Role of oxidative stress and regulation of D.N.A. methyl transferases during the acquisition of doxorubicin resistance in caki-1 kidney cancer cells

dc.contributor.committeeChairSingh, Kamaleshwar P.
dc.contributor.committeeMemberMayer, Greg
dc.contributor.committeeMemberAnderson, Todd
dc.creatorOludare, Charles Oladipo
dc.creator.orcid0000-0002-0211-6563
dc.date.accessioned2021-10-07T15:55:53Z
dc.date.available2021-10-07T15:55:53Z
dc.date.created2021-08
dc.date.issued2021-08
dc.date.submittedAugust 2021
dc.date.updated2021-10-07T15:55:55Z
dc.description.abstractThe main objective this paper is to understand the mechanistic basis of doxorubicin resistance in caki-1 kidney cancer cells from the oxidative stress and DNA methylation perspective. We exposed parental caki-1 cells to a repeated clinically relevant dose of doxorubicin over a period of 3 months and confirmed acquisition of doxorubicin resistance via MTT cell proliferation assay and flow cytometry. After confirming the development of a significant level of resistance to doxorubicin in the doxorubicin-exposed cells, we proceeded to evaluate the baseline level of intracellular oxidative stress in both the parental and resistant caki-1 cells and found that the resistant cells had a significantly lesser level of oxidative stress than the parental counterparts. We then tried to understand the role of antioxidant genes in observed lower level of oxidative stress and discovered that various antioxidant genes were significantly upregulated in the resistant cells. Meaning the reduction is oxidative stress was due to an increase in the cell’s antioxidant capacities We then tried to understand the role of transcription control in the upregulation of antioxidant genes. We check the level of both an antioxidant gene expression regulator/transcription factor (Nrf2) and a prooxidant gene transcription factor mtTFA and found that Nrf2 was much significantly upregulated while mtTFA was only mildly upregulated. This means that the upregulated antioxidant genes were due to an activation of transcriptional control. We also wanted to examine the role of DNA methylation genes in the acquisition of doxorubicin resistance and the role of DNA demethylation in restoring sensitivity to resistant cells. We proceeded to evaluate the expression of DNA methylation genes such as DNMT3a, DNMT3b and DNMT1 alongside MBD4 and found that all but DNMT3b were significantly upregulated in the resistant cells. After the confirming this we wanted to know if a pretreatment with a DNMT inhibitor 5-aza-2’-deoxycytidine will restore sensitivity to the resistant cells and found that the DNMT inhibitor significantly restored sensitivity to the resistant cells.
dc.description.abstractEmbargo status: Restricted until September 2026. To request an access exception, click on the PDF link to the left.
dc.format.mimetypeapplication/pdf
dc.identifier.urihttps://hdl.handle.net/2346/88054
dc.language.isoeng
dc.rights.availabilityRestricted until September 2026.
dc.subjectOxidative Stress
dc.subjectEpigenetics
dc.subjectDoxorubicin
dc.subjectResistance
dc.subjectAntioxidant
dc.titleRole of oxidative stress and regulation of D.N.A. methyl transferases during the acquisition of doxorubicin resistance in caki-1 kidney cancer cells
dc.typeThesis
dc.type.materialtext
local.embargo.lift2026-08-01
local.embargo.terms2026-08-01
thesis.degree.departmentEnvironmental Toxicology
thesis.degree.disciplineEnvironmental Toxicology
thesis.degree.grantorTexas Tech University
thesis.degree.levelMasters
thesis.degree.nameMaster of Science

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