Extra Virgin Olive Oil Impairs UV-Induced Cell Damage in Cancer-Prone Mice
INTRODUCTION
Ultraviolet radiation produces various wavelengths that are part of the electromagnetic spectrum. The four different UV wavelengths are classified as UVA, UVB, and UVC. UVA produces the longest wavelength at 320-400nm, UVB produces wavelengths from 290-320nm, and UVC wavelengths are the shortest wavelength that is completely absorbed by the ozone layer. However, it is known that UVA and UVB play an important role in premature skin aging, eye damage and most importantly skin cancers as the UV rays pierce the atmosphere. Studies have shown that UVB radiation is extremely mutagenic in animals compared to UVA radiation. …show more content…
The mouse tissue was collected 3 days after UVB exposure to allow the DNA lesions to be fixed into the DNA and expressed as stable mutations. Then high molecular weight DNA is extracted from tissues and purified. The shuttle vector which contains a target gene was recovered from genomic DNA with in vitro packaging extracts and the antioxidant (olive oil) was applied. (The mutations in the target gene are detected by a change in lambda phage plaque color.) They were plated for plaque formation at two temperatures. The mutant phage was stored at 24o C and 37oC. They were stored at two temperatures because the cll mutant phage will only form plaques at the first temperature while all phages will form plaques at the second mentioned temperature. The mutation frequency was then calculated by dividing the number of mutant plaques by the total number of plaques. The transgenic rodent mutation assay allows the lambda shuttle vector to be recovered and has the ability to measure the mutations in the lambda cll gene which will identify the rare mutations in mouse tissue from being exposed to UVB radiation as well as topical application of extra virgin olive …show more content…
Fetal bovine serum was used because it is a cell growth supplement which has a cell growth promoter for any cell and high levels of nutrients and a combination of growth factors. The cells were washed with PBS to ensure that the old medium/serum was removed in order to rule out protective effects due to the serum protein presence. It is useful to look at foci formation because the cells are plated at a high density and non-cancerous cells will not continue to grow when they reach neighboring cells. However, cancer cells grow and divide in an uncontrolled manner, so those cells will continue to keep growing even if there are cells next to them. The focus assay ultimately takes advantage of loss of contact inhibition. Studies that are in vivo are performed on a living organism, such as mice, and tumor growth can be recorded. The difference between in vivo and focus formation is that in vivo cells are studied inside the organism while the focus assay is when the cells are studied outside the
INTRODUCTION
Ultraviolet radiation produces various wavelengths that are part of the electromagnetic spectrum. The four different UV wavelengths are classified as UVA, UVB, and UVC. UVA produces the longest wavelength at 320-400nm, UVB produces wavelengths from 290-320nm, and UVC wavelengths are the shortest wavelength that is completely absorbed by the ozone layer. However, it is known that UVA and UVB play an important role in premature skin aging, eye damage and most importantly skin cancers as the UV rays pierce the atmosphere. Studies have shown that UVB radiation is extremely mutagenic in animals compared to UVA radiation. …show more content…
The mouse tissue was collected 3 days after UVB exposure to allow the DNA lesions to be fixed into the DNA and expressed as stable mutations. Then high molecular weight DNA is extracted from tissues and purified. The shuttle vector which contains a target gene was recovered from genomic DNA with in vitro packaging extracts and the antioxidant (olive oil) was applied. (The mutations in the target gene are detected by a change in lambda phage plaque color.) They were plated for plaque formation at two temperatures. The mutant phage was stored at 24o C and 37oC. They were stored at two temperatures because the cll mutant phage will only form plaques at the first temperature while all phages will form plaques at the second mentioned temperature. The mutation frequency was then calculated by dividing the number of mutant plaques by the total number of plaques. The transgenic rodent mutation assay allows the lambda shuttle vector to be recovered and has the ability to measure the mutations in the lambda cll gene which will identify the rare mutations in mouse tissue from being exposed to UVB radiation as well as topical application of extra virgin olive …show more content…
Fetal bovine serum was used because it is a cell growth supplement which has a cell growth promoter for any cell and high levels of nutrients and a combination of growth factors. The cells were washed with PBS to ensure that the old medium/serum was removed in order to rule out protective effects due to the serum protein presence. It is useful to look at foci formation because the cells are plated at a high density and non-cancerous cells will not continue to grow when they reach neighboring cells. However, cancer cells grow and divide in an uncontrolled manner, so those cells will continue to keep growing even if there are cells next to them. The focus assay ultimately takes advantage of loss of contact inhibition. Studies that are in vivo are performed on a living organism, such as mice, and tumor growth can be recorded. The difference between in vivo and focus formation is that in vivo cells are studied inside the organism while the focus assay is when the cells are studied outside the