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.
Lia Farahi; Fatemeh Ghaemimanesh; Saeideh Milani; Seyed Mohsen Razavi; Reza Hadavi; Ali Ahmad Bayat; Ali Salimi; Mohammad Mehdi Akhondi; Hodjattallah Rabbani
Abstract
Background: We have previously reported the aberrant expression of Fibromodulin (FMOD) in patients with chronic lymphocytic leukemia (CLL). Although FMOD has been considered as a cytoplasmic or secretory protein, we discovered the cell surface expression of FMOD in leukemic B cells via anchoring with ...
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Background: We have previously reported the aberrant expression of Fibromodulin (FMOD) in patients with chronic lymphocytic leukemia (CLL). Although FMOD has been considered as a cytoplasmic or secretory protein, we discovered the cell surface expression of FMOD in leukemic B cells via anchoring with glycosylphosphatidylinositol (GPI). Objective: To evaluate FMOD as a new biomarker in CLL patients in comparison with healthy individuals. Methods: A monoclonal antibody was generated against human FMOD. The cell surface expression of FMOD in 52 CLL patients and 45 healthy individuals were compared by flow cytometry. A bacterial phosphatidylinositol-specific phospholipase C (PI-PLC) was used to determine the cell surface localization of FMOD using ELISA and flow cytometry techniques. Annexin V-FITC and propidium iodide (PI) was used to detect apoptosis induction in CLL PBMCs following in vitro incubation with anti-FMOD mAb. Results: The results demonstrated the widespread cell surface expression of GPI-anchored FMOD in CLL patients (median: 79.9 %), although healthy individuals had low FMOD expression (median: 6.2 %) (p≤0.0001). The cut-off value of FMOD expression was estimated with high sensitivity and specificity at 17.9%. Furthermore, in vitro apoptosis induction of leukemic cells following incubation with anti-FMOD mAb showed a direct apoptosis of CLL cells (27.9%) with very low effect on healthy PBMCs (6%). Conclusion: The membrane-anchoring of FMOD by means of a GPI moiety in leukemic cells supports FMOD as a highly potential diagnostic and therapeutic target in CLL patients.
Cheah Wen Yapp; Amal Widaad Mohaimin; Adi Idris
Volume 14, Issue 3 , September 2017, , Pages 250-256
Abstract
Background: Cytosolic double-stranded RNA (dsRNA) is an important ‘molecular signature’ for the detection of intracellular viral infections. Although intracellular dsRNA is a known potent inducer of apoptosis, the optimal time and dose for the onset of dsRNA-mediated apoptosis have not been ...
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Background: Cytosolic double-stranded RNA (dsRNA) is an important ‘molecular signature’ for the detection of intracellular viral infections. Although intracellular dsRNA is a known potent inducer of apoptosis, the optimal time and dose for the onset of dsRNA-mediated apoptosis have not been studied in detail. Objective: To perform an accurate temporal assessment of the cell death responses in dsRNA-dependent cytotoxicity. Methods: Poly I:C (PIC), a synthetic dsRNA molecule was delivered intracellularly into J774.1 and RAW264.7 murine macrophages via electroporation. Cell viability was measured using the MTT assay and apoptosis was determined by sub-G0/G1 DNA content using flow cytometry. Results: Loss of cell viability was seen as early as 3h post-electroporation of macrophages. A significant increase in the sub-G0/G1 DNA content consistent with apoptosis was observed in PIC-electroporated macrophages as early as 3h post electroporation. Conclusion: Intracellular PIC delivery induces rapid macrophage cell death.
Lida Toomarian; Mandana Sattari; Nazanin Hashemi; Nikoo Tadayon; Alireza Akbarzadeh Baghban
Abstract
Background: The infectious nature of severe early-childhood caries (S-ECC) points to the possible participation of immunologic host responses including neutrophils and their antimicrobial products. Objectives: The aim of this study was to determine the neutrophil apoptosis, α-defensins (HNP1-3) ...
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Background: The infectious nature of severe early-childhood caries (S-ECC) points to the possible participation of immunologic host responses including neutrophils and their antimicrobial products. Objectives: The aim of this study was to determine the neutrophil apoptosis, α-defensins (HNP1-3) and calprotectin levels in the saliva of preschool children and the association with S-ECC. Methods: Oral examinations were performed on 87 children aged 3-5 years and non stimulated whole saliva samples were collected. Thirty of these subjects were considered S-ECC children, 30 with moderate caries (MC) and 27 were caries free (CF). To detect apoptosis, cell staining was done with Annexin-V-Fluos and propidium iodide, and they were analyzed by fluorescent microscopy. The concentration of α-defensins and calprotectin were assessed using ELISA. Results: There were no statistical differences between groups considering the HNP1-3 or calprotectin salivary levels (p=0.06 and p=0.23, respectively). The HNP1-3 and calprotectin levels were negatively correlated and the correlation was significant in MC group (p=0.03). Lower levels of apoptotic neutrophils were obtained from CF subjects as compared with S-ECC children (p=0.03). Conclusions: Our findings establish that apoptotic mechanisms could be implicated in the immunity responses associated with S-ECC. We cannot yet
Mohammadreza Ataollahi; Mansour Salehi; Iman Doostan; Zahra Kabiri; Mohammadreza Mohajeri; Farzaneh Mahmoodi; Raheleh Shokouhi; Shadi Javan; Mohammad Hassan Meshkibaf; Behnoosh Miladpoor
Abstract
Background: Apoptosis and cell cycle regulation play an important role in pathogenesis and tumor progression in patients with Diffuse Large B-Cell Lymphoma (DLBCL). Bcl-2 associated athanogene-1 (BAG-1) is an antiapoptotic protein as well as a regulator of cell growth. There is no conclusive evidence ...
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Background: Apoptosis and cell cycle regulation play an important role in pathogenesis and tumor progression in patients with Diffuse Large B-Cell Lymphoma (DLBCL). Bcl-2 associated athanogene-1 (BAG-1) is an antiapoptotic protein as well as a regulator of cell growth. There is no conclusive evidence about BAG-1 protein expression in this disease. Objective: To investigate the expression level of BAG-1 protein in DLBCL. Methods: Thirty patients diagnosed from 1997-2004, as having DLBCL, were selected. Also 30 normal lymph nodes were included as normal counterparts in this study. BAG-1 expression was determined by immunohistochemical staining in both DLBCL and normal lymph node samples. Results: Of the 30 DLBCLs examined, 100% were positive for nuclear and 83% were positive for cytoplasmic BAG-1 staining. Of the 30 normal lymph nodes investigated, 20% were positive for nuclear and 0% were positive for cytoplasmic BAG-1 staining. Nuclear staining in DLBCL samples was significantly higher than those of normal lymph nodes (100% versus 20%, p <0.001). Besides, cytoplasmic staining in DLBCL samples was significantly higher than those of normal lymph nodes (83% versus 0%, p <0.001). There was no association between BAG-1 staining and patients' overall survival. Conclusion: Our data indicated that BAG-1 protein was deregulated in this disease similar to some other malignancies such as breast and colon cancer. Overexpression of BAG-1 in DLBCL suggests that this protein probably plays an important role in the pathogenesis of DLBCL. Besides, higher nuclear BAG-1 staining might be correlated with poor prognosis.
Mohammad Reza Razeghinejad; Eskandar Kamali-Sarvestani
Volume 4, Issue 4 , December 2007, , Pages 215-219
Abstract
Background: Glaucoma is one of the most common causes of blindness and is usually associated with elevated intraocular pressure. In patients with primary open angle glau-coma the number of trabecular meshwork cells is decreased. Death of the trabecular meshwork cells may be a result of apoptosis. Objective: ...
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Background: Glaucoma is one of the most common causes of blindness and is usually associated with elevated intraocular pressure. In patients with primary open angle glau-coma the number of trabecular meshwork cells is decreased. Death of the trabecular meshwork cells may be a result of apoptosis. Objective: To investigate the aqueous humor levels of soluble Fas (sFas) and Fas-Ligand (sFasL) in glaucomatous patients. Methods: Concentration of sFas and sFasL were measured by ELISA in 41 eyes with glaucoma (21 with pseudoexfoliation and 20 with primary open angle glaucoma) and 39 eyes with cataract as controls. Results: The sFas concentration was lower in the pri-mary open angle than the pseudoexfoliation glaucoma and the cataract groups (p=0.002 and p= 0.004, respectively). The sFasL level did not show any significant difference in the three groups. Conclusion: A lower level of sFas may provide proper microenvironment for increased apoptosis of trabecular meshwork cells in primary open angle glaucoma.
Nasser Gholijani; Ahmad Monabati; Zahra Amirghofran
Volume 1, Issue 1 , June 2004, , Pages 34-40
Abstract
Background: Prostate cancer is one of the most commonly diagnosed cancers in males. Tumor suppressor gene p53 plays an important role in causing cell cycle arrest and allowing apoptosis to proceed. Objective: To investigate the expression of p53 protein and its relation to apoptosis and prostate cancer ...
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Background: Prostate cancer is one of the most commonly diagnosed cancers in males. Tumor suppressor gene p53 plays an important role in causing cell cycle arrest and allowing apoptosis to proceed. Objective: To investigate the expression of p53 protein and its relation to apoptosis and prostate cancer traditional prognostic indicators. Methods: In this study expression of p53 was examined in paraffin-embedded tissues from 50 cases of prostate carcinoma by immunohistochemistry and evaluated using an index of staining. Correlation between p53 expression and apoptosis was detected by TUNEL method. Pathological grade, Gleason score and stage of carcinoma were also determined. Results: P53 expression was observed in 48 of 50 cases (26-100% of tumor cells) with mean staining index of 141±65. A significant association between p53 expression and pathologic grade (r=0.37, p=0.004) and Gleason score (r= 0.4, p=0.009) of patients was observed. Apoptosis was detected in only 6 patients. p53 expression showed no correlation with apoptotic index. No correlation between p53 expression and stage or apoptosis and clinicopathological characteristics of patients was found. Conclusion: p53 expression showed a significant correlation with differentiation status of the prostate carcinoma and can be helpful as a prognostic marker. Decreased level of apoptosis observed in our cases was not correlated with p53 expression indicating the possible role of other regulatory molecules involved in the apoptosis.
