Oxidative Stress and DNA Damage in Mice Kidneys Exposed to Cadmium Chloride

Background: Cadmium is one of the most toxic heavy metals in our environments having a very strong ability to accumulate in body organs especially in kidney. Our aim of this study is to determine of the genotoxicity and cytotoxicity in mice kidneys exposed to cadmium. Material and Method: In this study we sacrificed 30 male mice and randomly divided into 2 different groups (control& case). Every 5 mice remained in one cage at the same standard conditions. After a week, the mice were exposed to Cd (300µm/Kg.b.wt) on 0, 6, 12, 24, 48 hrs by peritoneal injections. After 24hrs from the latest injection, the mice were killed and obtained their kidneys, and then the oxidative stress markers (malondialdehide (MDA), glutathione (GSH) and superoxide dismutase (SOD)) were assayed on homogenized kidneys in order to study of cytotoxicity, genotoxicity and DNA damage. We used Comet assay on separated kidney cells. Finally, for analyzing statistical data, we used T-test and ANOVA using SPSS15 software. Results: In treated group, MDA and GSH concentration and also SOD (ρ<0.05) activity significantly increased in comparison with control group. Conclusion: The comet assay results obviously showed DNA breakage stimulated by Cd in treated group it was not seen in the control group

Increased Expression of CCAT2 LncRNA in Non-Melanoma Skin Cancer

Background: The non-melanoma skin cancer is one of the most prevalent type of skin cancers, which at least involves 2-3 million people annually. In recent decades, we have witnessed a considerable rise in the incidence of NMSC in Iran. In this paper, we studied the expression of the new lncRNA colon cancer-associated transcript 2 (CCAT2) in cases of non-melanoma skin cancer (NMSC).Methods: The sample included 36 patients and 30 healthy subjects, of whom, we extracted the total RNA from tissues. Using the cDNA synthase, we conducted the real time PCR. Using the SPSS software, we analyzed the data and drew the graphs by PRISM software. The index of P<0.05 was considered significant.Results: The values of CCAT2, TCF7L2 and MYC indicated a considerable expression rise in the NMSCs in comparison with the controls. In addition, the expression of CCAT2 was found  to be higher in high-grade tumors than low-grade tumors. According to results, there is a relationship between CCAT2 and NMSC initiation as well as the progression. The CCAT2 functions by its downstream genes, TCF7L2 and MYC, with an impact on the Wnt signaling pathway.Conclusions: based on the results, the lncRNA CCAT2 acts as a potential biomarker for NMSC pathogenesis

Increased Expression of CCAT2 LncRNA in Non-Melanoma Skin Cancer

Background: The non-melanoma skin cancer is one of the most prevalent type of skin cancers, which at least involves 2-3 million people annually. In recent decades, we have witnessed a considerable rise in the incidence of NMSC in Iran. In this paper, we studied the expression of the new lncRNA colon cancer-associated transcript 2 (CCAT2) in cases of non-melanoma skin cancer (NMSC).Methods: The sample included 36 patients and 30 healthy subjects, of whom, we extracted the total RNA from tissues. Using the cDNA synthase, we conducted the real time PCR. Using the SPSS software, we analyzed the data and drew the graphs by PRISM software. The index of P<0.05 was considered significant.Results: The values of CCAT2, TCF7L2 and MYC indicated a considerable expression rise in the NMSCs in comparison with the controls. In addition, the expression of CCAT2 was found  to be higher in high-grade tumors than low-grade tumors. According to results, there is a relationship between CCAT2 and NMSC initiation as well as the progression. The CCAT2 functions by its downstream genes, TCF7L2 and MYC, with an impact on the Wnt signaling pathway.Conclusions: based on the results, the lncRNA CCAT2 acts as a potential biomarker for NMSC pathogenesis

Inhibition of AGS cancer cell proliferation following siRNA-mediated downregulation of VEGFR2

Vascular endothelial growth factor (VEGF) and VEGF receptors (VEGFRs) play important roles in angiogenesis of different developmental mechanisms such as wound healing, embryogenesis and diseases, including different types of cancer. VEGFR2 is associated with cell proliferation, migration, and vascular permeability of endothelial cells. Blocking VEGF and its receptors is suggested as a therapeutic approach to prevent tumor growth. In this study, we aim to block VEGF signaling via small interfering RNA (siRNA) inhibition of VEGFR2. Materials and Methods: In this experimental study, we used the RNA interference (RNAi) mechanism to suppress expression of the VEGFR2 gene. We conducted the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide (MTT) assay, real-time polymerase chain reaction (PCR), Western blot, and?ow cytometry analyses of VEGFR2 expression. Results: Real-time PCR and Western blot results showed that VEGFR2 expression signifcantly downregulated. This suppression was followed by inhibition of cell proliferation, reduction of viability, and induction of apoptosis in the cancer cells. Conclusion: These findings suggest that VEGFR2 has a role in cell proliferation and tumor growth. Accordingly, it is suggested that VEGFR2 can be a therapeutic target for controlling tumor growth and proliferation

Effect of Acute Toxicity of Cadmium in Mice Kidney Cells

Background: Cadmium is one of the most toxic heavy metals in our environment having a very strong ability to accumulate in body organs, especially in kidney. The present study was done to determine the genotoxicity and cytotoxicity in kidneys of rats exposed to cadmium. Methods: Male rats (n=30), kept in standard conditions were used in this study. The animals were randomly divided into 2 groups (control and treatment). The treatment group was intraperitoneally injected with Cd (300µm/kg) at hours 0, 6, 12, 24, 48. Twenty four hours after the last injection, the rats were sacrificed and their kidneys were obtained. Then oxidative stress markers, malondialdehide (MDA), glutathione (GSH), and superoxide dismutase (SOD), were assayed in homogenized kidney for studying their cytotoxicity. For genotoxicity and DNA damage studies, Comet assay was run on isolated kidney cells. Data analysis was done by t-test and ANOVA using SPSS software version 15. Results: MDA and GSH concentrations in normal and Cd exposed kidney cells were 287.01±37.30nmol/g.pr and 15.61±3.89µmol/g.pr and 609.24±87.87nmol/g.pr and 28.52±5.22µmol/g.pr, respectively. In addition, SOD activity in normal and Cd exposed kidney cells were 77.75±4.12 and 218.91±5.40 U/mg.pr, respectively. Comet assay results (content comet length, tail length, and head diameter) showed DNA breakage in the treatment group that was stimulated by Cd which was not seen in the control group. Conclusion: The results demonstrated the genotoxicity effect of Cd on kidney cells as well as the ability of Cd to producing cytotoxicity

Preventing the expression of VEGFR-1 in culture medium using specific SiRNA - as a potential therapeutic method in eye neovascularization

Background and Aim: Angiogenesis is one of important biological processes any disruption in which leads to disease. The main signaling factor in this process is VEGF which acts through its receptors. The present study was done in order to inhibit the expression of receptor type1of this factor (VEGFR-1) using specific siRNA in the culture medium to use its inhibitory effect on neovascularization in the eye. Materials and Methods: In this experimental study first, using target gene sequences, sequences of the specific siRNA were designed against them blasted and manufactured. On the other hand, cDNA of HUVEC cell was synthesized and PCR, with specific primers for target gene, was reproduced as necessary. Then, PEFGP-N1 expression vectors were cloned and confirmed. Then, the obtained plasmid vector was transferred to Hela cells lacking target expressive genes through lipofectamin. GFR expression rate in the initial vector and in the cloned one, both in presence and in absence of specific VEGFR1 siRNA, was assessed. Evaluation of gene inhibition was carried out through decreasing of green fluorescence from GFR, Western blot and RT-PCR. Results were analyzed using T-test and P<0.05 was taken as the significant level. Results: The fluorescence emission from defined siRNA decreased compared to control group. SDS pages and blots from vector cloned cells exposed to both siRNA showed reduced protein expression The outcome of applying two siRNA indicates gene expression in the form of transcription and translation, compared to the control group (P<0.05). Conclusion: Specifically designed siRNA against VEGFR1, through lipofectamin, was appropriately transferred into cell and significantly prevented from the receptor expression. In fact, by blocking angiogenesis signaling route, it was able to prevent neovascurization. Thus, this can be made use of as an appropriate factor in preventing or decreasing neovascularization in the eye