Trace of survivin in cancer

Survivin is one of the most cancer-specific proteins overexpressed in almost all malignancies, but is nearly undetectable in most normal tissues in adults. Functionally, as a member of the inhibitor of apoptosis family, survivin has been shown to inhibit apoptosis and increase proliferation. The antiapoptotic function of survivin seems to be related to its ability to inhibit caspases directly or indirectly. Furthermore, the role of survivin in cell cycle division control is related to its role in the chromosomal passenger complex. Consistent with its determining role in these processes, survivin plays a crucial role in cancer progression and cancer cell resistance to anticancer drugs and ionizing radiation. On the basis of these findings, recently survivin has been investigated intensively as an ideal tumor biomarker. Thus, multiple molecular approaches such as use of the RNA interfering technique, antisense oligonucleotides, ribozyme, and small molecule inhibitors have been used to downregulate survivin regulation and inhibit its biological function consequently. In this review, all these approaches are explained and other compounds that induced apoptosis in different cell lines through survivin inhibition are also reporte

Effect of Embelin on Inhibition of Cell Growth and Induction of Apoptosis in k562 Cell Line

Background & objectives: Cancer is one of the leading causes of morbidity and mortality worldwide. Leukemia is a cancer of blood cells and bone marrow, which is characterized by abnormal growth of white blood cells, known as blasts. Chronic myeloid leukemia is a clonal hematopoietic stem cell disorder that accounts for 15-20 percent of adult leukemia. Embelin, a natural compound found in the fruit of Embeliaribes plant, has low toxicity and potent anticancer properties. Several studies have shown that the anticancer properties of Embelin are due to inhibition of XIAP (X-linked inhibitor of the apoptosis protein) and modulation of NF-kB signaling pathway. The aim of this study was to investigate the effect of Embelin on the growth and apoptosis of K562 cell line. Methods: K562 cells were cultured in RPMI-1640 medium containing 10 % FBS and 1% penicillin. Then, the cells were treated with different concentrations of Embelin (2, 4, 6, 8 μM/ml) for 72 hours. MTT assay was used to determine the viability of cells. Hoechst staining and DNA electrophoresis were used for apoptosis analysis. Result: Based on the results of MTT assay, Embelin inhibited the viability of K562 cells. The results of Hoechst staining showed that DNA fragmentation was increased in the treated cells. DNA electrophoresis analysis revealed that Embelin induced apoptosis. Discussion: As the results showed, Embelin inhibited the cell growth and induced apoptosis in K562 cells time- and dose-dependently. Therefore, Embelin may be a candidate for treatment of chronic myeloid leukemia. &nbsp

Anti-proliferative Effects of Hydralazine on K562 Cell Line (CML)

Introduction: Cancer is the second leading cause of death in the world and among the types of cancer, leukemia is one of the deadliest. Chronic myeloid leukemia (CML) is the best known types of leukemia. Hydralazine is in the cardiovascular medical group, which is considering as inhibitory factor on the proliferation of cancer cells by knowing its inhibitory effects on DNA methyltransferase. The purpose of this study was to evaluate the inhibitory effects of hydralazine on K562 cells proliferation. Methods: First, the K562 cells were cultured. After, the different concentrations of the hydralazine were prepared and the cells were treated for 24, 48 and 72 hours. Next, the inhibitory effects of this drug on K562 cells proliferation was measured by MTT assay. Hoechst (33342) staining and DNA electrophoresis was used to check apoptosis. Data analyses were performed using SPSS software (Version 20) and ANOVA test (P<0.05). Results: MTT assay results showed that hydralazine has the highest cytotoxic activity in 80&mu;M at 72 hours. Drug IC50 was obtained 81 micromolar. And also the results of Hoechst staining and DNA electrophoresis showed that hydralazine was caused apoptosis. Conclusion: In the present study, it was shown that hydralazine induces cells death by apoptosis and this effect was dose-dependent and treat time dependent. It is suggested that more studies will be done&nbsp;&nbsp; on the effects of this drug on animal models and human

The anti-cancer effect of Propranolol in K562 cell line: an in vitro study

Introduction: &Beta;-AR receptors are one of the proteins involved in cancer and stress. The therapeutic activity of &beta;-blockers such as propranolol is attributed to the blockade of &beta;1-adrenergic receptors (ARs). In this study, the effect of propranolol on the viability of K562 cell line was examined. Material and methods: In order to assessment of anti-tumoral effects of propranolol, different concentrations of propranolol were prepared. K562 cells were treated with different concentrations of propranolol, then the percentage of inhibitory effect of propranolol on K562 cell viability at different times (24, 48 and 72 hours) was estimated by MTT assay. Gel electrophoresis of DNA and DAPI staining were used for apoptosis investigation. Statistical comparisons were performed using two-sample t-test, Nominal significance level of each univariate test was 0.05. Results: Propranolol decreased viability of K562 cell line. The inhibitory effect of propranolol is time- and concentration-dependent, thus in higher concentrations and 72 hours after treatment, the maximum inhibitory effect was observed. (P<0.05). As the results showed, Propranolol induces apoptosis in K562 cell line. Conclusions: With respect to the inhibitory effect of propranolol on cell viability and its apoptotic effect on K562 cell line, this drug may be used for cancer therapy

Anti-proliferative and apoptotic effects of Lovastatin on K562 Erthromyloidy cancer cell line

Background and Objective: Lovastatin is a HMG-CoA reductase inhibitor and used for the treatment of hypercholesterolemia. Inhibition of HMG-CoA reductase results in inhibiting the activity of the Ras proto-oncogene that has mutations in most cancers. This study was done to determine the Anti-proliferative and apoptotic effects of Lovastatin on K562 Erthromyloidy cancer cell line. Methods: The K562 Erthromyloidy cancer cell line were cultured and treated with different concentrations of lovastatin. Their antitumor effect on K562 cells were assessed via MTT assay after 72 hours. Hoechst (33342) staining and DNA electrophoresis were used for study of apoptosis. Results: Lovastatin had antitumor effect on K562 Erthromyloidy cancer cell line and this effect increased by incease of time and concentration.The maximum inhibitory effect was 59% in higher concentration (100 µM) and 72 hours after the treatment. Reduced cell growth at 24 and 48 hours after treatment was 24% and 43%, respectively. Lovastatin significantly inhibited K562 cell growth (P<0.05). Conclusion: This study showed that lovastatin has antitumor effect on K562 Erthromyloidy cancer cell line

Isolation and Cloning of mercuric reductase gene (merA) from mercury-resistant bacteria

Introduction: Some of the bacteria having merA gene coding mineral mercury reducing enzyme, has genetic potential of Hg removing via reduction of mineral mercury and transformation of that to gas form and finally bioremediation of polluted area. The aim of this study is the isolation of merA gene from resistance bacteria and cloning of that into suitable expression vector and then the environmental bioremediation by the transformation of bacteria with this vector. Materials and methods: A number of bacteria were collected in contaminated areas with mercury in order to isolate merA genes. Polymerase chain reaction had done on the four bacterial genomes including Klebsiella pneumoniae, Pseudomonas aeruginosa, Serratia marcescens and Escherichia coli using the specific primers in order to detect merA gene. For cloning, the primers containing restriction enzyme sites are used, merA gene was isolated and amplified. The amplified fragments were cloned in the expression vector pET21a+ and via heat shock method were transformed into E. coli TOP10 competent cell. For clustering of genes, Mega software version 4 was used and bioanformatic studies were achieved for predicted enzyme. Results: merA gene with 1686 bp in length was isolated from K pneumoniae and E. coli. Recombinant vectors in transgenic bacteria were confirmed by various methods and finally were confirmed by sequencing. The result of clustering these genes with existence genes in NCBI showed high similarity. Discussion and conclusion: The existence of merA gene in bacteria that adapted to Hg pollution area is because of resistance, so with cloning this gene into suitable expression vector and transformation of susceptible bacteria with this vector ability of resistance to Hg in bacteria for bioremediation could be given

Synergistic induction of apoptosis in a cell model of human leukemia K562 by nitroglycerine and valproic acid

Nitroglycerin (NG), a nitric oxide donor, and valproic acid (VPA), an inhibitor of histone deacetylases, have impressive effects on numerous cancer cell lines. This study intended to evaluate synergistic effects of NG and VPA on cell viability and apoptosis in K562 cells. K562 cells were cultured in RPMI-1640 supplemented with 10 % heat-inactivated FBS. They were treated with different doses of NG, VPA and cisplatin for 24, 48, and 72 h, and MTT assay was performed to analyze cell viability. Also, Peripheral blood mononuclear cells (PBMC) were cultured in RPMI-1640 media and incubated with NG (200 μM), VAP (100 μM), NG+VPA (150 μM) and cisplatin (8 μM) to evaluate cytotoxicity. IC50 of the drugs, when they were applied separately and in combination, were calculated using the COMPUSYN software. DNA electrophoresis, TUNEL assay, and Hoechst staining were performed to investigate apoptosis induction. RT-PCR was used for the evaluation of apoptotic genes expression. The results of the MTT assay showed that cell viability decreased at all applied doses of NG and VPA. It was noticed that the cytotoxic effects of these drugs were dose- and time-dependent. Based on the COMPUSYN output, the combination of the drugs (VPA and NG) in a certain ratio concentration synergistically decreased cell viability. Cisplatin significantly decreased cell viability of PBMCs and K562 cells. Also, the combination drug had cytotoxic effect and significantly reduced viability of K562 cells compared with PBMCs and control cells. In the target cells treated with this combination, Bax and caspase-3 expression increased but Bcl-2 expression decreased. These results suggest that NG, VPA, and their combination decreased cell viability and induced apoptosis via the intrinsic apoptotic pathway. This study suggests that this combination therapy can be considered for further evaluation as an effective chemotherapeutic strategy for patients with chronic myeloid leukemia

Isolation, cloning and analysis of the hexose transporter 6 gene (HXT6) in a native strain of Saccharomyces cerevisiae IBRC-M30069

Introduction: The Saccharomyces cerevisiae yeast is one of the most important microorganisms to produce ethanol. The S. cerevisiae has 20 genes that encode hexose transporter proteins. Among these gene families, seven genes HXT7-HXT1 have important an role in alcohol production. The researchers proved that alcohol fermentation goes up by increasing the expression of these genes which results in increasing ethanol production. Materials and methods: In this research, isolation of HXT6 gene by specific primers via PCR technology was achieved. The amplified fragments were cloned into pGEM-T vector and transformed to Escherichia coli and sequence analysis was carried out. Results: The nucleotide sequence of open reading frame of HXT6 gene revealed a 1713 bp long with a deduced amino acid of 570 residues. The estimated molecular mass and the predicted isoelectric point of the deduced polypeptide were 62.68 kDa and 7.89 respectively. Discussion and conclusion: The deduced protein sequence showed a high similarity to Hxt6p VL31(EGA76254.1) sequences registered in NCBI and also the highest similarity of this gene with HXT7 ( one of the hexose transporter) was observed. This finding shows that this gene (HXT6) and also HXT7 gene resulted from one ancestor gene by mutation in their functional domain during years

The Anti-cancer effects of Celastrol on K562 cell line

Background and Objective: The level of NF-κB factor expression (a transcriptional factor which increases the expression of inflammatory genes) is often increased in various human cancers. Therefore, NF-κB inhibitors such as Celastrol may prevent cancer development. The purpose of this study was to evaluate the anticancer effects of Celastrol on K562 cells proliferation. Materials and Methods: First, the K562 cells were cultured and cytotoxicity effects of celastrol were determined by MTT assay. Hoechst staining and DNA electrophoresis are used to check apoptosis. Data analysis was performed using SPSS, version 16 and ANOVA test (P<0.05) Results: Statistical analysis of MTT assay data showed that the growth of treated cells with different concentrations of the Celastrol significantly decreased (P<0.05) and inhibitory effect of Celastrol was time and concentration–dependent; so in higher concentrations (8 µM) and 72 hours, the maximum effect has occurred. The IC50 value of Celastrol was obtained 4 µM. Also, the results of Hoechst staining and DNA electrophoresis showed that Celastrol caused fragmentation of cell nucleus and DNA. Conclusion: Based on the results, Celastrol decreases cells viability (P<0.05) and induces apoptosis in K562 cells, its effect is time and dose-dependent. In conclusion, the agent may be applied as an anticancer drug for treatment of chronic myeloid leukemia