Pub Date : 2019-09-08DOI: 10.22074/cellj.2020.6537
F. Strube, M. Infanger, M. Wehland, Xenia Delvinioti, A. Romswinkel, C. Dietz, A. Kraus
Objective Weightlessness simulation due to the simulated microgravity has been shown to considerably affect behavior of tumor cells. It is aim of this study to evaluate characteristics of human breast cancer cells in this scaffold- free 3D culture model. Materials and Methods In this experimental study, the cells were exposed to simulated microgravity in a random- positioning machine (RPM) for five days. Morphology was observed under phase-contrast and confocal microscopy. Cytofilament staining was performed and changes in expression level of cytofilament genes, proliferation/differentiation genes, oncogenes and tumor suppressor genes were detected by quantitative reverse transcription polymerase chain reaction (qRT-PCR), followed by western blot confirmation. Results After five days, distinct spheroid formation was observed. Rearrangement of the cytoskeleton into spherical shape was visible. VIM gene expression was significantly up-regulated for adherent cells and spheroids (3.3x and 3.6x respectively, P<0.05 each). RHOA also showed significant gene up-regulation for adherent cells and spheroids (3.2x and 3.9x respectively, P<0.05 each). BRCA showed significant gene up-regulation in adherent cells and spheroids (2.1x and 4.1x respectively, P<0.05 each). ERBB2 showed significant gene up-regulation (2.4x, P<0.05) in the spheroids, but not in the adherent cells. RAB27A showed no significant alteration in gene expression. MAPK) showed significant gene up-regulation in adherent cells and spheroids (3.2x, 3.0x, P<0.05 each). VEGF gene expression was down-regulated under simulated microgravity, without significance. Alterations of gene expressions could be confirmed on protein level for vimentin and MAPK1. Protein production was not increased for BRCA1, human epidermal growth factor receptor 2 (HER2) and VEGF. Contradictory changes were determined for RHOA and its related protein. Conclusion Microgravity provides an easy-to handle, scaffold-free 3D-culture model for human breast cancer cells. There were considerable changes in morphology, cytoskeleton shape and gene expressions. Identification of the underlying mechanisms could provide new therapeutic options.
{"title":"Alteration of Cytoskeleton Morphology and Gene Expression in Human Breast Cancer Cells under Simulated Microgravity","authors":"F. Strube, M. Infanger, M. Wehland, Xenia Delvinioti, A. Romswinkel, C. Dietz, A. Kraus","doi":"10.22074/cellj.2020.6537","DOIUrl":"https://doi.org/10.22074/cellj.2020.6537","url":null,"abstract":"Objective Weightlessness simulation due to the simulated microgravity has been shown to considerably affect behavior of tumor cells. It is aim of this study to evaluate characteristics of human breast cancer cells in this scaffold- free 3D culture model. Materials and Methods In this experimental study, the cells were exposed to simulated microgravity in a random- positioning machine (RPM) for five days. Morphology was observed under phase-contrast and confocal microscopy. Cytofilament staining was performed and changes in expression level of cytofilament genes, proliferation/differentiation genes, oncogenes and tumor suppressor genes were detected by quantitative reverse transcription polymerase chain reaction (qRT-PCR), followed by western blot confirmation. Results After five days, distinct spheroid formation was observed. Rearrangement of the cytoskeleton into spherical shape was visible. VIM gene expression was significantly up-regulated for adherent cells and spheroids (3.3x and 3.6x respectively, P<0.05 each). RHOA also showed significant gene up-regulation for adherent cells and spheroids (3.2x and 3.9x respectively, P<0.05 each). BRCA showed significant gene up-regulation in adherent cells and spheroids (2.1x and 4.1x respectively, P<0.05 each). ERBB2 showed significant gene up-regulation (2.4x, P<0.05) in the spheroids, but not in the adherent cells. RAB27A showed no significant alteration in gene expression. MAPK) showed significant gene up-regulation in adherent cells and spheroids (3.2x, 3.0x, P<0.05 each). VEGF gene expression was down-regulated under simulated microgravity, without significance. Alterations of gene expressions could be confirmed on protein level for vimentin and MAPK1. Protein production was not increased for BRCA1, human epidermal growth factor receptor 2 (HER2) and VEGF. Contradictory changes were determined for RHOA and its related protein. Conclusion Microgravity provides an easy-to handle, scaffold-free 3D-culture model for human breast cancer cells. There were considerable changes in morphology, cytoskeleton shape and gene expressions. Identification of the underlying mechanisms could provide new therapeutic options.","PeriodicalId":9692,"journal":{"name":"Cell Journal (Yakhteh)","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2019-09-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"90448909","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2019-09-08DOI: 10.22074/cellj.2020.6548
Fariba Azimipour, S. Zavareh, Taghi Lashkarbolouki
Objective The unfavorable effects of electromagnetic radiation (EMR) emitted by the cell phone on reproduction health are controversial. Metalloproteinases play a vital role in ovarian follicle development. This study was designed to investigate the effects of exposure to the cell phone on the gelatinolytic activity of in vitro cultured mouse pre-antral follicle. Materials and Methods In this experimental study, pre-antral follicles were isolated from ovaries of immature mice (n=16) and cultured with or without exposure to the cell phone in talking mode for 60 minutes. The gelatinolytic activity was evaluated through the zymography method, as well as the gene expression of matrix metalloproteinases (MMPs) namely MMP-2 and -9 and tissue inhibitors of metalloproteinases (TIMPs) namely, TIMP-1 and -2 by the real-time polymerase chain reaction (PCR) method. Also, in parallel, the development of pre-antral follicles was assessed. Results The maturation parameters of the cell phone-exposed pre-antral follicles were significantly lower compared with the control group (P<0.05). The gelatinolytic activity was significantly decreased in the cell phone-exposed pre- antral follicles compared with the control group (P<0.05). The relative mRNA expression of the MMP-2 gene was significantly (P<0.05) increased in the cell phone-exposed pre-antral follicles whereas the expression rate of the MMP-9 gene was considerably (P<0.05) reduced when compared with the control group. Conversely, the relative expression of the TIMP-1 was markedly (P<0.05) increased in the cell phone-exposed pre-antral follicles while the expression of the TIMP-2 was (P<0.05) significantly diminished in comparison with the control group. Conclusion Exposure to the cell phone alters the growth and maturation rate of murine ovarian follicle through the changing in the expression of the MMP-2 and -9 genes, as well as the gelatinolytic activity.
{"title":"The Effect of Radiation Emitted by Cell Phone on The Gelatinolytic Activity of Matrix Metalloproteinase-2 and -9 of Mouse Pre-Antral Follicles during In Vitro Culture","authors":"Fariba Azimipour, S. Zavareh, Taghi Lashkarbolouki","doi":"10.22074/cellj.2020.6548","DOIUrl":"https://doi.org/10.22074/cellj.2020.6548","url":null,"abstract":"Objective The unfavorable effects of electromagnetic radiation (EMR) emitted by the cell phone on reproduction health are controversial. Metalloproteinases play a vital role in ovarian follicle development. This study was designed to investigate the effects of exposure to the cell phone on the gelatinolytic activity of in vitro cultured mouse pre-antral follicle. Materials and Methods In this experimental study, pre-antral follicles were isolated from ovaries of immature mice (n=16) and cultured with or without exposure to the cell phone in talking mode for 60 minutes. The gelatinolytic activity was evaluated through the zymography method, as well as the gene expression of matrix metalloproteinases (MMPs) namely MMP-2 and -9 and tissue inhibitors of metalloproteinases (TIMPs) namely, TIMP-1 and -2 by the real-time polymerase chain reaction (PCR) method. Also, in parallel, the development of pre-antral follicles was assessed. Results The maturation parameters of the cell phone-exposed pre-antral follicles were significantly lower compared with the control group (P<0.05). The gelatinolytic activity was significantly decreased in the cell phone-exposed pre- antral follicles compared with the control group (P<0.05). The relative mRNA expression of the MMP-2 gene was significantly (P<0.05) increased in the cell phone-exposed pre-antral follicles whereas the expression rate of the MMP-9 gene was considerably (P<0.05) reduced when compared with the control group. Conversely, the relative expression of the TIMP-1 was markedly (P<0.05) increased in the cell phone-exposed pre-antral follicles while the expression of the TIMP-2 was (P<0.05) significantly diminished in comparison with the control group. Conclusion Exposure to the cell phone alters the growth and maturation rate of murine ovarian follicle through the changing in the expression of the MMP-2 and -9 genes, as well as the gelatinolytic activity.","PeriodicalId":9692,"journal":{"name":"Cell Journal (Yakhteh)","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2019-09-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"82027356","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2019-09-08DOI: 10.22074/cellj.2020.6340
Amir Bajouri, Z. Orouji, E. Taghiabadi, A. Nazari, Atefeh Shahbazi, N. Fallah, P. Mohammadi, Mohammad Rezvani, Zahra Jouyandeh, Fatemeh Vaezirad, Zahra Khalajasadi, M. Ghasemi, Aslan Fanni, Sara Haji Hosseinali, A. Alizadeh, H. Baharvand, Saeed Shafieyan, N. Aghdami
Objective Recently, the promising potential of fibroblast transplantation has become a novel modality for skin rejuvenation. We investigated the long-term safety and efficacy of autologous fibroblast transplantation for participants with mild to severe facial contour deformities. Materials and Methods In this open-label, single-arm phase IIa clinical trial, a total of 57 participants with wrinkles (n=37, 132 treatment sites) or acne scars (n=20, 36 treatment sites) who had an evaluator’s assessment score of at least 2 out 7 (based on a standard photo-guide scoring) received 3 injections of autologous cultured fibroblasts administered at 4-6 week intervals. Efficacy evaluations were performed at 2, 6, 12, and 24 months after the final injection based on evaluator and patient’s assessment scores. Results Our study showed a mean improvement of 2 scores in the wrinkle and acne scar treatment sites. At sixth months after transplantation, 90.1% of the wrinkle sites and 86.1% of the acne scar sites showed at least a one grade improvement on evaluator assessments. We also observed at least a 2-grade improvement in 56.1% of the wrinkle sites and 63.9% of the acne scar sites. A total of 70.5% of wrinkle sites and 72.2% of acne scar sites were scored as good or excellent on patient assessments. The efficacy outcomes remained stable up to 24-month. We did not observe any serious adverse events during the study. Conclusion These results have shown that autologous fibroblast transplantation could be a promising remodeling modality with long-term corrective ability and minimal adverse events (Registration Number: NCT01115634).
{"title":"Long-Term Follow-up of Autologous Fibroblast Transplantation for Facial Contour Deformities, A Non-Randomized Phase IIa Clinical Trial","authors":"Amir Bajouri, Z. Orouji, E. Taghiabadi, A. Nazari, Atefeh Shahbazi, N. Fallah, P. Mohammadi, Mohammad Rezvani, Zahra Jouyandeh, Fatemeh Vaezirad, Zahra Khalajasadi, M. Ghasemi, Aslan Fanni, Sara Haji Hosseinali, A. Alizadeh, H. Baharvand, Saeed Shafieyan, N. Aghdami","doi":"10.22074/cellj.2020.6340","DOIUrl":"https://doi.org/10.22074/cellj.2020.6340","url":null,"abstract":"Objective Recently, the promising potential of fibroblast transplantation has become a novel modality for skin rejuvenation. We investigated the long-term safety and efficacy of autologous fibroblast transplantation for participants with mild to severe facial contour deformities. Materials and Methods In this open-label, single-arm phase IIa clinical trial, a total of 57 participants with wrinkles (n=37, 132 treatment sites) or acne scars (n=20, 36 treatment sites) who had an evaluator’s assessment score of at least 2 out 7 (based on a standard photo-guide scoring) received 3 injections of autologous cultured fibroblasts administered at 4-6 week intervals. Efficacy evaluations were performed at 2, 6, 12, and 24 months after the final injection based on evaluator and patient’s assessment scores. Results Our study showed a mean improvement of 2 scores in the wrinkle and acne scar treatment sites. At sixth months after transplantation, 90.1% of the wrinkle sites and 86.1% of the acne scar sites showed at least a one grade improvement on evaluator assessments. We also observed at least a 2-grade improvement in 56.1% of the wrinkle sites and 63.9% of the acne scar sites. A total of 70.5% of wrinkle sites and 72.2% of acne scar sites were scored as good or excellent on patient assessments. The efficacy outcomes remained stable up to 24-month. We did not observe any serious adverse events during the study. Conclusion These results have shown that autologous fibroblast transplantation could be a promising remodeling modality with long-term corrective ability and minimal adverse events (Registration Number: NCT01115634).","PeriodicalId":9692,"journal":{"name":"Cell Journal (Yakhteh)","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2019-09-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"80272507","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2019-09-08DOI: 10.22074/cellj.2020.6550
Nasrin Zare, Shaghayegh Haghjooy Javanmard, V. Mehrzad, N. Eskandari, A. Andalib
Objective The purpose of this study was to investigate effect of plasma-derived exosomes of refractory/relapsed or responsive diffuse large B-cell lymphoma (DLBCL) patients on natural killer (NK) cell functions. Materials and Methods In this cross-sectional and experimental study, NK cells were purified from responsive patients (n=10) or refractory/relapsed patients (n=12) and healthy donors (n=12). NK cells were treated with plasma-derived exosomes of responsive or refractory/relapsed patients. We examined the expression levels of hsa-miR-155-5p, hsa- let-7g-5p, INPP5D (SHIP-1) and SOCS-1 in NK cells quantitative reverse transcription-polymerase chain reaction (qRT-PCR). Percentages of NK cells expressing CD69, NKG2D and CD16, NK cell cytotoxicity and NK cell proliferation (using flow-cytometry) as well as interferon-gamma (IFN-γ) level in the supernatant of NK cells using ELISA were also investigated. Results We observed an increased level of hsa-miR-155-5p and a decreased level of SOCS-1 in NK cells treated with exosomes compared to untreated NK cell in healthy donors and DLBCL patients. An increase in hsa-miR-155-5p level was associated with an increased level of IFN-γ in healthy donors. The decreased levels of hsa-let-7g-5p were observed in NK cells treated with exosomes in comparison with untreated NK cells in DLBCL patients (P<0.05). There was no significant difference in the percentage of CD69+NK cells and NKG2D+ NK cells in the absence or presence of exosomes of DLBCL patients in each group. Furthermore, we observed significant reduction of NK cell proliferation in DLBCL patients and healthy donors in the presence of exosomes of refractory/relapsed patients (P<0.05). A significant decrease was observed in cytotoxicity of NK cell in patients with DLBCL treated with exosomes of responsive patients. Conclusion Our findings demonstrated adverse effect of plasma-derived exosomes of DLBCL patients on some functions of NK cell. It was also determined that low NK cell count might be associated with impaired response to R-CHOP and an increased recurrence risk of cancer.
目的探讨难治性/复发性或反应性弥漫性大b细胞淋巴瘤(DLBCL)患者血浆源性外泌体对自然杀伤细胞(NK)功能的影响。材料和方法在这项横断面和实验研究中，从反应性患者(n=10)或难治性/复发患者(n=12)和健康供者(n=12)中纯化NK细胞。NK细胞用反应性或难治性/复发患者的血浆来源外泌体处理。我们检测了hsa- mir -155-5p、hsa- let-7g-5p、INPP5D (SHIP-1)和SOCS-1在NK细胞中的表达水平。用ELISA法检测NK细胞上清液中表达CD69、NKG2D和CD16的NK细胞百分比、NK细胞毒性和NK细胞增殖(流式细胞术)以及干扰素γ (IFN-γ)水平。结果我们观察到，与健康供体和DLBCL患者的NK细胞相比，外泌体处理的NK细胞中hsa-miR-155-5p水平升高，SOCS-1水平降低。健康供体中hsa-miR-155-5p水平升高与IFN-γ水平升高相关。与未处理NK细胞相比，外泌体处理NK细胞中hsa-let-7g-5p水平降低(P<0.05)。两组DLBCL患者外泌体存在或缺失时CD69+NK细胞和NKG2D+ NK细胞的百分比差异无统计学意义。此外，我们观察到在难治性/复发患者的外泌体存在时，DLBCL患者和健康供者的NK细胞增殖显著减少(P<0.05)。反应性患者外泌体治疗的DLBCL患者NK细胞毒性显著降低。结论DLBCL患者血浆源性外泌体对NK细胞的某些功能有不良影响。研究还确定，低NK细胞计数可能与R-CHOP反应受损和癌症复发风险增加有关。
{"title":"Effect of Plasma-Derived Exosomes of Refractory/Relapsed or Responsive Patients with Diffuse Large B-Cell Lymphoma on Natural Killer Cells Functions","authors":"Nasrin Zare, Shaghayegh Haghjooy Javanmard, V. Mehrzad, N. Eskandari, A. Andalib","doi":"10.22074/cellj.2020.6550","DOIUrl":"https://doi.org/10.22074/cellj.2020.6550","url":null,"abstract":"Objective The purpose of this study was to investigate effect of plasma-derived exosomes of refractory/relapsed or responsive diffuse large B-cell lymphoma (DLBCL) patients on natural killer (NK) cell functions. Materials and Methods In this cross-sectional and experimental study, NK cells were purified from responsive patients (n=10) or refractory/relapsed patients (n=12) and healthy donors (n=12). NK cells were treated with plasma-derived exosomes of responsive or refractory/relapsed patients. We examined the expression levels of hsa-miR-155-5p, hsa- let-7g-5p, INPP5D (SHIP-1) and SOCS-1 in NK cells quantitative reverse transcription-polymerase chain reaction (qRT-PCR). Percentages of NK cells expressing CD69, NKG2D and CD16, NK cell cytotoxicity and NK cell proliferation (using flow-cytometry) as well as interferon-gamma (IFN-γ) level in the supernatant of NK cells using ELISA were also investigated. Results We observed an increased level of hsa-miR-155-5p and a decreased level of SOCS-1 in NK cells treated with exosomes compared to untreated NK cell in healthy donors and DLBCL patients. An increase in hsa-miR-155-5p level was associated with an increased level of IFN-γ in healthy donors. The decreased levels of hsa-let-7g-5p were observed in NK cells treated with exosomes in comparison with untreated NK cells in DLBCL patients (P<0.05). There was no significant difference in the percentage of CD69+NK cells and NKG2D+ NK cells in the absence or presence of exosomes of DLBCL patients in each group. Furthermore, we observed significant reduction of NK cell proliferation in DLBCL patients and healthy donors in the presence of exosomes of refractory/relapsed patients (P<0.05). A significant decrease was observed in cytotoxicity of NK cell in patients with DLBCL treated with exosomes of responsive patients. Conclusion Our findings demonstrated adverse effect of plasma-derived exosomes of DLBCL patients on some functions of NK cell. It was also determined that low NK cell count might be associated with impaired response to R-CHOP and an increased recurrence risk of cancer.","PeriodicalId":9692,"journal":{"name":"Cell Journal (Yakhteh)","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2019-09-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"75129750","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2019-09-08DOI: 10.22074/cellj.2020.6486
M. Fattahi, N. Eskandari, F. Sotoodehnejadnematalahi, V. Shaygannejad, K. Mohammad
Objective Multiple sclerosis (MS) is an inflammatory disease resulting in demyelination of the central nervous system (CNS). T helper 17 (Th17) subset protects the human body against pathogens and induces neuroinflammation, which leads to neurodegeneration. MicroRNAs (miRNAs) are a specific class of small (~22 nt) non-coding RNAs that act as post-transcriptional regulators. The expression of the miR-326 is highly associated with the pathogenesis of MS disease in patients through the promotion of Th17 development. Recently, studies showed that disease-modifying therapies (DMTs) could balance the dysregulation of miRNAs in the immune cells of patients with relapsing-remitting MS (RRMS). Interferon-beta (IFN-β) has emerged as one of the most common drugs for the treatment of RR-MS patients. The purpose of this study was to evaluate the expression of the miR-326 in RRMS patients who were responders and non- responders to IFN-β treatment. Materials and Methods In this cross-sectional study, a total of 70 patients (35 responders and 35 non-responders) were enrolled. We analyzed the expression of the miR-326 in peripheral blood mononuclear cells (PBMCs) of RRMS patients at least one year after the initiation of IFN-β therapy. Real-time polymerase chain reaction (RT-PCR) was applied to measure the expression of the miR-326. Results The results showed no substantial change in the expression of the miR-326 between responders and non- responders concerning the treatment with IFN-β. Although the expression of the miR-326 was slightly reduced in IFN-β-responders compared with IFN-β-non-responders; however, the reduction of the miR-326 was not statistically significant. Conclusion Overall, since IFN-β doesn’t normalize abnormal expression of miR-326, this might suggest that IFN-β affects Th17 development through epigenetic mechanisms other than miR-326 regulation.
{"title":"Comparison of The Expression of miR-326 between Interferon beta Responders and Non-Responders in Relapsing-Remitting Multiple Sclerosis","authors":"M. Fattahi, N. Eskandari, F. Sotoodehnejadnematalahi, V. Shaygannejad, K. Mohammad","doi":"10.22074/cellj.2020.6486","DOIUrl":"https://doi.org/10.22074/cellj.2020.6486","url":null,"abstract":"Objective Multiple sclerosis (MS) is an inflammatory disease resulting in demyelination of the central nervous system (CNS). T helper 17 (Th17) subset protects the human body against pathogens and induces neuroinflammation, which leads to neurodegeneration. MicroRNAs (miRNAs) are a specific class of small (~22 nt) non-coding RNAs that act as post-transcriptional regulators. The expression of the miR-326 is highly associated with the pathogenesis of MS disease in patients through the promotion of Th17 development. Recently, studies showed that disease-modifying therapies (DMTs) could balance the dysregulation of miRNAs in the immune cells of patients with relapsing-remitting MS (RRMS). Interferon-beta (IFN-β) has emerged as one of the most common drugs for the treatment of RR-MS patients. The purpose of this study was to evaluate the expression of the miR-326 in RRMS patients who were responders and non- responders to IFN-β treatment. Materials and Methods In this cross-sectional study, a total of 70 patients (35 responders and 35 non-responders) were enrolled. We analyzed the expression of the miR-326 in peripheral blood mononuclear cells (PBMCs) of RRMS patients at least one year after the initiation of IFN-β therapy. Real-time polymerase chain reaction (RT-PCR) was applied to measure the expression of the miR-326. Results The results showed no substantial change in the expression of the miR-326 between responders and non- responders concerning the treatment with IFN-β. Although the expression of the miR-326 was slightly reduced in IFN-β-responders compared with IFN-β-non-responders; however, the reduction of the miR-326 was not statistically significant. Conclusion Overall, since IFN-β doesn’t normalize abnormal expression of miR-326, this might suggest that IFN-β affects Th17 development through epigenetic mechanisms other than miR-326 regulation.","PeriodicalId":9692,"journal":{"name":"Cell Journal (Yakhteh)","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2019-09-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"78007559","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2019-09-08DOI: 10.22074/cellj.2020.6492
Javid Sabour Takanlu, Arad Aghaie Fard, Saeed Mohammdi, S. M. A. Hosseini Rad, S. Abroun, M. Nikbakht
Objective Multiple myeloma (MM) is an incurable plasma cell malignancy. Several genetic and epigenetic changes affect numerous critical genes expression status in this disorder. CDKN2A gene is expressed at low level in almost all cases with MM disease. The mechanism of this gene down-regulation has remained controversial. In the present study, we targeted EZH2 by microRNA-124 (miR-124) in L-363 cells and assessed following possible impact on CDKN2A gene expression and phenotypic changes. Materials and Methods In this experimental study, growth inhibitory effects of miR-124 were measured by MTT assay in L-363 cell line. Likewise, cell cycle assay was measured by flowcytometery. The expression levels of EZH2 and CDKN2A were evaluated by real-time quantitative reverse-transcription polymerase chain reaction (qRT-PCR). Results qRT-PCR results showed induction of EZH2 gene expression after transduction of cells with lentivector expressing miR-124. The expression of CDKN2A was also upregulated as the result of EZH2 supression. Coincide with gene expression changes, cell cycle analysis by flow-cytometry indicated slightly increased G1-arrest in miR- transduced cells (P<0.05). MTT assay results also showed a significant decrease in viability and proliferation of miR- transduced cells (P<0.05). Conclusion It seems that assembling of H3K27me3 mark mediated by EZH2 is one of the key mechanisms of suppressing CDKN2A gene expression in MM disease. However, this suppressive function is applied by a multi-factor mechanism. In other words, targeting EZH2, as the core functional subunit of PRC2 complex, can increase expression of the downstream suppressive genes. Consequently, by increasing expression of tumor suppressor genes, myeloma cells are stopped from aberrant expansions and they become susceptible to regulated cellular death.
{"title":"Indirect Tumor Inhibitory Effects of MicroRNA-124 through Targeting EZH2 in The Multiple Myeloma Cell Line","authors":"Javid Sabour Takanlu, Arad Aghaie Fard, Saeed Mohammdi, S. M. A. Hosseini Rad, S. Abroun, M. Nikbakht","doi":"10.22074/cellj.2020.6492","DOIUrl":"https://doi.org/10.22074/cellj.2020.6492","url":null,"abstract":"Objective Multiple myeloma (MM) is an incurable plasma cell malignancy. Several genetic and epigenetic changes affect numerous critical genes expression status in this disorder. CDKN2A gene is expressed at low level in almost all cases with MM disease. The mechanism of this gene down-regulation has remained controversial. In the present study, we targeted EZH2 by microRNA-124 (miR-124) in L-363 cells and assessed following possible impact on CDKN2A gene expression and phenotypic changes. Materials and Methods In this experimental study, growth inhibitory effects of miR-124 were measured by MTT assay in L-363 cell line. Likewise, cell cycle assay was measured by flowcytometery. The expression levels of EZH2 and CDKN2A were evaluated by real-time quantitative reverse-transcription polymerase chain reaction (qRT-PCR). Results qRT-PCR results showed induction of EZH2 gene expression after transduction of cells with lentivector expressing miR-124. The expression of CDKN2A was also upregulated as the result of EZH2 supression. Coincide with gene expression changes, cell cycle analysis by flow-cytometry indicated slightly increased G1-arrest in miR- transduced cells (P<0.05). MTT assay results also showed a significant decrease in viability and proliferation of miR- transduced cells (P<0.05). Conclusion It seems that assembling of H3K27me3 mark mediated by EZH2 is one of the key mechanisms of suppressing CDKN2A gene expression in MM disease. However, this suppressive function is applied by a multi-factor mechanism. In other words, targeting EZH2, as the core functional subunit of PRC2 complex, can increase expression of the downstream suppressive genes. Consequently, by increasing expression of tumor suppressor genes, myeloma cells are stopped from aberrant expansions and they become susceptible to regulated cellular death.","PeriodicalId":9692,"journal":{"name":"Cell Journal (Yakhteh)","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2019-09-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"78799365","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2019-09-08DOI: 10.22074/cellj.2020.6525
Nazanin Karimi, H. Mohseni Kouchesfahani, M. Nasr-Esfahani, M. Tavalaee, A. Shahverdi, H. Choobineh
Objective The aim of this blind randomised clinical trial study was to assess the clinical efficiency of combined density gradient centrifugation/Zeta (DGC/Zeta) sperm selection procedure compared to conventional DGC in infertile men candidates for intracytoplasmic sperm injection (ICSI). The literature shows that DGC/Zeta is more effective compared to DGC alone in selection of sperms with normal chromatin and improves the clinical outcome of the ICSI procedure. Therefore, this study re-evaluates the efficiency of DGC/Zeta in improving the clinical outcomes of ICSI in an independent clinical setting. Materials and Methods In this randomized, single-blind, clinical trial, a total of 240 couples with male factor infertility and at least one abnormal sperm parameter were informed regarding the study and 220 participated. Based on inclusion and exclusion criteria, 103 and 102 couples were randomly allocated into the DGC/Zeta and DGC groups, respectively. ICSI outcomes were followed and compared between the two groups. Results Although there was no significant difference in fertilization rate (P=0.67) between the DGC/Zeta and DGC groups, mean percentage of good embryo quality (P=0.04), good blastocysts quality (P=0.049), expanded blastocysts (P=0.007), chemical pregnancies (P=0.005) and clinical pregnancies (P=0.007) were significantly higher in the DGC/ Zeta group compared to DGC. In addition, implantation rate was insignificantly higher in DGC/Zeta compared to DGC (P=0.17). Conclusion This is the second independent study showing combined DGC/Zeta procedure improves ICSI outcomes, especially the pregnancy rate, compared to the classical DGC procedure and this is likely related to the improved quality of sperm selected by the DGC/Zeta procedure (Registration number: IRCT20180628040270N1).
{"title":"DGC/Zeta as A New Strategy to Improve Clinical Outcome in Male Factor Infertility Patients following Intracytoplasmic Sperm Injection: A Randomized, Single-Blind, Clinical Trial","authors":"Nazanin Karimi, H. Mohseni Kouchesfahani, M. Nasr-Esfahani, M. Tavalaee, A. Shahverdi, H. Choobineh","doi":"10.22074/cellj.2020.6525","DOIUrl":"https://doi.org/10.22074/cellj.2020.6525","url":null,"abstract":"Objective The aim of this blind randomised clinical trial study was to assess the clinical efficiency of combined density gradient centrifugation/Zeta (DGC/Zeta) sperm selection procedure compared to conventional DGC in infertile men candidates for intracytoplasmic sperm injection (ICSI). The literature shows that DGC/Zeta is more effective compared to DGC alone in selection of sperms with normal chromatin and improves the clinical outcome of the ICSI procedure. Therefore, this study re-evaluates the efficiency of DGC/Zeta in improving the clinical outcomes of ICSI in an independent clinical setting. Materials and Methods In this randomized, single-blind, clinical trial, a total of 240 couples with male factor infertility and at least one abnormal sperm parameter were informed regarding the study and 220 participated. Based on inclusion and exclusion criteria, 103 and 102 couples were randomly allocated into the DGC/Zeta and DGC groups, respectively. ICSI outcomes were followed and compared between the two groups. Results Although there was no significant difference in fertilization rate (P=0.67) between the DGC/Zeta and DGC groups, mean percentage of good embryo quality (P=0.04), good blastocysts quality (P=0.049), expanded blastocysts (P=0.007), chemical pregnancies (P=0.005) and clinical pregnancies (P=0.007) were significantly higher in the DGC/ Zeta group compared to DGC. In addition, implantation rate was insignificantly higher in DGC/Zeta compared to DGC (P=0.17). Conclusion This is the second independent study showing combined DGC/Zeta procedure improves ICSI outcomes, especially the pregnancy rate, compared to the classical DGC procedure and this is likely related to the improved quality of sperm selected by the DGC/Zeta procedure (Registration number: IRCT20180628040270N1).","PeriodicalId":9692,"journal":{"name":"Cell Journal (Yakhteh)","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2019-09-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"73887245","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2019-09-08DOI: 10.22074/cellj.2020.6549
Keivan Mobini, Elham Banakar, G. Tamaddon, A. Mohammadi-Bardbori
Objective microRNAs (miRNAs) play bifunctional roles in the initiation and progression of cancer, and recent evidence has confirmed that unusual expression of miRNAs is required for the progress of breast cancer. The regulatory role of aryl hydrocarbon receptor (AhR) and its endogenous ligand, 6-formylindolo[3,2-b]carbazole (FICZ) on the expression of tumor suppressor miRNAs, miR-22, miR-515-5p and miR-124-3p, as well as their association with the estrogen receptor alpha (ERα) were the aims of this study. Materials and Methods In this experimental study, the expression levels of miR-22, miR-515-5p, miR-124-3p and miR-382-5p in MCF-7 cells were determined using the quantificational real time polymerase chain reaction (qRT-PCR) assay. Results Our results revealed that miR-22, miR-515-5p, and miR-124-3p expressions were significantly increased in cells transfected with ERα siRNA. Our data also showed that miR-22, miR 515-5p, and miR-124-3p expression levels were significantly increased following FICZ treatment. Here, we found that AhR/ERα cross-talk plays a critical role in the expression of miR-22, miR-515-5p and miR-124-3p in MCF-7 cells. Conclusion Overall, our data demonstrated that FICZ, as an AhR agonist could induce the expression of tumor suppressor miRNAs, miR-22, miR-515-5p, and miR-124-3p; thus, FICZ might be regarded as a potential therapeutic agent for breast cancer treatment.
{"title":"6-Formylindolo[3,2-b]carbazole (FICZ) Enhances The Expression of Tumor Suppressor miRNAs, miR-22, miR-515-5p, and miR-124-3p in MCF-7 Cells","authors":"Keivan Mobini, Elham Banakar, G. Tamaddon, A. Mohammadi-Bardbori","doi":"10.22074/cellj.2020.6549","DOIUrl":"https://doi.org/10.22074/cellj.2020.6549","url":null,"abstract":"Objective microRNAs (miRNAs) play bifunctional roles in the initiation and progression of cancer, and recent evidence has confirmed that unusual expression of miRNAs is required for the progress of breast cancer. The regulatory role of aryl hydrocarbon receptor (AhR) and its endogenous ligand, 6-formylindolo[3,2-b]carbazole (FICZ) on the expression of tumor suppressor miRNAs, miR-22, miR-515-5p and miR-124-3p, as well as their association with the estrogen receptor alpha (ERα) were the aims of this study. Materials and Methods In this experimental study, the expression levels of miR-22, miR-515-5p, miR-124-3p and miR-382-5p in MCF-7 cells were determined using the quantificational real time polymerase chain reaction (qRT-PCR) assay. Results Our results revealed that miR-22, miR-515-5p, and miR-124-3p expressions were significantly increased in cells transfected with ERα siRNA. Our data also showed that miR-22, miR 515-5p, and miR-124-3p expression levels were significantly increased following FICZ treatment. Here, we found that AhR/ERα cross-talk plays a critical role in the expression of miR-22, miR-515-5p and miR-124-3p in MCF-7 cells. Conclusion Overall, our data demonstrated that FICZ, as an AhR agonist could induce the expression of tumor suppressor miRNAs, miR-22, miR-515-5p, and miR-124-3p; thus, FICZ might be regarded as a potential therapeutic agent for breast cancer treatment.","PeriodicalId":9692,"journal":{"name":"Cell Journal (Yakhteh)","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2019-09-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"80464948","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2019-09-08DOI: 10.22074/cellj.2020.6302
Zahraa Nasheed Hamad Almohammed, Fatemeh Moghani-Ghoroghi, I. ragerdi-Kashani, R. Fathi, Leila Sadat Tahaei, Mohamad Naji, P. Pasbakhsh
Objective This study examined the in vitro effect of melatonin on the protein synthesis of mitochondria, as well as autophagy in matured oocytes of aged mice. Materials and Methods In this experimental study, germinal vesicles (GV) oocytes were collected from aged (with the age of six-months-old) and young mice (with age range of 6-8 weeks old) and then cultured in the in vitro culture medium (IVM) for 24 hours to each metaphase II (MII) oocytes and then supplemented with melatonin at a concentration of 10 μM. The culture medium of MII oocytes was devoid of melatonin. Afterward, the expression of the SIRT-1 and LC3 was assessed by immunocytochemistry. ATP-dependent luciferin-luciferase bioluminescence assay was employed for the measurement of the ATP contents. Intracellular reactive oxygen specious (ROS) was detected by DCFH-DA, and the total antioxidant capacity (TAC) level was determined by TAC assay. Results The expression of SIRT-1 and LC3, as well as the measurement of the ATP content, was significantly increased in oocytes treated with melatonin compared with the oocytes receiving no treatment. Moreover, TAC was considerably higher in melatonin-treated oocytes than oocytes receiving no treatment. On the other hand, the level of ROS was significantly decreased in oocytes treated with melatonin in comparison with the untreated oocytes. The results indicated that melatonin considerably improved the development of oocytes as well. Conclusion According to the data, melatonin increased mitochondrial function and autophagy via an increase in the expression of SIRT1 and LC3, as well as the ATP contents while it decreased the levels of ROS and increased TAC in oocytes derived from aged mice.
{"title":"The Effect of Melatonin on Mitochondrial Function and Autophagy in In Vitro Matured Oocytes of Aged Mice","authors":"Zahraa Nasheed Hamad Almohammed, Fatemeh Moghani-Ghoroghi, I. ragerdi-Kashani, R. Fathi, Leila Sadat Tahaei, Mohamad Naji, P. Pasbakhsh","doi":"10.22074/cellj.2020.6302","DOIUrl":"https://doi.org/10.22074/cellj.2020.6302","url":null,"abstract":"Objective This study examined the in vitro effect of melatonin on the protein synthesis of mitochondria, as well as autophagy in matured oocytes of aged mice. Materials and Methods In this experimental study, germinal vesicles (GV) oocytes were collected from aged (with the age of six-months-old) and young mice (with age range of 6-8 weeks old) and then cultured in the in vitro culture medium (IVM) for 24 hours to each metaphase II (MII) oocytes and then supplemented with melatonin at a concentration of 10 μM. The culture medium of MII oocytes was devoid of melatonin. Afterward, the expression of the SIRT-1 and LC3 was assessed by immunocytochemistry. ATP-dependent luciferin-luciferase bioluminescence assay was employed for the measurement of the ATP contents. Intracellular reactive oxygen specious (ROS) was detected by DCFH-DA, and the total antioxidant capacity (TAC) level was determined by TAC assay. Results The expression of SIRT-1 and LC3, as well as the measurement of the ATP content, was significantly increased in oocytes treated with melatonin compared with the oocytes receiving no treatment. Moreover, TAC was considerably higher in melatonin-treated oocytes than oocytes receiving no treatment. On the other hand, the level of ROS was significantly decreased in oocytes treated with melatonin in comparison with the untreated oocytes. The results indicated that melatonin considerably improved the development of oocytes as well. Conclusion According to the data, melatonin increased mitochondrial function and autophagy via an increase in the expression of SIRT1 and LC3, as well as the ATP contents while it decreased the levels of ROS and increased TAC in oocytes derived from aged mice.","PeriodicalId":9692,"journal":{"name":"Cell Journal (Yakhteh)","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2019-09-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"86432144","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2019-07-31DOI: 10.22074/cellj.2020.6158
S. Peymanfar, R. Roghanian, K. Ghaedi, Sayed-Hamid Zarkesh-Esfahani, R. Yari
Objective Granulocyte colony-stimulating factor (G-CSF) has a wide variety of functions including stimulation of hematopoiesis and proliferation of granulocyte progenitor cells. Recombinant human G-CSF (rh-G-CSF) is used for treatment of neutropenia in patients receiving chemotherapy. The mature bloodstream neutrophils express G-CSF receptor (G-CSFR), presenting a significant and specific mechanism for circulating G-CSF clearance. Computational studies are essential bioinformatics methods used for characterization of proteins with regard to their physicochemical properties and 3D configuration, as well as protein–ligand interactions for recombinant drugs. We formerly produced rh-G-CSF in E. coli and showed that the isolated protein had unacceptable biological activity in mice. In the present paper, we aimed to characterize the purified rh-G-CSF by analytical tests and developed an in vivo model by computational modelling of G-CSF. Materials and Methods In this experimental study, we analyzed the purified G-CSF using the analytical experiments. Then, the crystalline structure was extracted from Protein Data Bank (PDB) and molecular dynamics (MD) simulation was performed using Gromacs 5.1 package under an Amber force field. The importance of amino acid contents of G-CSF, to bind the respective receptor was also detected; moreover, the effect of dithiothreitol (DTT) used in G-CSF purification was studied. Results The results revealed that characteristics of the produced recombinant G-CSF were comparable with those of the standard G-CSF and the recombinant G-CSF with the residual amino acid was stable. Also, purification conditions (DTT and existence of extra cysteine) had a significant effect on the stability and functionality of the produced G-CSF. Conclusion Experimental and in silico analyses provided good information regarding the function and characteristics of our recombinant G-CSF which could be useful for industrial researches.
目的粒细胞集落刺激因子(G-CSF)具有多种功能，包括刺激造血和粒细胞祖细胞增殖。重组人G-CSF (rh-G-CSF)用于治疗化疗患者的中性粒细胞减少症。成熟的中性粒细胞表达G-CSF受体(G-CSFR)，在循环G-CSF清除中具有重要的特异性机制。计算研究是基本的生物信息学方法，用于描述蛋白质的物理化学性质和三维构型，以及重组药物的蛋白质-配体相互作用。我们以前在大肠杆菌中产生了rh-G-CSF，并表明分离的蛋白质在小鼠中具有不可接受的生物活性。在本文中，我们旨在通过分析测试来表征纯化的rh-G-CSF，并通过G-CSF的计算建模建立了体内模型。材料与方法本实验对纯化后的G-CSF进行分析实验。然后，从蛋白质数据库(Protein Data Bank, PDB)中提取晶体结构，并在Amber力场下使用Gromacs 5.1软件包进行分子动力学(MD)模拟。还检测了G-CSF的氨基酸含量对结合相应受体的重要性;此外，还研究了二硫苏糖醇(DTT)在G-CSF纯化中的作用。结果制备的重组G-CSF与标准G-CSF具有相当的特性，且重组G-CSF中氨基酸残留稳定。此外，纯化条件(DTT和多余半胱氨酸的存在)对生成的G-CSF的稳定性和功能有显著影响。结论重组G-CSF的实验分析和计算机分析为其功能和特性提供了良好的信息，可用于工业研究。
{"title":"Characterization and In Silico Analysis of The Structural Features of G-CSF Derived from Lysates of Escherichia coli","authors":"S. Peymanfar, R. Roghanian, K. Ghaedi, Sayed-Hamid Zarkesh-Esfahani, R. Yari","doi":"10.22074/cellj.2020.6158","DOIUrl":"https://doi.org/10.22074/cellj.2020.6158","url":null,"abstract":"Objective Granulocyte colony-stimulating factor (G-CSF) has a wide variety of functions including stimulation of hematopoiesis and proliferation of granulocyte progenitor cells. Recombinant human G-CSF (rh-G-CSF) is used for treatment of neutropenia in patients receiving chemotherapy. The mature bloodstream neutrophils express G-CSF receptor (G-CSFR), presenting a significant and specific mechanism for circulating G-CSF clearance. Computational studies are essential bioinformatics methods used for characterization of proteins with regard to their physicochemical properties and 3D configuration, as well as protein–ligand interactions for recombinant drugs. We formerly produced rh-G-CSF in E. coli and showed that the isolated protein had unacceptable biological activity in mice. In the present paper, we aimed to characterize the purified rh-G-CSF by analytical tests and developed an in vivo model by computational modelling of G-CSF. Materials and Methods In this experimental study, we analyzed the purified G-CSF using the analytical experiments. Then, the crystalline structure was extracted from Protein Data Bank (PDB) and molecular dynamics (MD) simulation was performed using Gromacs 5.1 package under an Amber force field. The importance of amino acid contents of G-CSF, to bind the respective receptor was also detected; moreover, the effect of dithiothreitol (DTT) used in G-CSF purification was studied. Results The results revealed that characteristics of the produced recombinant G-CSF were comparable with those of the standard G-CSF and the recombinant G-CSF with the residual amino acid was stable. Also, purification conditions (DTT and existence of extra cysteine) had a significant effect on the stability and functionality of the produced G-CSF. Conclusion Experimental and in silico analyses provided good information regarding the function and characteristics of our recombinant G-CSF which could be useful for industrial researches.","PeriodicalId":9692,"journal":{"name":"Cell Journal (Yakhteh)","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2019-07-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"83127689","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
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