Seyed Mohammad Javadzadeh; Mohsen Keykhosravi; Mohsen Tehrani; Hossein Asgarian-Omran; Mohsen Rashidi; Hadi Hossein-Nattaj; Laleh Vahedi-Larijani; Abolghasem Ajami
Abstract
Background: Innate Lymphoid Cells (ILCs) promote tissue homeostasis, contribute to the immune defense mechanisms, and play important roles in the initiation of immune responses and chronic inflammation. Objective: To understand the roles of innate lymphoid cells in the pathophysiology of colorectal ...
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Background: Innate Lymphoid Cells (ILCs) promote tissue homeostasis, contribute to the immune defense mechanisms, and play important roles in the initiation of immune responses and chronic inflammation. Objective: To understand the roles of innate lymphoid cells in the pathophysiology of colorectal cancer (CRC) in the mouse model. Methods: CRC was induced using azoxymethane (AOM) and dextran sulfate sodium (DSS) in Balb/c mice (the chemically induced group=18 mice), or orthotopic injection of CT-26 cell line into the colon of another set of Balb/c mice (the orthotopic group=14mice). Normal saline was injected into 18 mice, as the sham group. After 80 days, the chemically induced group was divided into two subgroups, dysplasia (8 mice) and reparative change (10 mice), based on pathological examinations. The frequencies of ILC1, 2, and 3 were then measured in colon tissues using flow cytometry by four markers including an anti-mouse lineage cocktail (FITC anti-mCD3/FITC anti-mGr-1/FITC anti-mCD11b/ FITC anti-mCD45R (B220)/FITC anti-mTer-119), PE/Cy7 anti-mouse CD45, PE anti-mouse CD117 (c-kit), and APC anti-mouse IL-33 Rα (ST2). Results: The total ILC population was significantly higher in the chemically induced reparative change compared with the sham group. ILC1 percentage in the chemically induced reparative change was significantly higher compared to those in the other three groups (Sham, chemically induced dysplasia and orthotopic dysplasia). The orthotopic dysplasia group showed more ILC3 percentage than the other groups. Conclusion: ILC1 and ILC3 subgroups increased significantly in reparative and dysplastic experimental CRC respectively. Thus ILC1 may have an inhibitory effect on tumor growth whereas ILC3 promotes tumor progression.
Ziba Veisi Malekshahi; Nasser Hashemi Goradel; Mehdi Shakouri Khomartash; Amir Maleksabet; Maryam kadkhodazadeh; Gholam Ali Kardar; Babak Negahdari
Abstract
Background: Human colorectal cancer cells overexpress carcinoembryonic antigen (CEA). CEA is a glycoprotein which has shown to be a promising vaccine target for immunotherapy against colorectal cancer. Objective: To design a DNA vaccine harboring CEA antigen and evaluate its effect on inducing immunity ...
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Background: Human colorectal cancer cells overexpress carcinoembryonic antigen (CEA). CEA is a glycoprotein which has shown to be a promising vaccine target for immunotherapy against colorectal cancer. Objective: To design a DNA vaccine harboring CEA antigen and evaluate its effect on inducing immunity against colorectal cancer cells in tumor bearing mice. Methods: In the first step the coding sequence of the CEA was cloned into the pcDNA3.1 vector. The mice were injected with the vaccine construct and the immune responses were monitored during the experiment period. The specific IgG anti-CEA, IFN-γ, IL-2 and IL-4 were measured by ELISA and levels of IFN-γ was detected by ELISpot assay. The lymphocyte proliferation was assessed using a 5-bromo-2-deoxyuridine (BrdU) cell proliferation assay kit. Results: Immunization of the mice with the CEA plasmid resulted in stimulation of CEA-specific T cell and antibody responses. The serum level of specific IgG antibodies against CEA was increased in immunized mice. Moreover, the injection of CEA plasmid led to the stimulation of T-helper-1 by increase in the secretion of IFN-γ, IL-2 and lymphocyte proliferation response. Conclusion: As the CEA DNA vaccine displayed encouraging antitumor effects, therefore, we suggest that it can be a potential therapeutic modality for colorectal cancer and is worthy of further investigation.
Morteza Akbari; Dariush Shanehbandi; Milad Asadi; Navid Shomali; Afsaneh Faraji; Vahid Khaze; Abbas Pakdel; Ahad Mokhtarzadeh; Ali Asghar Ebrahimi; Aliakbar Shabani; Behzad Baradaran
Abstract
Background: Colorectal cancer (CRC) is attributed as one of the most common malignancies worldwide. CD133 molecule, as a pentaspan transmembrane glycoprotein, confers stem cell-related characteristics, including self-renewal and multi-directional differentiation capability. CD133 plays important roles ...
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Background: Colorectal cancer (CRC) is attributed as one of the most common malignancies worldwide. CD133 molecule, as a pentaspan transmembrane glycoprotein, confers stem cell-related characteristics, including self-renewal and multi-directional differentiation capability. CD133 plays important roles in the progression of CRC by conferring apoptotic resistance and migration ability. Objective: To investigate the anti-apoptotic and anti-angiogenic effect of CD-133 targeted siRNA in a colorectal cancer cell line. Methods: In this study, CD133-targeted siRNA transfection was conducted into HT-29 cells. MTT assay was employed to evaluate the cytotoxic effects of transfection on the cells. Flow cytometry was used to evaluate the apoptosis rate. The mRNA expression of apoptosis and metastasis related genes were assessed by quantitative Real-Time PCR (qRT-PCR). Wound healing assay was used to assess the migration potency of the infected cells. Results: Expression of CD133 was significantly downregulated after transfection of CD133-specific siRNA. Moreover, the rate of apoptosis was significantly increased after transfection. The migration potential of cells was diminished after transfection. siRNA delivery resulted in the modulation of expression of apoptosis and metastasis-related genes. Conclusion: siRNA mediated targeting of CD133 could be considered as a promising approach to treat CRC through suppressing the cancerous behavior of tumor cells.
