DETECTION OF OXIDATIVE STRESS, APOPTOSIS AND MOLECULAR LESIONS IN HUMAN OVARIAN CANCER CELLS
Background. Ovarian cancer has the highest mortality rate of gynaecological cancers. This is partly due to the lack of effective screening markers. Indices of oxidative stress are well-recognized prognostic criteria for tumorous transformation of tissue, but their value depends on the type of tumor and the stage of its development.
Objective. The aim of this study is to clarify the relationship between antioxidant/pro-oxidant ratio and the signs of molecular lesions and apoptosis rate in blood of ovarian cancer patients and non-cancer ones.
Results. The ovarian cancer group is marked by antioxidant/prooxidant balance shifting to oxidative damage in blood as the consequence of overexpression of oxyradicals (by 300%). Higher level of glutathione (by 366%), lower level of metallothioneins (by 65%) as well as higher level of lipid peroxidation (by 174%) and protein carbonyls (by 186%) in blood of ovarian cancer patients compared to the normal ovarian group have been observed. The signs of cytotoxicity are determined in blood of ovarian cancer patients: an increased (compared to control) level of DNA fragmentation (by 160%), choline esterase (up to twice), higher rate of both caspase dependent and caspase independent lysosomal mediated apoptosis.
Conclusions. Cathepsin D activity both total and free, choline esterase activity, TBA-reactive substance and protein carbonyls level in blood could be used as the predictive markers of worse prognosis and the signs of human ovarian cancer.
Howlader N, Noone AM, Krapcho M, Garshell J, Neyman N, Altekruse SF et al. SEER Cancer Statistics Review, 1975-2010, National Cancer Institute. Bethesda, MD, http://seer.cancer.gov/csr/1975_2010/, based on November 2012 SEER data submission, posted to the SEER web site (accessed, April 2013). Controlled Trial. JAMA 2011; 305: 2295–2303.
Klaunig JE, Kamendulis LM, Hocevar BA. Oxidative stress and oxidative damage in carcinogenesis. Toxicol Pathol 2010; 38: 96–109.
Mahalingaiah PKS, Singh KP. Chronic oxidative stress increases growth and tumorigenic potential of MCF-7 breast cancer cells. PLOS ONE 2014; 9: e93799.
Reuter S, Gupta SC, Chaturvedi MM, Aggarwal BB. Oxidative stress, inflammation, and cancer: How are they linked? Free Radic Biol Med 2010; 49: 1603–1616.
Valko M, Rhodes CJ, Moncol J, Izakovic M, Mazur M. Free radicals, metals and antioxidants in oxidative stress-induced cancer. Chem Biol Interact 2006; 160: 1–40.
Visconti R, Grieco D. New insights on oxidative stress in cancer. Curr Opin Drug Discov Devel 2009; 12: 240–245.
Falfushynska H, Gnatyshyna L, Shulgai A, Shidlovsky V, Stoliar O. Oxidative stress in human thyroid gland in cases of iodine deficiency nodular goitre: from harmlessness to hazard depending on copper and iodine subcellular distribution. Int J Med Medical Res 2015; 1: 5–11.
Falfushynska HI, Gnatyshyna LL, Osadchuk OY, Shidlovsky VO, Stoliar OB. Trace elements storage and metallothioneins function in cases if human thyroid gland transformation. Ukr Biochem J 2014; 86: 107–113.
Falfushynska HI, Gnatyshyna LL, Deneha HV, Osadchuk OY, Stoliar OB. Manifestations of oxidative stress and molecular damages in ovarian cancer tissue. Ukr Biochem J 2015; 87: 93–102.
Beauchamp C, Fridovich I. Superoxide dismutase: improved assay and an assay applicable to acrylamide gels. Anal Biochem 1971; 44: 276–287.
Anderson ME. Determination of glutathione and glutathione disulfide in biological samples. Meth Enzymol 1985; 113: 548–555.
Lushchak VI, Bagnyukova TV, Lushchak OV. Indices of oxidative stress. 1. TBA-reactive substances and carbonylproteins. Ukr Biochem J 2004; 76: 136–141.
Ohkawa H, Ohishi N, Tagi K. Assay for lipid peroxides in animal tissues by thiobarbituric acid reaction. Anal Biochem 1979; 95: 351–358.
Viarengo A, Burlando B, Cavaletto M, Marchi B, Ponzano E, Blasco J. Role of metallothionein against oxidative stress in the mussel Mytilus galloprovincialis. Am J Physiol 1999; 277: 1612–1619.
Viarengo A, Ponzano E, Dondero F, Fabbri R. A simple spectrophotometric method for metallothionein evaluation in marine organisms: an application to Mediterranean and Antarctic molluscs.
Mar Environ Res 1997; 44: 69–84.
Ellman GL, Courtney KD, Andres VJ, Featherstone RM. A new and rapid colorimetric determination of acetylcholinesterase activity. Biochem Pharmacol 1961; 7: 88–95.
Olive PL. DNA precipitation assay: a rapid and simple method for detecting DNA damage in mammalian cells. Environ Mol Mutagen 1988; 11: 487–495.
Bonomini M, Dottori S, Amoroso A, Arduini A, Sirolli V. Increased platelet phosphatidylserine exposure and caspase activation in chronic uremia. J Thromb Haemost 2004; 2: 1275–1281.
Falfushynska H, Gnatyshyna L, Fedoruk O, Mitina N, Zaichenko A, Stoliar O, Stoika R. Hepatic metallothioneins in molecular responses to cobalt, zinc, and their nanoscale polymeric composites in frog Rana ridibunda. Comp Biochem Physiol 2015; 172-173: 45–56.
Dingle JT, Barrett AJ, Weston PD. Cathepsin D. Characteristics of immunoinhibition and the confirmation of a role in cartilage breakdown. Biochem J 1971; 123: 1–13.
Yu C, Huang X, Xu Y, Li H, Su J, Zhong J et al. Lysosome dysfunction enhances oxidative stressinduced apoptosis through ubiquitinated protein accumulation in Hela cells. Anat Rec (Hoboken) 2013; 296: 31–39.
Stoka V, Turk B, Schendel SL, Kim TH, Cirman T, Snipas SJ et al. Lysosomal protease pathways to apoptosis. J Biol Chem 2001; 276: 3149–3157.
Herr I, Debatin K-M. Cellular stress response and apoptosis in cancer therapy. Blood 2001; 98: 9.
Faraglia B, Bonsignore A, Scaldaferri F, Boninsegna A, Cittadini A, Mancuso C, Sgambato A. Caspase-3 inhibits the growth of breast cancer cells independent of protease activity. J Cell Physiol 2005; 202: 478–482.
Devarajan E, Sahin AA, Chen JS, Krishnamurthy RR, Aggarwal N, Brun AM et al. Down-regulation of caspase 3 in breast cancer: a possible mechanism for chemoresistance. Oncogene 2002; 21: 8843–8851.
Ruiz-Espejo F, Cabezas-Herrera J, Illana J, Campoy FJ, Vidal CJ. Cholinesterase activity and acetylcholinesterase glycosylation are altered inhuman breast cancer. Breast Cancer Res Treat 2002; 72: 11–22.
Zakut H, Ehrlich G, Ayalon A, Prody CA, Malinger G, Seidman S et al. Acetylcholinesterase and butyrylcholinesterase genes coamplify in primary
ovarian carcinomas. J Clin Invest 1990; 86: 900–908.
Fuhrman B, Partoush A, Aviram M. Acetylcholine esterase protects LDL against oxidation. Biochem Biophys Res Commun 2004; 322:974–978.
Bay B-H, Jin R, Huang J, Tan P-H. Metallothionein as a prognostic biomarker in breast cancer. Exp Biol Med 2006; 231: 1516–1521.
Takeda A, Hisada H, Okada S, Mata JE, Ebadi M, Iversen PL. Tumor cell growth is inhibited by suppressing metallothionein-I synthesis. Cancer Lett 1997; 116: 145–149.
Tekur S, Ho S-M. Ribozyme-mediated downregulation of human metallothionein II(a) induces apoptosis in human prostate and ovarian cancer cell lines. Mol Carcinog 2002; 33: 44–55.
Aiuchi T, Mihara S, Nakaya M, Masuda Y, Nakajo S, Nakaya K. Zinc ions prevent processing of caspase-3 during apoptosis induced by geranylgeraniol in HL-60 cells. J Biochem 1998; 124: 300–303.
Chimienti F, Seve M, Richard S, Mathieu J, Favier A. Role of cellular zinc in programmed cell death: temporal relationship between zinc depletion, activation of caspases, and cleavage of Sp family transcription factors. Biochem Pharmacol 2001; 62: 51–62.
Authors who sent their manuscript to International Journal of Medicine and Medical Research agree to the following terms:
1. Authors retain copyright and grant the journal right of first publication with the work simultaneously licensed under a Creative Commons Attribution License CC-BY-NC that allows others to share the work with an acknowledgment of the work's authorship and initial publication in this journal.
2. Authors able to enter into separate, additional contractual arrangements for the non-exclusive distribution of the journal's published version of the work (e.g., post it to an institutional repository or publish it in a book), with an acknowledgment of its initial publication in this journal.
3. Authors are permitted and encouraged to post their work online (e.g., in institutional repositories or on their website) prior to and during the submission process, as it can lead to productive exchanges, as well as earlier and greater citation of published work (See The Effect of Open Access).