

Cora Schäfer ◽

Veronika Lutz ◽

Hanna Schneider ◽

...

Keyword(s):

Colorectal Cancer ◽

Dna Replication ◽

Cancer Cells ◽

Cancer Cell Line ◽

Colorectal Cancer Cell Line ◽

Variants Of Unknown Significance ◽

Pathway Activation ◽

Synthetic Lethal Interactions

Abstract Inhibition of the kinase ATR, a central regulator of the DNA damage response, eliminates subsets of cancer cells in certain tumors. As previously shown, this is at least partly attributable to synthetic lethal interactions between ATR and POLD1, the catalytic subunit of the polymerase δ. Various POLD1 variants have been found in colorectal cancer, but their significance as therapeutic targets for ATR pathway inhibition remains unknown. Using CRISPR/Cas9 in the colorectal cancer cell line DLD-1, which harbors four POLD1 variants, we established heterozygous POLD1-knockout clones with exclusive expression of distinct variants to determine the functional relevance of these variants individually by assessing their impact on ATR pathway activation, DNA replication, and cellular sensitivity to inhibition of ATR or its effector kinase CHK1. Of the four variants analyzed, only POLD1R689W affected POLD1 function, as demonstrated by compensatory ATR pathway activation and impaired DNA replication. Upon treatment with ATR or CHK1 inhibitors, POLD1R689W strongly decreased cell survival in vitro, which was attributable at least partly to S phase impairment and apoptosis. Similarly, treatment with the ATR inhibitor AZD6738 inhibited growth of murine xenograft tumors, harboring the POLD1R689W variant, in vivo. Our POLD1-knockout model thus complements algorithm-based models to predict the pathogenicity of tumor-specific variants of unknown significance and illustrates a novel and potentially clinically relevant therapeutic approach using ATR/CHK1 inhibitors in POLD1-deficient tumors.

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Effect of EGFR inhibition on HER3/PI3K activation by feedback induction of ErbB heterodimers in cetuximab-sensitive colon cancer cells.

Journal of Clinical Oncology ◽

10.1200/jco.2013.31.15_suppl.3626 ◽

2013 ◽

Vol 31 (15_suppl) ◽

pp. 3626-3626

Author(s):

Bart Jacobs ◽

Loredana Vecchione ◽

Nicholas Hoe ◽

Jef De Schutter ◽

Bart Biesmans ◽

...

Keyword(s):

Colorectal Cancer ◽

Cancer Cells ◽

Cell Line ◽

Cancer Cell Line ◽

Therapeutic Index ◽

Network Activity ◽

Colorectal Cancer Cell Line ◽

Egfr Inhibition ◽

Downstream Signaling ◽

Protein Levels

3626 Background: Although cetuximab treatment has been successful for the treatment of KRAS wild-type colorectal cancers, complete remissions are rarely seen in patients, leading ultimately to resistance. We hypothesized that in cetuximab-sensitive patients, the ErbB network is insufficiently targeted since network plasticity may occur. Methods: We have used EGFR-sensitive colorectal cancer cells and investigated ErbB network activity and adaptations by RTK arrays, western blot, and Collaborative Enzyme Enhance Reactive immunoassay (CEER). These were complemented by ErbB heterodimerization (HER2:3, HER1:2, HER1:3, HER3:PI3K) assays using CEER. Effects on cell survival were measured using colony formation assay. In addition to the EGFR sensitive cell line, 200 clinical colorectal cancer (CRC) samples were profiled utilizing CEER for RTKs, downstream signaling, and heterodimerization. Results: EGFR and downstream signaling proteins AKT, ERK, and RSK were potently inhibited by cetuximab or gefitinib at 24h of treatment in EGFR sensitive colorectal cancer cell line. At 24h of treatment, we observed approximately 2 folds increase in total HER2 and HER3 protein levels, 2.7 folds in phosphorylated HER3, and 5.4 folds in HER2:HER3 heterodimer formation. Concurrently, increased in ErbB heterodimer formation was accompanied by 5 folds increase in PI3K binding to HER3, resulting in enhanced HER3 signaling, with increase in AKT, ERK, and RSK. Co-treatment of these cells with cetuximab and HER2 inhibitor Trastuzumab or by treatment with lapatinib blocked the induction of HER2:HER3 heterodimer, HER3 phosphorylation, and PI3K binding to HER3. In 30% of the 200 clinical colorectal cancer samples profiled, we observed an increased in phosphorylated HER3, formation of HER2:HER3 and HER3:PI3K heterodimers along with HER1 activation (KRAS WT). Conclusions: Combination of HER2 inhibitor with an EGFR inhibitor could potentially increase the therapeutic index in cetuximab sensitive patients, and suppress activation of feedback mechanisms upon EGFR inhibition. Current findings suggest that CRC patients with similar profile would benefit from these combination therapies.

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Synergistic Effect of Free and Nano-encapsulated Chrysin-Curcumin on Inhibition of hTERT Gene Expression in SW480 Colorectal Cancer Cell Line

Drug Research ◽

10.1055/s-0043-121338 ◽

2018 ◽

Vol 68 (06) ◽

pp. 335-343 ◽

Cited By ~ 7

Author(s):

Raana Bagheri ◽

Zohreh Sanaat ◽

Nosratollah Zarghami

Keyword(s):

Gene Expression ◽

Colorectal Cancer ◽

Synergistic Effect ◽

Cancer Cells ◽

Cell Line ◽

Cancer Cell ◽

Cancer Cell Line ◽

Colorectal Cancer Cell Line ◽

Htert Gene ◽

Sw480 Cells

Abstract Background Telomerase is known as a global therapeutic target in cancer cells due to its main role in tumorigenesis. Nowadays, it is proposed new treatment methods based on molecular target therapy by bioactive substances such as curcumin and chrysin with fewer side effects than other chemical drugs. But due to their low aqueous solubility and high clearance in the bloodstream it can be used of nanoparticles to increase their half-life and biocompatibility of them. Therefore, the goal of this study was to evaluate the effect of Chrysin-Curcumin on the expression of telomerase gene in SW480 colorectal cancer cell line. Material and method PLGA-PEG nanoparticles synthesized and were confirmed using by the scanning electron microscope (SEM) and FTIR Spectroscopy. After treatment of SW480 cells by curcumin and chrysin loaded nanoparticles, their toxicity to cancer cells, was evaluated by MTT. Then, the inhibition of hTERT gene expression was measured using qRT-PCR method. Result The results of MTT test showed nanocapsulated curcumin and chrysin compared with free forms of these compounds have high synergistic effect on sw480 cells. Also, real time-PCR showed significant decrease in hTERT gene expression in SW480 cells that treated with nano-curcumin and nano-chrysin compare to untreated cells. Conclusion Nano-encapsulation of curcumin and chrysin enhanced delivery of these compounds to SW480 colorectal cancer cells and therefore it can be conclude that PLGA-PEG nanoparticles promote anticancer effects of curcumin-chrysin by increasing bioavailability and the solubility of these drugs.

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Antiproliferative activity and caspase enhancement properties of Annona muricata leaves extract against colorectal cancer cells

Medical Journal of Indonesia ◽

10.13181/mji.v25i3.1449 ◽

2016 ◽

Vol 25 (3) ◽

pp. 136-42

Author(s):

Lili Indrawati ◽

Purwantyastuti Ascobat ◽

Budiman Bela ◽

Murdani Abdullah ◽

Ingrid S. Surono ◽

...

Keyword(s):

Colorectal Cancer ◽

Cell Line ◽

Cancer Cell ◽

Water Extract ◽

Cancer Cell Line ◽

Colorectal Cancer Cell ◽

Colorectal Cancer Cell Line ◽

Annona Muricata ◽

Caspase 9

Background: The prevalence of colorectal cancer is rising in Asia including Indonesia. Annona muricata tea leaves, that is traditionally used for maintaining health, and lately being used by cancer patients. The objectives of this study is to investigate its effects in human colorectal cancer cell in vitro and ex vivo.Methods: Thirty patients with colorectal cancer (CRC) were enrolled in a randomized double-blind placebo-controlled trial. They were equally divided into two groups: those treated with 300 mg A. muricata leaf extract and placebo daily for 8 weeks. Serum from supplemented CRC patients of both groups was compared for caspase 9 and caspase 8 enhancement activity. Antiproliferative effect of water extract of A. muricata leaves and its fractions were evaluated against colorectal cancer cell line (DLD-1 and COLO 205) compared with 5-fluorouracil and placebo, the dose range was 62.5-2,000 µg/mL. Method used was 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Data were analyzed by Mann-Whitney U test. The p value was set at 0.05.Results: Ethanol-soluble fraction of A. muricata leaves extract water extract (ESFAM) leaves extract had cytotoxicity effects on DLD-1 as well as COLO 205 cell line, as shown by the lower IC50 compared to 5-fluorouracil and placebo, 20.59 μg/mL and 654.9μg/mL, respectively. Serum of subjects supplemented with extract significantly induced caspase 9 (p=0.001) activity of DLD-1 colorectal cancer cell line, but not for caspase 8 activity (p=0.372).Conclusion: The study's results suggest the cytotoxicity potential of A. muricata leaves extract in in vitro and ex vivo studies.

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Cytotoxic, genotoxic and apoptotic effects of Viburnum opulus on colon cancer cells: an in vitro study

Turkish Journal of Biochemistry ◽

10.1515/tjb-2020-0182 ◽

2020 ◽

Vol 0 (0) ◽

Author(s):

Kubra Bozali ◽

Eray Metin Guler ◽

Ahmet Sadik Gulgec ◽

Abdurrahim Kocyigit

Keyword(s):

Colorectal Cancer ◽

Dna Damage ◽

Cell Viability ◽

Cancer Cells ◽

Cancer Cell ◽

Methanolic Extract ◽

Cancer Cell Line ◽

Dependent Manner ◽

Apoptotic Effects

AbstractObjectiveIntake of various fruits is quite significant for maintaining the human body, due to their supply of useful constituents. V. opulus has been found to have outstanding antioxidant activity while showing a pro-oxidant effect at high doses. Due to this feature, V. opulus would be anticipated to have a healing impact on cancer treatment. In this study, it has been proposed to examine the cytotoxic, genotoxic, and apoptotic effects of V. opulus on human colorectal cancer cell.MethodDifferent concentrations of V. opulus methanolic extract (5–2000 μg/mL) were incubated for 24 h with colorectal cancer cell line (Lovo). The cell viability, intracellular reactive oxygen species (iROS), DNA damage, and apoptosis were measured after incubation.ResultsThe obtained results of this research demonstrate decreased cell viability and increased DNA damage, iROS, and apoptosis levels of V. opulus in Lovo cells in a concentration-dependent manner in the range of 14.88–52.06%. There were strong positive relationships between apoptosis, genotoxicity, and cytotoxicity in V. opulus methanolic extract treated cancer cell line.DiscussionThis in vitro research clearly demonstrated that V. opulus methanolic extract induces DNA damage, apoptosis, and cytotoxicity in a dose-dependent manner in cancer cells due to its pro-oxidant activity.ConclusionAlthough in vitro results are favorable, in vivo and further studies are needed.

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Regulation Network of Colorectal-Cancer-Specific Enhancers in the Progression of Colorectal Cancer

International Journal of Molecular Sciences ◽

10.3390/ijms22158337 ◽

2021 ◽

Vol 22 (15) ◽

pp. 8337

Author(s):

Bohan Chen ◽

Yiping Ma ◽

Jinfang Bi ◽

Wenbin Wang ◽

Anshun He ◽

...

Keyword(s):

Colorectal Cancer ◽

Cancer Cells ◽

Cancer Progression ◽

Target Genes ◽

Colorectal Cancer Cell Line ◽

Tissue Samples ◽

Colon Cells ◽

Colorectal Cancer Cells ◽

Regulation Network

Enhancers regulate multiple genes via higher-order chromatin structures, and they further affect cancer progression. Epigenetic changes in cancer cells activate several cancer-specific enhancers that are silenced in normal cells. These cancer-specific enhancers are potential therapeutic targets of cancer. However, the functions and regulation networks of colorectal-cancer-specific enhancers are still unknown. In this study, we profile colorectal-cancer-specific enhancers and reveal their regulation network through the analysis of HiChIP data that were derived from a colorectal cancer cell line and Hi-C and RNA-seq data that were derived from tissue samples by in silico analysis and in vitro experiments. Enhancer–promoter loops in colorectal cancer cells containing colorectal-cancer-specific enhancers are involved in more than 50% of the topological associated domains (TADs) changed in colorectal cancer cells compared to normal colon cells. In addition, colorectal-cancer-specific enhancers interact with 152 genes that are significantly and highly expressed in colorectal cancer cells. These colorectal-cancer-specific enhancer target genes include ITGB4, RECQL4, MSLN, and GDF15. We propose that the regulation network of colorectal-cancer-specific enhancers plays an important role in the progression of colorectal cancer.

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Proteomic characterization of GSK3β knockout shows altered cell adhesion and metabolic pathway utilisation in colorectal cancer cells.

10.1101/2021.01.26.428222 ◽

2021 ◽

Author(s):

E.H. Bowler-Barnett ◽

F. D. Martinez-Garcia ◽

M. Sherwood ◽

S. Weston ◽

Y. Wang ◽

...

Keyword(s):

Colorectal Cancer ◽

Cell Adhesion ◽

Cancer Cells ◽

Cancer Cell ◽

Cell Model ◽

Cancer Cell Line ◽

Glycogen Metabolism ◽

Colorectal Cancer Cell ◽

Colorectal Cancer Cell Line ◽

Colorectal Cancer Cells

ABSTRACTGlycogen-specific kinase (GSK3β) is an integral regulator of the Wnt signalling pathway as well as many other diverse signalling pathways and processes. Dys-regulation of GSK3β is implicated in many different pathologies, including neurodegenerative disorders as well as many different tumour types. In the context of tumour development, GSK3β has been shown to play both oncogenic and tumour suppressor roles, depending upon tissue, signalling environment or disease progression. Although multiple substrates of the GSK3β kinase have been identified, the wider protein networks within which GSK3β participates are not well known, and the consequences of these interactions not well understood. In this study, LC-MS/MS expression analysis was performed using knockout GSK3β colorectal cancer cells and isogenic controls in colorectal cancer cell lines carrying dominant stabilizing mutations of β-Catenin. Consistent with the role GSK3β, we found that β-Catenin levels and canonical Wnt activity are unaffected by knockout of GSK3β and therefore use this knockout cell model to identify other processes in which GSK3β is implicated. Quantitative proteomic analysis revealed perturbation of proteins involved in cell-cell adhesion, and we characterize the phenotype and altered proteomic profiles associated with this. We also characterize the perturbation of metabolic pathways resulting from GSK3β knockout and identify defects in glycogen metabolism. In summary, using a precision colorectal cancer cell-line knockout model with constitutively activated β-Catenin we are able to identify several of the diverse pathways and processes associated with GSK3β function.

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A Comparison of the Anti-Cancer Effects of Free and PLGA-PAA Encapsulated Hydroxytyrosol on the HT-29 Colorectal Cancer Cell Line

Anti-Cancer Agents in Medicinal Chemistry ◽

10.2174/1871520621666210308095712 ◽

2021 ◽

Vol 21 ◽

Author(s):

Nasrin S. Sani ◽

Habib Onsori ◽

Somayeh Akrami ◽

Mohammad Rahmati

Keyword(s):

Colorectal Cancer ◽

Cancer Cells ◽

Cell Line ◽

Cancer Cell ◽

Cancer Cell Line ◽

Colorectal Cancer Cell ◽

Colorectal Cancer Cell Line ◽

Cycle Arrest ◽

Anti Cancer ◽

Colorectal Cancer Cells

Background: Hydroxytyrosol is one of the phenolic compounds of olive oil and can induce anti-cancer effects on the colorectal cancer cells. Objective: The aim of the present study was to evaluate the free hydroxytyrosol and nano-capsulated hydroxytyrosol effects on the cell cycle arrest in HT-29 colorectal cancer cell line. Methods: The nano-capsulated hydroxytyrosol was synthesized in poly lactide-co-glycolide-co-polyacrylic acid (PLGA-PAA) copolymer. MTT assay was performed to evaluate the anti- proliferative and anti-tumor effects of the free hydroxytyrosol and nano-capsulated hydroxytyrosol. Finally, the relative expression of CDKN1A, CDKN1B and CCND1 genes was evaluated in the control and treated colorectal cancer cells by using Real-Time PCR. Results: The obtained results from the MTT assay showed that the cytotoxic effects of the nano-capsulated hydroxytyrosol on the colorectal cancer cell line (IC50= 6PPM) was significantly more than free hydroxytyrosol (IC50= 12PPM) after 72h. Also, nano-capsulated hydroxytyrosol showed more significant effects on the up-regulation of CDKN1A and CDKN1B genes, and down-regulation of the CCND1 gene in the colorectal cancer cells. Conclusion: In conclusion, the present study showed that the hydroxytyrosol led to die the colorectal cancer cell through the cell cycle arrest. Also, the PLGA-PAA copolymer dramatically caused to increase the cytotoxic effects of the hydroxytyrosol on the colorectal cancer cells.

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Inhibition of Colorectal Cancer Cell Line CaCo-2 by Essential Oil of Eucalyptus camaldulensis Through Induction of Apoptosis

ACTA MEDICA IRANICA ◽

10.18502/acta.v58i6.4051 ◽

2020 ◽

Author(s):

Elham Taheri ◽

Saba Ghorbani ◽

Maryam Safi ◽

Nasrin Seyyed Sani ◽

Fatemeh Firouzi Amoodizaj ◽

...

Keyword(s):

Colorectal Cancer ◽

Antioxidant Activity ◽

Essential Oil ◽

Cancer Cells ◽

Cell Line ◽

Cancer Cell Line ◽

Eucalyptus Camaldulensis ◽

Colorectal Cancer Cell Line ◽

Dependent Manner ◽

Induction Of Apoptosis

Treatment of colorectal cancer is one of the important challenges due to the increase of resistance to chemotherapeutic drugs. Isolated natural compounds from medicinal plants and other sources often are used as novel drugs for treatment of various human cancer. The aim of this study was to investigate the antioxidant and anticancer activity of Eucalyptus camaldulensis essential oil on colorectal cancer cell line Caco-2. The antioxidant activity of extracted E. camaldulensis essential oil (1000, 800, 400, 200, 100, 50, 25, 12.5, 6, and 3 μg/mL) was evaluated by free radicals inactivation method. Moreover, MTT assay was used to examine the cytotoxic effects of E. camaldulensis essential oil on the Caco-2 cell line. The mRNA expression of BAX and BCL-2 genes was studied using quantitative Real-Time PCR method, in treated cancer cells compared to untreated cells. We indicated a significant, impressive antioxidant activity in 1000 μg/mL of E. camaldulensis essential oil, in a concentration-dependent manner. In addition, we found that this product exerted a cytotoxic effect on cancer cells when 100 μg/mL concentration was considered as half-maximal inhibitory concentration (IC50). Also, the expression of BAX and BCL-2 genes were significantly upregulated and downregulated, respectively, in the treated Caco-2 cells with E. camaldulensis essential oil. In conclusion, our study showed significant antioxidant and anticancer activity in E. camaldulensis essential oil in a concentration and time-dependent manner, which may be due to the reduction of free radicals and induction of apoptosis process in colorectal cancer cells.

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