Aim of the research:To investigate whether the polymorphisms of GSTP1, GSTT1 and GSTM1 genes are associated with colorectal cancer in a sample of Polish patients with and without type 2 diabetes. Material and methods: Over 200 patients aged > 40 years were recruited and divided into 4 age-matched groups: patients with diagnosed type 2 diabetes, patients with diagnosed type 2 diabetes and colorectal cancer, patients with colorectal cancer and without type 2 diabetes and patients without type 2 diabetes and without colorectal cancer. DNA for genetic testing was isolated from blood samples. The GST gene polymorphism was examined using qPCR method, with TaqMan probes. It included GSTP1 SNP Ile105Val (rs1695) polymorphism, and deletion of copies of GSTT1 and GSTM1 genes (genotypes GSTT1 null/null and GSTM1 null/null). Results: The analysis of the frequency of GST gene polymorphisms in the group of patients with type 2 diabetes and colorectal cancer compared to patients with type 2 diabetes showed no statistically significant differences. There were also no statistically significant differences in the distribution of GST gene polymorphisms between patients with diabetes or/and colorectal cancer and population without these diseases. Conclusions: Extending the analysis to further genetic and environmental factors and taking into account their mutual interactions is needed to search for the relationship between type 2 diabetes and an increased risk of colorectal cancer.
StreszczenieCel pracy: Określenie, czy polimorfizmy genów GSTP1, GSTT1 i GSTM1 są związane z rakiem jelita grubego w populacji polskich pacjentów z cukrzycą typu 2 i bez cukrzycy. Materiał i metody: Ponad 200 pacjentów powyżej 40. roku życia zostało zrekrutowanych do badania i podzielonych na 4 grupy dopasowane wiekowo: pacjenci ze zdiagnozowaną cukrzycą typu 2, pacjenci ze zdiagnozowanymi cukrzycą typu 2 i rakiem jelita grubego, pacjenci z rakiem jelita grubego, bez cukrzycy typu 2, pacjenci bez raka jelita grubego i bez cukrzycy. DNA do badań genetycznych izolowano z próbek krwi. Polimorfizm genów GST badano z użyciem metody qPCR i sond typu Taqman. Badanie obejmowało polimorfizm typu SNP Ile105Val (rs 1695) oraz delecje kopii genów GSTT1 i GSTM1 (genotypy GSTT1 null/null oraz GSTM1 null/null). Wyniki: Analiza częstości występowania polimorfizmów genów GST u pacjentów z cukrzycą typu 2 i rakiem jelita grubego w porównaniu z pacjentami z cukrzycą typu 2 bez raka nie wykazała statystycznie istotnych różnic. Nie stwierdzono również istotnych różnic w dystrybucji polimorfizmów genów GST pomiędzy osobami z cukrzycą i/lub chorymi na raka jelita grubego a grupą bez tych schorzeń. Wnioski: W celu określenia związku pomiędzy cukrzycą typu 2 a zwiększonym ryzykiem wystąpienia raka jelita grubego konieczne jest rozszerzenie analizy o kolejne czynniki genetyczne i środowiskowe z uwzględnieniem ich wzajemnych interakcji.
Some of the products for the molecular diagnosis of infections do not have an endogenous internal control, and this is necessary to ensure that the result is not a false negative. The aim of the project was to design a simple low-cost RT-qPCR test that can confirm the expression of basic metabolism proteins, thus confirming the quality of genetic material for molecular diagnostic tests. Two successful equivalent qPCR assays for the detection of the GADPH and ACTB genes were obtained. The course of standard curves is logarithmic, with a very high correlation coefficient R2 within the range of 0.9955–0.9956. The reaction yield was between 85.5 and 109.7%, and the detection limit (LOD) with 95% positive probability was estimated at 0.0057 ng/µL for GAPDH and 0.0036 ng/µL for ACTB. These tests are universal because they function on various types of samples (swabs, cytology, etc.) and can complement the diagnosis of SARS-CoV-2 and other pathogens, as well as potentially oncological diagnostics.
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