The role of a putative tumor suppressor migration and invasion inhibitory protein in prostate cancer
Hopeasaari, Eevaliisa (2015)
2015
Biotekniikan koulutusohjelma
Luonnontieteiden tiedekunta - Faculty of Natural Sciences
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Hyväksymispäivämäärä
2015-06-03
Julkaisun pysyvä osoite on
https://urn.fi/URN:NBN:fi:tty-201505191307
https://urn.fi/URN:NBN:fi:tty-201505191307
Tiivistelmä
Prostate cancer is the most common cancer in Western countries. As the disease proceeds to castration resistant phase, it often leads to death due to lack of effective treatment methods and lack of knowledge of molecular mechanisms leading to it. Migration and invasion inhibitory protein (MIIP) has been shown to have tumor suppressing effects in other cancers. The effect of it in prostate cancer has not been previously studied. The aims of this study were to discover the effect of MIIP on growth and migration of prostate cancer cells and to detect possible mutations and a single nucleotide polymorphism (SNP) rs2295283 previously shown to affect breast cancer risk from the gene.
DU145 and LAPC-4 prostate cancer cell lines were used. DU145 cells were stably transfected with a plasmid containing MIIP and an empty control plasmid. LAPC-4 cells were transfected with small interfering RNAs (siRNAs) targeting MIIP and a control siRNA. Proliferation of the cells was studied with image analysis and alamarBlue methods. Migration was studied with scratch assay. Mutations and SNPs were studied by sequencing.
Clear results from the effects of silencing MIIP in LAPC-4 cells were not achieved. In the proliferation assays, image analysis and alamarBlue methods gave contradictory result and due to difficulties in image analysis, results from alamarBlue should be considered more reliable. AlamarBlue assay showed the metabolic activity of DU145 cells overexpressing MIIP to be significantly lower than in cells transfected with control vector. However, DU145 cells overexpressing MIIP migrated significantly faster that cells transfected with control vector. Mutations in the gene were not detected and SNP rs2295283 was not found to be associated with prostate cancer risk.
It seems unlikely that MIIP would have a significant effect in most prostate cancer cases. However, it is possible that the role of MIIP is more notable in some of prostate cancer subtypes. More experiments are needed to fully uncover the effect of MIIP in prostate cancer cells.
DU145 and LAPC-4 prostate cancer cell lines were used. DU145 cells were stably transfected with a plasmid containing MIIP and an empty control plasmid. LAPC-4 cells were transfected with small interfering RNAs (siRNAs) targeting MIIP and a control siRNA. Proliferation of the cells was studied with image analysis and alamarBlue methods. Migration was studied with scratch assay. Mutations and SNPs were studied by sequencing.
Clear results from the effects of silencing MIIP in LAPC-4 cells were not achieved. In the proliferation assays, image analysis and alamarBlue methods gave contradictory result and due to difficulties in image analysis, results from alamarBlue should be considered more reliable. AlamarBlue assay showed the metabolic activity of DU145 cells overexpressing MIIP to be significantly lower than in cells transfected with control vector. However, DU145 cells overexpressing MIIP migrated significantly faster that cells transfected with control vector. Mutations in the gene were not detected and SNP rs2295283 was not found to be associated with prostate cancer risk.
It seems unlikely that MIIP would have a significant effect in most prostate cancer cases. However, it is possible that the role of MIIP is more notable in some of prostate cancer subtypes. More experiments are needed to fully uncover the effect of MIIP in prostate cancer cells.