Drug resistance mechanisms in a high grade glioma cell line

Glioblastoma multiforme (GBM) is the most common and aggressive primary brain tumour. Despite advances in GBM treatment, there is a high frequency of local relapse due to the acquisition of drug resistance. Investigation of glioma cell lines will help us to understand the molecular basis of this hard to treat tumour. In this study, the rat C6 glioma cell line was used as a model alongside two drug selected derivatives (C6-etoposide and C6-irinotecan) to investigate the mechanisms of chemo-resistance in glioma by identifying candidate proteins, genes, and key signalling pathways. Proteomic (2D gel electrophoresis) and genomic (gene array) analyses were performed to determine protein and gene expression changes. Integration of this data with cellular pathway analysis resulted in the prediction that cellular migration and the response to oxidative stress would be distinct in the drug selected C6 cell lines. Cell migration was subsequently assessed using wound scratch repair and transwell migration assays, whilst the response to oxidative stress produced by reactive oxygen species was determined fluorimetrically. The C6 cell line exposed to irinotecan (DNA topoisomerase I inhibitor) showed reduced migration, even under the influence of chemoattractant, compared to other cell lines, consistent with alterations in the expression of collagen genes. The C6 cell line exposed to etoposide (DNA topoisomerase II inhibitor) showed greater resistance to oxidative stress which was proposed to be due to alterations in the signalling pathways downstream of the PTEN/PI3Kinase. Future studies, investigating the effect of PI3Kinase pathway inhibitors are considered and it is proposed that further research into this signalling pathway will be able to uncover the molecular basis of distinct chemo-resistance in this important model cell system for aggressive glioma

Long-term exposure to irinotecan reduces cell migration in glioma cells.

In spite of considerable research into the therapies for glioblastoma multiforme this tumour type remains very difficult to treat. As well as having a tendency to be inherently resistant to chemotherapy, glioblastoma multiforme also displays local invasion. Cell line studies have a continued and important role to play in understanding the mechanisms associated with both chemotherapy resistance and invasion. In the current study we have utilized the C6 glioma cell line to investigate the response to long-term, clinically relevant application of topoisomerase I and II inhibitors. Treatment with etoposide resulted in an increase in resistance to this topoisomerase II inhibitor. By contrast, the continuous exposure to a topoisomerase I inhibitor did not result in increased drug resistance, but was associated with a reduction in cell migration. This data supports further investigation of topoisomerase I inhibition as a means to inhibit glioma invasion without the development of parallel chemoresistance

ABCB1 variants C3435T and T129C are not associated with colorectal cancer risk

Background: Colorectal cancer (CRC) is one of the most prevalent cancers in Saudi Arabia that is highly characterized with poor survival rate and advanced metastasis. Many studies contribute this poor outcome to the expression of ABC transporters on the surface of cancer cells. Objectives: In this study, two ABCB1 variants, C3435T and T129C, were examined to evaluate their contribution to CRC risk. Methods: 125 subjects (62 CRC patients and 63 healthy controls) were involved. The DNA was isolated and analyzed with PCR-RFLP to determine the different genotypes. The hardy-Weinberg equilibrium was performed to determine genotype distribution and allele frequencies. Fisher\u2019s exact test (two-tailed) was used to compare allele frequencies between patients and control subjects. Results: The study showed that for SNP C3435T, the population of both CRC patients and controls were out of Hardy-Weinberg equilibrium. Genotype distribution for CRC patients was (Goodness of fit \u3c72 = 20, df= 1, P 640.05), whereas, for the controls the genotype distribution was (Goodness of fit \u3c72 = 21, df =1, P 640.05). For SNP T129C, all subjects showed normal (TT) genotype. Conclusion: There was no significant association between ABCB1 3435C>T and 129T>C polymorphisms with CRC risk

Comparative Study of the Antioxidant Activity of Two Popular Green Tea Beverages Available in the Local Market of Saudi Arabia

Abstract Antioxidants have numerous applications due to their multiple roles in diminishing harmful effects of oxidative stress. The objective of this work was to highlight the importance of green tea by evaluating the antioxidant activity of the most popular green tea brands in Saudi Arabia, Lipton and Rabea. To our knowledge, no studies have so far been done to estimate the antioxidant activity of these brands. To determine the antioxidant activities of these two brands, 10 mg/ml of each brand was extracted and their total phenolic content (TPC), 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity, hydrogen peroxide scavenging activity, ferric reducing power and ferrous ion chelating effect were measured. The TPC of Lipton tea was 678.7 µg of gallic acid equivalents (GAE)/10mg, whereas in Rabea tea, the TPC was 647.1 µg GAE/10mg. The presence study indicated that there were no significant differences in total phenolic contents and the percentage inhibition as shown in DPPH and H 2 O 2 assays among Lipton and Rabea green teas. Moreover, it was found that all assays have exhibited high antioxidant activity in both green teas. In conclusion, our study showed evidence for evenness and stability of the antioxidant activity of the two commercial green teas available in the markets of Saudi Arabia. Continued researches are needed to further the current knowledge on the health-promoting effects of this popular beverage using different supplements by different mechanisms

Drug resistance mechanisms in a high grade glioma cell line

Glioblastoma multiforme (GBM) is the most common and aggressive primary brain tumour. Despite advances in GBM treatment, there is a high frequency of local relapse due to the acquisition of drug resistance. Investigation of glioma cell lines will help us to understand the molecular basis of this hard to treat tumour. In this study, the rat C6 glioma cell line was used as a model alongside two drug selected derivatives (C6-etoposide and C6-irinotecan) to investigate the mechanisms of chemo-resistance in glioma by identifying candidate proteins, genes, and key signalling pathways. Proteomic (2D gel electrophoresis) and genomic (gene array) analyses were performed to determine protein and gene expression changes. Integration of this data with cellular pathway analysis resulted in the prediction that cellular migration and the response to oxidative stress would be distinct in the drug selected C6 cell lines. Cell migration was subsequently assessed using wound scratch repair and transwell migration assays, whilst the response to oxidative stress produced by reactive oxygen species was determined fluorimetrically. The C6 cell line exposed to irinotecan (DNA topoisomerase I inhibitor) showed reduced migration, even under the influence of chemoattractant, compared to other cell lines, consistent with alterations in the expression of collagen genes. The C6 cell line exposed to etoposide (DNA topoisomerase II inhibitor) showed greater resistance to oxidative stress which was proposed to be due to alterations in the signalling pathways downstream of the PTEN/PI3Kinase. Future studies, investigating the effect of PI3Kinase pathway inhibitors are considered and it is proposed that further research into this signalling pathway will be able to uncover the molecular basis of distinct chemo-resistance in this important model cell system for aggressive glioma

Drug resistance mechanisms in a high grade glioma cell line

Glioblastoma multiforme (GBM) is the most common and aggressive primary brain tumour. Despite advances in GBM treatment, there is a high frequency of local relapse due to the acquisition of drug resistance. Investigation of glioma cell lines will help us to understand the molecular basis of this hard to treat tumour. In this study, the rat C6 glioma cell line was used as a model alongside two drug selected derivatives (C6-etoposide and C6-irinotecan) to investigate the mechanisms of chemo-resistance in glioma by identifying candidate proteins, genes, and key signalling pathways. Proteomic (2D gel electrophoresis) and genomic (gene array) analyses were performed to determine protein and gene expression changes. Integration of this data with cellular pathway analysis resulted in the prediction that cellular migration and the response to oxidative stress would be distinct in the drug selected C6 cell lines. Cell migration was subsequently assessed using wound scratch repair and transwell migration assays, whilst the response to oxidative stress produced by reactive oxygen species was determined fluorimetrically. The C6 cell line exposed to irinotecan (DNA topoisomerase I inhibitor) showed reduced migration, even under the influence of chemoattractant, compared to other cell lines, consistent with alterations in the expression of collagen genes. The C6 cell line exposed to etoposide (DNA topoisomerase II inhibitor) showed greater resistance to oxidative stress which was proposed to be due to alterations in the signalling pathways downstream of the PTEN/PI3Kinase. Future studies, investigating the effect of PI3Kinase pathway inhibitors are considered and it is proposed that further research into this signalling pathway will be able to uncover the molecular basis of distinct chemo-resistance in this important model cell system for aggressive glioma.EThOS - Electronic Theses Online ServiceGBUnited Kingdo

Drug resistance mechanisms in a high grade glioma cell line

Glioblastoma multiforme (GBM) is the most common and aggressive primary brain tumour. Despite advances in GBM treatment, there is a high frequency of local relapse due to the acquisition of drug resistance. Investigation of glioma cell lines will help us to understand the molecular basis of this hard to treat tumour. In this study, the rat C6 glioma cell line was used as a model alongside two drug selected derivatives (C6-etoposide and C6-irinotecan) to investigate the mechanisms of chemo-resistance in glioma by identifying candidate proteins, genes, and key signalling pathways. Proteomic (2D gel electrophoresis) and genomic (gene array) analyses were performed to determine protein and gene expression changes. Integration of this data with cellular pathway analysis resulted in the prediction that cellular migration and the response to oxidative stress would be distinct in the drug selected C6 cell lines. Cell migration was subsequently assessed using wound scratch repair and transwell migration assays, whilst the response to oxidative stress produced by reactive oxygen species was determined fluorimetrically. The C6 cell line exposed to irinotecan (DNA topoisomerase I inhibitor) showed reduced migration, even under the influence of chemoattractant, compared to other cell lines, consistent with alterations in the expression of collagen genes. The C6 cell line exposed to etoposide (DNA topoisomerase II inhibitor) showed greater resistance to oxidative stress which was proposed to be due to alterations in the signalling pathways downstream of the PTEN/PI3Kinase. Future studies, investigating the effect of PI3Kinase pathway inhibitors are considered and it is proposed that further research into this signalling pathway will be able to uncover the molecular basis of distinct chemo-resistance in this important model cell system for aggressive glioma.EThOS - Electronic Theses Online ServiceGBUnited Kingdo

Genetic Variants in the Mitochondrial Thymidylate Biosynthesis Pathway Increase Colorectal Cancer Risk

We assess the contributions of genetic variants for the enzymes involved in capecitabine metabolism to colorectal cancer (CRC) development risk. In this case-control study, DNA samples were collected from 66 patients (King Abdulaziz University Hospital) and 65 controls (King Fahad General Hospital) between April and November 2022 to be used in PCR-RFLP. The chi-square (χ2) test at a significance level of p ˂ 0.05 was used to estimate genotype and allele frequencies. The Lys27Gln variant of cytidine deaminase (CDA) showed a risk ratio (RR) of 1.47 for heterozygous (AC) carriers, with genotype distributions for patients (χ2 = 1.97) and controls (χ2 = 14.7). Homozygous (AA) Ala70Thr carriers demonstrated a three-fold higher risk, with genotype distributions for patients (χ2 = 3.85) and controls (χ2 = 4.23). Genotype distributions of the 5,10-methylenetetrahydrofolate reductase (MTHFR) C677T variant for patients were (χ2 = 22.43) and for controls were (χ2 = 0.07); for the MTHFR A1298C variant, they were (χ2 = 54.44) for patients and (χ2 = 4.58) for controls. Heterozygous (AC) carriers of the A1298C variant demonstrated highly significant protection against CRC development (RR = 0.2, p = 0.001), while a two-fold higher risk for CRC was estimated for homozygous genotype (CC) carriers. In conclusion, the heterozygous genotype of CDA Lys27Gln, the homozygous genotype of CDA Ala70Thr, and the homozygous genotype of MTHFR A1298C were associated with CRC development risk. The heterozygous genotype of MTHFR A1298C variant provided highly significant protection against CRC development. Further examinations using a larger population size are needed to reliably confirm our findings

ASSESSMENT OF THE ANTIOXIDANT PROPERTIES OF THE MOST COMMON COFFEE BREWS AVAILABLE IN THE LOCAL MARKETS OF THE WESTERN REGION OF SAUDI ARABIA Journal of Experimental Biology and Agricultural Sciences

KEYWORDS ABSTRACT The purpose of the current study is to determine the antioxidant activity of the most commonly used coffee brews in the Western region of Saudi Arabia. Further, total phenolic and flavonoid contents were measured and antioxidants properties including ferric reducing antioxidant power and ferrous ion chelating activity, DPPH radical scavenging activity, and scavenging of hydrogen peroxide assays were also determined. Results of study revealed that phenolic contents were 741, 835 and 578 µg/ml of gallic acid/2 g of coffee, and the total flavonoid contents were 711, 802, and 828 µg/ml of catechin/2 g of coffee for Nescafé red mug, Turkish coffee and Espresso, respectively. The inhibition percentage of hydrogen peroxide showed highly significant reduction (p<0.001) in Turkish coffee compared to Nescafé red mug. Further, DPPH activity also showed highly significant reduction (p<0.01) in Espresso and Turkish coffee compared to Nescafé red mug, whereas, Espresso showed a highly significant increase (p<0.01) in reducing power activity. Regarding the metal chelating activity, Espresso showed a significant decrease (p<0.05) in metal chelation activity as compared to Nescafé red mug. The current study demonstrated that Nescafé red mug, Turkish coffee and Espresso, were the most consumed coffee types in the Western region of Saudi Arabia according to the survey. These three types of coffee showed high phenolic and flavonoid contents as well as a high antioxidant activity

Addition of evaporated milk alters antioxdant properties of most commonly consumed coffee in Saudi Arabia

Coffee is one of the most commonly consumed beverages worldwide. The aim of current study is to evaluate the effect of adding evaporated milk (full fat and low fat) on the antioxidant properties of coffee and to assess the possible alterations of polyphenols bioavailability when evaporated milk is added. To determine the effects of adding milk on the antioxidant activities of instant coffee, total polyphenols and flavonoids contents, 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity, hydrogen peroxide scavenging activity, ferric reducing power and ferrous ion chelating effect were determined. Total polyphenol and flavonoid contents were significantly higher (p<0.0001) in both coffee milk samples, coffee with full fat evaporated milk (CFEM) and coffee with low fat evaporated milk (CLEM), in comparison to pure coffee. Moreover, DPPH scavenging activity was significantly increased (p<0.001) in CFEM and CLEM whereas, hydrogen peroxide radical scavenging activity and ferric reducing power were significantly decreased (p<0.001) in CFEM and CLEM samples. In conclusion, our study showed that adding different concentrations of evaporated milk to instant coffee influences the bioavailability and efficacy of coffee antioxidants. This might be due to the presence of proteins and fats in milk