International Journal of Molecular and Cellular Medicine最新文献

The NF-kB signaling pathway was introduced as a key pathway in carcinogenesis that is induced by inflammation in gastrointestinal malignancies. The RelA transcription factor is an important component of this signaling pathway. Furthermore, CD44 is implicated in the tumorigenesis and metastasis of gastric cancer. The aim of this study was to assay the effect of RELA knockout on CD44 expression in MKN45 cells. CRISPR/Cas9 was used to knock out RELA in MKN-45. The median fluorescence intensity (MFI) of CD44 before and after RELA knockout is analyzed in MKN45. The CRISPR/Cas9 vector pSpCas9 (BB)-2A-Puro (PX459) was used for gRNA cloning (two guides). The MKN-45 cell line was co-transfected. The purified co-transfected cells with puromycin were cultured and used for the RELA gene expression assay by real-time PCR. Flow cytometry was used for the analysis of the MFI of CD44+ in MKN45. The results showed that 180 nucleotide sequences between exon 2 and exon 3 of RELA were deleted in MKN45. RELA expression significantly (P<0.001) decreased after CRISPR/Cas9 knockout. Compared to the control group, the MFI of CD44 in transfected cells significantly decreased (P <0.001). Knockout of RELA significantly decreased CD44 expression in MKN45 cells. It can be concluded that the NF-kB signaling pathway via RELA is related to CD44 expression and consequently the tumorigenesis of gastric cancer. More studies about this relationship are recommended.

Sulforaphane (SFN) is an organosulfur product of found isothiocyanates in vegetables. The chemopreventive effects of SFN have revealed that there is a link between excessive consumption of SFN-rich vegetables and cancer formation without possible toxicological consequences. We aimed to evaluate the cellular outcome of SFN from a toxicological perspective, particularly for renal cells including clear cell adenocarcinoma (769-P) and human embryonic renal epithelial (293T) cells. The viability/cytotoxicity experiments were performed with methyl thiazole diphenyl tetrazolium (MTT) and lactate dehydrogenase (LDH) assays. IC₅₀-dependent, non-cytotoxic concentrations were used for the determination of cell cycle status and apoptosis by using flow cytometry and western blot. A certain concentration of SFN effectively altered apoptotic/necrotic behavior in 769-P compared to the control group 293T. Cell cycle status remained stable while showing a decreased proliferation profile for 769-P cells. The percentage of the S phase from the cell cycle in 293T cells significantly reduced without affecting proliferation status. The use of SFN as an alternative to traditional treatments might be considered for the battle against renal cell carcinoma but the current findings showed that caution should be applied particularly for renal cells. Our study will provide a basis for future in vivo studies to support traditional cancer therapies.

Capsaicin is a natural product which is extracted from pepper and has the potential to be used in cancer treatment because of its anti- proliferative effects. The aim of the study was to determine the effect of capsaicin on the hepatocellular carcinoma cell proliferation and the expressions of related genetic markers as Ki-67, PI3K/AKT/mTOR and epigenetic markers as miR-126 and piR-Hep-1. The inhibitory concentration of capsaicin in HepG2 cells was determined. piR-Hep-1 and miR-126 expressions and Ki-67, PI3K, AKT and mTOR gene expressions were examined by RT-PCR. The inhibitory concentration of capsaicin for HepG2 cells was 200 nM and the decreased proliferation was observed at 24^th hour. As epigenetic markers, an up regulation of miR-126 and down regulation of piR-Hep-1 expression were determined after treatment. Moreover, Ki-67, PI3K and mTOR gene expressions decreased while AKT gene expression increased after the treatment (p<0.001). According to the obtained data, capsaicin has an impact on proliferation both genetically and epigenetically. Furthermore, treatment of capsaicin effects miR-126 and piR-Hep-1 expressions which effect carcinogenesis in different way. Moreover, there are some clues which indicate that these two small non-coding RNA might affect each other and share the same target molecules post-transcriptionally.

The aim of the present study was to examine the hypothesis that miR-33-5p attenuates morphine state-dependent (StD) memory via the µ opioid receptor by regulating cyclic AMP response element-binding protein (CREB). The effects of post-training morphine and morphine StD memory and their interaction with pre-test naloxone were evaluated using a single-trial inhibitory avoidance paradigm. Then, the hippocampal miR-33-5p gene and pCREB/CREB protein expression profiles were evaluated using quantitative real-time PCR and western blotting, respectively. We found that while post-training morphine and morphine StD memory respectively up- and down-regulate the miR-33-5p expression profile in the hippocampus, the reverse results are true for the expression of pCREB/CREB. Pre-test naloxone antagonized the response. Overall, our findings suggest that the expression levels of miR-33-5p in the hippocampus set the basis for morphine StD memory with low miR-33-5p enabling state dependency. The mechanism is mediated via miR33-5p and CREB signaling with the interaction of the µ opioid receptor. This finding may be used as a potential strategy for ameliorating morphine-induced memory-related disorders.

Caveolin-1(Cav-1) is one of the most important components of caveolae in the cell membrane, which plays an important role in cell signaling transduction, such as EGFR and TGF-β receptor transactivation. The purpose of this study was to evaluate the effect of c-Abl and NAD(P)H oxidases (NOX) on phosphorylation of Cav-1 and consequently their effect on phosphorylation of Smad2C induced by Endothelin-1 in human vascular smooth muscle cells (VSMCs). In this study, all experiments were performed using human VSMCs. The phosphorylation level of the Caveolin-1 and Smad2C proteins were assessed by western blotting using Phospho-Caveolin-1 (Tyr14) antibody and phospho-Smad2 (Ser465/467) antibody. The data were reported as mean ± SEM. The VSMCs treated with endothelin-1(ET-1) (100 nanomolar (nmol)) demonstrated a time-dependent increase in the pCav-1 level (p<0.05). The inhibitors of NOX (diphenyleneiodonium) (p<0.05), cholesterol depleting agent (beta-cyclodextrin) (p<0.05) and c-Abl inhibitor (PP1) (p<0.01) were able to reduce the level of the phospho-Cav-1 and phospho-Smad2C induced by Et-1 (p<0.05). Our results proposed that caveolae structure, NOX, c-Abl played an important role in the phosphorylation of Cav-1 induced by ET-1 in the human VSMCs. Furthermore, our findings showed that phosphoCav-1 involved in TGFR transactivation. Thus, Et-1 via a transactivation-dependent mechanism can cause phosphorylation of Smad2C.

The aim of the present study was to investigate the expression and methylation pattern of the angiotensin I converting enzyme 2 (ACE-2) in acute respiratory distress syndrome (ARDS) covid-9 patients. A total of 25 patients with covid-19 ARDS and 20 controls were recruited. Expression of the ACE-2 gene was evaluated by quantitative real time PCR, and methylation of CpG dinucleotides, in the ACE-2 promoter, was quantified using bisulfite pyro-sequencing. Our results showed high expression of the ACE-2 gene in the blood samples of ADRS patients (1.93± 0.67) in comparison to controls (0.62±0.35) (P = 0.03). Correspondingly, in ARDS bronchoalveolar lavage fluid (BALF) samples, there was a high expression of this gene (1.8±0.78) in comparison to controls (0.58±0.2) (p <0.05). Moreover, the methylation rate of the ACE-2 gene in blood samples of ARDS patients was 64.07 ±6.1 in comparison to controls (80.3 ±7.3) (p<0.0001). In BALF samples, there was this pattern too (55.07 ±3.1 vs. 72.35±5.1) (p<0.0001). Finally, a significant correlation was found between expression and methylation in BALF (R= -0.54, P= 0.002) and blood (R= -0.321, P= 0.013) samples. These results indicated that aberrant methylation of the ACE-2 promoter might be associated with high expression of this gene and the occurrence of ARDS in covid-19 patients.

The current study aimed to investigate the effect of a 12-week endurance training (ET) on microRNA-145 (miR-145) changes and Wnt3a and Dab2 cardiomyocytes genes expression of hypercholesterolemic Wistar male rats. Thirty-two male Wistar rats (191.2±19 g, 6-8 weeks age) were randomly assigned into the aerobic exercise-normal nutrition (ANN; n=8), hypercholesterolemic (HCL; n=8), aerobic exercise- hypercholesterolemic (ACL; n=8), and normal nutrition (NN; n=8). Hypercholesterolemia was created by adding 1% cholesterol to the food of the HCL and ACL rats. ET was done five sessions per week on nonconsecutive days for 12 weeks. Twenty-four hours after the last training session, the rats were killed, and the cardiomyocytes were removed. The expression of miR-145, Wnt3a, and Dab2 genes in cardiomyocytes was assessed by real time PCR method. The expression of miR-145 significantly increased in the ANN group in comparison with other groups (P = 0.001). Also, Dab2 gene expression significantly decreased in the ANN group in comparison with ACL (P = 0.001) and HCL (P = 0.001) groups. The results also showed that the Wnt3a in the ANN group was significantly different from NN (p=0.001), ACL, and HCL (p=0.001) groups. It can be concluded that aerobic training and cholesterol-rich foods play an essential regulatory role in the expression of miR-145, Dab2, and Wnt3a genes. However, cholesterol-rich foods appear to play a more significant regulatory role than aerobic exercise training.

The human papillomavirus (HPV) is a highly contagious and prevalent virus that is primarily sexually transmitted. The Gardasil® quadrivalent vaccine, the Cevarix® bivalent vaccine and the Gardasil® 9 nonavalent vaccine were developed to prevent the spread of HPV as well as the incidence of its associated diseases. The aim of this mini-review is to critically analyze the safety and efficacy of both the Gardasil vaccines. A literature search was conducted on ProQuest, MedLine, Science Direct and Scopus databases. More than hundred articles were scanned, and from this, 38 most relevant papers involving human studies across several countries were closely reviewed. The literature deems the Gardasil® HPV vaccines to be safe and efficacious. Due to the novel nature of these vaccines, long-term efficacies, as well as their associated long-term adverse effects, are yet to be confirmed. Of some concern was the finding that a majority of these studies disclosed minor to major involvement with the vaccine manufacturers, and the inhibitory cost of use in developing nations. Gardasil is largely considered safe to use. However, considering that these vaccines are predominantly indicated for children, further comprehensive, impartial, and long-term studies are needed to critically assess safety and efficacy.

Sodium butyrate (NaBu) is a short-chain fatty acid acting as a histone deacetylase inhibitor, and has been shown to be a potential regulator of cancer cell death. This study aimed to evaluate the effect of NaBu on cell cycle control, apoptosis, and expression of some lncRNAs in two human prostate cancer cells (PC-3 and LNCAP). Cell viability was assessed and the appropriate dose was determined using the MTT assay. Real-time PCR technique was also used to evaluate the expression levels of HULC, FOXCUT, and CCAT2 lncRNAs. Apoptosis was diagnosed using annexin V staining, and cell cycle distribution was then assessed using flow cytometry with propidium iodide DNA staining. NaBu induced apoptosis in both prostate cancer cell lines in a dose-dependent manner. The expressions of CCAT2 and HULC lncRNAs genes have significantly decreased in the presence of NaBu (P <0.05) in both PC3 and LNCAP cell lines, in comparison with the control. However, no significant difference was observed in the expression of FOXCUT lncRNAs. Moreover, the results of flow cytometry showed an increase in cell cycle arrest of LNCAP cell line at the sub-G1 stage as compared to the control cells, but no significant difference was observed between the control cells and NaBu-exposed PC-3 cells. In addition, the percentages of early and late apoptotic cells following treatment with NaBu were 80% and 49.63% in LNCAP and PC-3 cells, respectively. Our results suggest that NaBu has a positive effect on the induction of apoptosis and inhibition of cell cycle in PC-3 and LNCAP prostate cancer cells.

Vimentin, an intermediate filament of mesenchymal cells, is upregulated in epithelial-mesenchymal transition (EMT) and has a main role in cancer metastasis. As a new strategy to control metastatic outgrowth, EMT markers are generally inhibited using some drugs or specific siRNA. In this study, AGS gastric cancer cells were treated with metformin and vimentin-specific siRNA (vim-siRNA) for 48 h. The impact of metformin and vim-siRNA on vimentin downregulation in AGS cells were analyzed by quantitative PCR and Western blot. Following treatment with metformin and vim-siRNA, cell motility, migration and invasion abilities of AGS cells were also analyzed. The results showed that inhibition of vimentin due to metformin was comparable with the vim-siRNA. Furthermore, wound-healing and invasion assays showed a significant decrease in migration and invasion of AGS cells following metformin and vim-siRNA treatment. Our finding for the first time indicated that metformin can be an alternative to specific siRNA for inhibition of vimentin expression and migration of AGS cell line. Taken together, our data indicates that the use of metformin might have a priority to siRNA for inhibition of gastric cancer cell behaviors siRNA is more unstable and expensive than metformin, and needs special vehicles and delivery strategies for efficient transfection of cells. Further in vivo studies can reveal metformin's potential in inhibition of EMT and metastasis of cancer cells.