scholarly journals P09.11 TLR3 suppresses colorectal carcinogenesis, presumably through up-regulation of T-cell attracting CXC chemokines

2020 ◽  
Vol 8 (Suppl 2) ◽  
pp. A57.2-A58
Author(s):  
A Sichler ◽  
M Frey ◽  
W Johannes ◽  
K Janssen
Keyword(s):  
Colorectal Cancer ◽  
Colon Cancer ◽  
T Cell ◽  
Cell Lines ◽  
Colon Cancer Cell ◽  
Human Colorectal Cancer ◽  
T Cell Infiltration ◽  
Tlr3 Expression ◽  
Colon Cancer Cell Lines

BackgroundToll-like-receptors (TLRs) are main components of the innate immune system which recognize endogenous or pathogen-associated molecular ‘danger patterns’. Previous findings from us and others highlighted a role of TLRs in the formation of tumors. However, TLRs may have differing roles in immune and cancer cells, and the underlying mechanisms are still unclear. TLR downstream signaling is mediated by two adapter proteins; MyD88 (myeloid differentiation primary response gene 88) and TRIF (TIR-domain-containing adapter-inducing interferon-β). The MyD88-mediated signaling pathway is activated by all TLRs, except TLR3. We could show previously that it leads to the initiation of oncogenic, proliferative and pro-inflammatory responses in colorectal cancer. The endosomal receptor TLR3, in contrast, solely depends on TRIF. It recognizes viral, microbial and endogenous dsRNA leading to production of type-I interferon and chemokines, and induces apoptosis. The role of TRIF dependent TLR3 signaling in colorectal cancer is still disputed. Within this study, we show tumor-suppressive TLR3 functions prevailing over oncogenic effects in colorectal cancer.Materials and MethodsTLR3-deficient colon cancer cell lines were engineered by CRISPR-Cas9. Genetically modified mouse models were generated, based on a ‘switch-on mutagenesis’ approach, with global inactivation of Tlr3 or TRIF (Ticam1), allowing tissue specific re-expression based on Cre recombination. The mice were interbred with the Apc1638N mouse model for digestive cancer. Furthermore, clinical significance of TLR3 expression levels was assessed in human colorectal cancer tissue samples from our clinic (n=81) and from TCGA datasets. A putative correlation between intratumoral TLR3 expression and T-cell infiltration is currently analyzed.ResultsGlobal deficiency of murine Tlr3, or TRIF, induced significantly increased digestive tumor formation, associated with increased morbidity indicating a tumor suppressive role. Coherently, TLR3 expression is highly significantly decreased in human colorectal cancer compared to normal mucosa, significantly correlated with poor survival. TLR3 deficient cell lines show reduced migration and slightly declined proliferation suggesting an oncogenic role on the cell-autonomous level. Nevertheless, gene expression analysis revealed that the dsRNA induced expression of T-cell attracting cytokines CXCL10 and CXCL11 in colon cancer cell lines is exclusively dependent on TLR3. These chemokines were shown to favor a TH1-type antitumoral response.ConclusionsTLR3 favors tumor suppression in vivo, presumably resulting from non-cell-autonomous factors such as the production of CXCL10 and 11 and resulting T-cell infiltration. This may outweigh the putative cell-autonomous oncogenic functions of TLR3 deficiency.Disclosure InformationA. Sichler: None. M. Frey: None. W. Johannes: None. K. Janssen: None.

scholarly journals Anticancer effects of bifidobacteria on colon cancer cell lines

2021 ◽  
Vol 21 (1) ◽  
Author(s):  
Zeinab Faghfoori ◽  
Mohammad Hasan Faghfoori ◽  
Amir Saber ◽  
Azimeh Izadi ◽  
Ahmad Yari Khosroushahi
Keyword(s):  
Colorectal Cancer ◽  
Colon Cancer ◽  
Cancer Cells ◽  
Cell Lines ◽  
Cancer Cell ◽  
Colon Cancer Cell ◽  
Rt Pcr ◽  
Anticancer Effects ◽  
Colon Cancer Cell Lines ◽  
Apoptotic Genes

Abstract Background Colorectal cancer (CRC), with a growing incidence trend worldwide, is resistant to apoptosis and has uncontrolled proliferation. It is recently reported that probiotic microorganisms exert anticancer effects. The genus Bifidobacterium, one of the dominant bacterial populations in the gastrointestinal tract, has received increasing attention because of widespread interest in using it as health-promoting microorganisms. Therefore, the present study aimed to assess the apoptotic effects of some bifidobacteria species on colon cancer cell lines. Methods The cytotoxicity evaluations performed using MTT assay and FACS-flow cytometry tests. Also, the effects of five species of bifidobacteria secretion metabolites on the expression level of anti- or pro-apoptotic genes including BAD, Bcl-2, Caspase-3, Caspase-8, Caspase-9, and Fas-R studied by real-time polymerase chain reaction (RT-PCR) method. Results The cell-free supernatant of all studied bifidobacteria significantly decreased the survival rates of colon cancer cells compared with control groups. Flow cytometric and RT-PCR results indicated that apoptosis is induced by bifidobacteria secretion metabolites and the mechanism for the action of bifidobacteria species in CRC prevention could be down-regulation and up-regulation of anti-apoptotic and, pro-apoptotic genes. Conclusions In the present study, different bifidobacteria species showed anticancer activity on colorectal cancer cells through down-regulation and up-regulation of anti-apoptotic and pro-apoptotic genes. However, further studies are required to clarify the exact mechanism of apoptosis induction by bifidobacteria species.

Prediction and prognostic validation of colorectal cancer therapeutic effect of WM-S1-030 using pre-clinical CTC animal model.

2020 ◽  
Vol 38 (15_suppl) ◽  
pp. e15557-e15557
Author(s):  
Hae Ung Lee ◽  
Jai-Hee Moon ◽  
Joseph Kim ◽  
Seong Keun Kim ◽  
Jae Sik Shin ◽  
...  
Keyword(s):  
Colorectal Cancer ◽  
Colon Cancer ◽  
Animal Model ◽  
Cancer Patients ◽  
Cell Lines ◽  
Tumor Size ◽  
Peripheral Blood ◽  
Colon Cancer Cell ◽  
Nsg Mice ◽  
Colon Cancer Cell Lines

e15557 Background: CTC (Circulating tumor cells) has the advantage of being able to effectively investigate non-invasive diagnosis, prediction, and prognosis for non-surgical carcinoma or tissue sample collection. We have developed WM-S1-030 (WMBIO) as a novel inhibitor for mtRTK (mutant receptor tyrosine kinase) identified important targets for cancer progression and metastasis in various cancers, including colorectal cancer, and have tried to prove the possibility of CTC diagnosis, prediction and prognosis of WM-S1-030 using tumor animal models. Methods: Six of colon cancer cell lines expressing mtRTK (pTyr-mtRTK) were transplanted into Balb.c nude mice and NSG mice to collect peripheral blood when the tumor size reached 1000 mm3. CTCs were quantified in peripheral blood using the Smart Biopsy cell isolator (Cytogen). The expression of pTyr-mtRTK in CTCs was analyzed via the Smart Biopsy Image Analyzer (Cytogen). Colon cancer cell lines expressing pTyr-mtRTK were transplanted into NSG mice and we examined the tumor suppression, the enumeration of CTCs and pTyr-mtRTK expression in CTCs after administration of WM-S1-030. We also identified CTCs in peripheral blood of colon cancer patients and analyzed expression of pTyr-mtRTK in CTCs through Smart Biopsy system. Results: As results of the enumeration of CTCs after tumor transplantation into Balb.c nude mice and NSG mice, the number of CTC was 8 ~ 20 times higher peripheral blood in the NSG model, which indicates innate immune responses are critical for the survival of CTCs. Half of CTCs isolated from the blood expressed pTyr-mtRTK. The tumor size, the number of CTCs, and the expression of pTyr-mtRTK in CTCs were examined after the administration of WM-S1-030. The volume of tumor decreased by 67% in the WM-S1-030 group compared to the vehicle. The number of CTCs was reduced by approximately three times after the treatment of WM-S1-030 and also the number of CTCs expressing pTyr-mtRTK was significantly decreased. CTC detection and pTyr-mtRTK expression analysis in the peripheral blood of colon cancer patients detected 9 CTCs per 10 ml of peripheral blood, of which 11 % of CTCs representing pTyr-mtRTK were confirmed. Conclusions: WM-S1-030 was developed and confirmed in the CTC-based animal model, which implies that the analysis of CTCs is the powerful tool for predicting/ prognosing WM-S1-030 in the pre-clinical analysis. Currently, we are conducting clinical diagnostic validation using CTC in the blood of colorectal cancer patients and preparing for clinical trial of WM-S1-030.

Dual Targeting of the Epidermal Growth Factor Receptor Using the Combination of Cetuximab and Erlotinib: Preclinical Evaluation and Results of the Phase II DUX Study in Chemotherapy-Refractory, Advanced Colorectal Cancer

2012 ◽  
Vol 30 (13) ◽  
pp. 1505-1512 ◽  
Author(s):  
Andrew J. Weickhardt ◽  
Tim J. Price ◽  
Geoff Chong ◽  
Val Gebski ◽  
Nick Pavlakis ◽  
...  
Keyword(s):  
Colorectal Cancer ◽  
Colon Cancer ◽  
Epidermal Growth Factor Receptor ◽  
Cell Lines ◽  
Cancer Cell ◽  
Growth Factor Receptor ◽  
Cancer Cell Lines ◽  
Colon Cancer Cell ◽  
Colon Cancer Cell Lines ◽  
Epidermal Growth

Purpose This preclinical and phase II study evaluated the efficacy and safety of the combination of cetuximab and erlotinib in metastatic colorectal cancer (mCRC). Patients and Methods The activity and mechanism of action of the combination of cetuximab plus erlotinib were investigated in vitro in colorectal cancer cell lines. In the clinical study, patients with chemotherapy-refractory mCRC were treated with cetuximab 400 mg/m2 as a loading dose and then weekly cetuximab 250 mg/m2 with erlotinib 100 mg orally daily. The primary end point was response rate (RR), which was evaluated separately in KRAS wild-type (WT) versus KRAS mutant tumors. Secondary end points included toxicity, progression-free survival (PFS), and overall survival. Target accrual was 50 patients, with a one-stage design. Results Preclinical studies demonstrated synergistic activity of cetuximab and erlotinib cotreatment on growth inhibition of colon cancer cell lines both as a result of enhanced inhibition of the epidermal growth factor receptor pathway and differential effects on STAT3. In the clinical study, 50 patients were enrolled, with 48 patients evaluable for response. The overall RR was 31% (95% CI, 26% to 57%), with a median PFS of 4.6 months (95% CI, 2.8 to 5.6 months). RR was 41% (95% CI, 26% to 57%) in KRAS WT tumors, with a median PFS of 5.6 months (95% CI, 2.9 to 5.6 months). There was no response in 11 patients with KRAS mutations. Frequent grade 3 and 4 toxicities were rash (48%), hypomagnesaemia (18%), and fatigue (10%). Conclusion The combination of cetuximab and erlotinib synergistically inhibits growth of colon cancer cell lines, achieves promising efficacy in patients with KRAS WT mCRC, and merits evaluation in further randomized studies.

scholarly journals MicroRNA silencing of CNN1 and CNN2 protein family members regulates biology processes in colorectal cancer by targeting the p53 signal pathway

2020 ◽  
Author(s):  
Fuda Huang ◽  
Mingwei Wei ◽  
Anmin Wang ◽  
Ya Zhang ◽  
Zebang Qin ◽  
...  
Keyword(s):  
Colorectal Cancer ◽  
Cell Cycle ◽  
Colon Cancer ◽  
Cell Lines ◽  
Cancer Cell ◽  
Cancer Cell Lines ◽  
Colon Cancer Cell ◽  
Expression Levels ◽  
Colon Cancer Cell Lines ◽  
Cancer Tissues

Abstract BackgroundCalponin was first defined as a striated muscle troponin T-like protein that binds actin thin filaments to regulate smooth muscle contraction. There are few studies of CNN1 and CNN2 in colorectal cancer, and the roles these two genes play in colorectal cancer cell lines and the mechanisms by which they act are unknown.MethodsWe used immunohistochemistry to identify expression of the two genes in the cancer tissues. RT-PCR was used to measure expression levels of microRNA. W performed western blots to measure changes in signaling pathways in the context of expression interference.Meanwhile, the same method was used to measure binding relationship between the two genes and key pathway proteins. To determine the relationship between microRNA and gene mRNA, we used the reporter gene method. We used the chi-square and t-test methods to analyze the significance and correlations of the data.Results and conclusionsExpression levels of CNN1 were lower in colon cancer tissues than in normal mucosal tissues. After downregulating CNN1, the cell cycle in colon cancer cell lines progressed quickly, and the expression of related pathway proteins also increased. Expression levels of CNN2 were higher in colon cancer tissues, and its downregulation significantly inhibited cell cycle progression in colon cancer cell lines. We confirmed correlations between the expression of microRNA and CNN2 using data analysis.Bars indicate ± standard errors.*p < 0.05; **p < 0.01 compared with the control. The inhibition of the expression of CNN2 mRNA using microRNA was confirmed using western blot. The combination of the two at the mechanism level was also demonstrated using the reporter gene method.

scholarly journals Anticancer effects of Bifidobacteria on colon cancer cell lines

2021 ◽  
Author(s):  
Zeinab Faghfoori ◽  
Mohammad Hasan Faghfoori ◽  
Amir Saber ◽  
Azimeh Izadi ◽  
Ahmad Yari Khosroushahi
Keyword(s):  
Colorectal Cancer ◽  
Colon Cancer ◽  
Cancer Cells ◽  
Cell Lines ◽  
Cancer Cell ◽  
Colon Cancer Cell ◽  
Rt Pcr ◽  
Anticancer Effects ◽  
Colon Cancer Cell Lines ◽  
Apoptotic Genes

Abstract Background: Colorectal cancer (CRC), with a growing incidence trend worldwide, is resistant to apoptosis and have the uncontrolled proliferation. It is recently reported that probiotic microorganisms exert anticancer effects. The genus Bifidobacterium, one of the dominant bacterial populations in the gastrointestinal tract, has received increasing attention because of widespread interest in using as health-promoting microorganisms. Therefore, the present study aimed to assess the apoptotic effects of some bifidobacteria species on colon cancer cell lines.Methods: The cytotoxicity evaluations performed using MTT assay and FACS-flow cytometry tests. Also, the effects of five species of bifidobacteria secretion metabolites on the expression level of anti- or pro-apoptotic genes including BAD, Bcl-2, Caspase-3, Caspase-8, Caspase-9, and Fas-R studied by real-time polymerase chain reaction (RT-PCR) method. Results: The cell-free supernatant of all studied bifidobacteria significantly decreased the survival rates of colon cancer cells compared with control groups. Flow cytometric and RT-PCR results indicated that apoptosis is induced by bifidobacteria secretion metabolites and the mechanism for the action of bifidobacteria species in CRC prevention could be down-regulation and up-regulation of anti-apoptotic and, pro-apoptotic genes.Conclusions: In the present study, different bifidobacteria species showed anticancer activity on colorectal cancer cells through down-regulation and up-regulation of anti-apoptotic and pro-apoptotic genes. However, further studies are required to clarify the exact mechanism of apoptosis induction by bifidobacteria species.

scholarly journals Mechanism of apoptosis induced by S100A8/A9 in colon cancer cell lines: the role of ROS and the effect of metal ions

2004 ◽  
Vol 76 (1) ◽  
pp. 169-175 ◽  
Author(s):  
Saeid Ghavami ◽  
Claus Kerkhoff ◽  
Marek Los ◽  
Mohammad Hashemi ◽  
Clemens Sorg ◽  
...  
Keyword(s):  
Colon Cancer ◽  
Metal Ions ◽  
Cell Lines ◽  
Cancer Cell ◽  
Cancer Cell Lines ◽  
Colon Cancer Cell ◽  
Colon Cancer Cell Lines
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