Alu and Poet Ankyrine Detection and Quantization in Cell Free Dna of Cancer Patients
Cancer Research Journal
Volume 6, Issue 1, March 2018, Pages: 20-25
Received: Dec. 21, 2017;
Accepted: Jan. 15, 2018;
Published: Mar. 5, 2018
Views 760 Downloads 54
Ahmed Darweesh Jabbar, Biology Department, College of Science, Wasit University, Wasit, Iraq
Safa Abdul Ilah Faraj, Faculty of medicine, Wasit University, Wasit, Iraq
Tariq Dawood Salman, Oncology and Haematology Center, AL-Karamah Teaching Hospital, Wasit, Iraq
Sarah Majed Kadhum, Biology Department, College of Science, Wasit University, Wasit, Iraq
Zainab Dhiyaa Jabuori, Biology Department, College of Science, Wasit University, Wasit, Iraq
In this study, specimens of plasma were collected from 96 from patient diagnosed with cancer from Al Karama teaching hospital. Also plasma collected from 25 healthy individual as control. Cell-free DNA (cfDNA) was extracted from 1ml plasma using high pure viral Nucleic Acid Kit as an alternative to high expensive cfDNA kit. cfDNA extracted according to the manufacturer’s instructions, but without the use of carrier RNA. This represent the first recorded method of using viral nucleic acid extraction kit for collection of cell free DNA. Results indicate that all cancer samples show significant increase of cfDNA in compare with control (p 0.01). and this dramatic increase in DNA concentration provide good indicator about health condition. DNA integrity have been checked depending on amplification of Arthrobacter luteus (Alu sequence). Both small (115) and large (247) large Alu stretches have amplified using RT PCR using SYBR green dye. Results shows that integrity of DNA extracted from patients and control are suitable for any further molecular investigation, and short Alu repeats are much less abundant in health individual in compare with cancer patient of all cancer types under study. Depending on these results one can conclude that almost all DNA obtained from normal patients is finely fragmented due to apoptosis action and that’s why give very little positive. results in compare with corresponding cancer patient which release large DNA fragments resulted from necrosis and NK/Tc cells activity, which is amplifiable more efficiently. In this work another molecular study conducted to investigate the cancer specific sequences. Since we deals with different cancers type in this study, two types of prostate, ovary, testis expressed protein (POTE Ankyrine) were tested 2α and 2β. Results shows that almost all healthy group were negative for specific POTE test while cancer patient samples are positive for POTE specific amplification. This results are promising since it could be developed to be a building block of cfDNA cancer tests.
Ahmed Darweesh Jabbar,
Safa Abdul Ilah Faraj,
Tariq Dawood Salman,
Sarah Majed Kadhum,
Zainab Dhiyaa Jabuori,
Alu and Poet Ankyrine Detection and Quantization in Cell Free Dna of Cancer Patients, Cancer Research Journal.
Vol. 6, No. 1,
2018, pp. 20-25.
Alix-Panabieres C, Pantel K. (2014). The circulating tumor cells: liquid biopsy of cancer. Klin Lab Diagn; (4):60-4.
Mandel, P. and Metais, P. (1948). Les acides nucleiques du plasma sanguin chez l’homme. C. R. Seances Soc. Biol. Fil., 142, 241–243.
Zhang J, Tong KL, Li PKT, Chan AYW, Yeung CK, Pang CCP, et al. (1999). Presence of Donor- and Recipient-derived DNA in Cell- free Urine Samples of Renal Transplantation Recipients: Urinary DNA Chimerism. Clin Chem; 45:1741-6.
Jiang WW, Masayesva B, Zahurak M, Carvalho AL, Rosenbaum E, Mambo E, et al. (2005). Increased Mitochondrial DNA Content in Saliva Associated with Head and Neck Cancer. Clin Cancer Res; 11:2486-91.
Diehl F, Schmidt K, Durkee KH, Moore KJ, Goodman SN, Shuber AP, et al. (2008). Analysis of Mutations in DNA Isolated From Plasma and Stool of Colorectal Cancer Patients. Gastroenterology; 135:489-98.
Leon SA, Revach M, Ehrlich GE, Adler R, Petersen V, Shapiro B. (1981). DNA in synovial fl uid and the circulation of patients with arthritis. Arthritis Rheum; 24:1142-50.
Rhodes CH, Honsinger C, Sorenson GD. (1994). Detection of tumorderived DNA in cerebrospinal fl uid. J Neuropathol Exp Neurol; 53:364-8.
Hickey KP, Boyle KP, Jepps HM, Andrew AC, Buxton EJ, Burns PA. (1999). Molecular detection of tumour DNA in serum and peritoneal fl uid from ovarian cancer patients. Br J Cancer; 80:803–8.
Anker P, Stroun M. (2000). Circulating DNA in plasma or serum. Medicina (B Aires); 60:699-702.
Jahr S, Hentze H, Englisch S, Hardt D, Fackelmayer FO, Hesch RD, Knippers R. (2001). DNA Fragments in the Blood Plasma of Cancer Patients: Quantitations and Evidence for Their Origin from Apoptotic and Necrotic Cells. Cancer Res; 61 (4):1659–65.
Tokuhisa, Y., Iizuka, N., Sakaida, I., Moribe, T., Fujita, N., Miura, T. et al. (2007). Circulating cell-free DNA as a predictive marker for dis-tant metastasis of hepatitis C virus-related hepatocellular carcinoma. British Journal of Cancer, 97, 1399-1403.
Bettegowda C, Sausen M, Leary RJ, Kinde I, Wang Y, Agrawal N, Bartlett BR, Wang H, Luber B, Alani RM, Antonarakis ES, Azad NS, Bardelli A, et al. (2014). Detection of circulating tumor DNA in early- and late-stage human malignancies. Sci Transl Med.; 6: 2 24-24.
Roth C, Pantel K, Müller V, Rack B, Kasimir-Bauer S, Janni W, Schwarzenbach H. (2011). Apoptosis-related deregulation of proteolytic activities and high serum levels of circulating nucleosomes and DNA in blood correlate with breast cancer progression. BMC Cancer.; 11: 4.
Lander ES, Linton LM, Birren B, Nusbaum C, Zody MC, Baldwin J, Devon K, Dewar K, Doyle M, FitzHugh W, Funke R, Gage D, Harris K, Heaford A, Howland J, Kann L, Lehoczky J, LeVine R, McEwan P, McKernan K, Meldrim J, Mesirov JP, Miranda C, Morris W, Naylor J, Raymond C, Rosetti M, Santos R, Sheridan A, Sougnez C, et al. (2001). Initial sequencing and analysis of the human genome. International Human Genome Sequencing Consortium. Nature, 409:860-921.
Dagan T, Sorek R, Sharon E, Ast G, Graur D (2004). AluGene: a database of Alu elements incorporated within protein-coding genes. Nucleic Acids Res 32: D489–D492.
Bera, T. K., et al.,(2002). POTE, a highly homologous gene family located on numerous chromosomes and expressed in prostate, ovary, testis, placenta, and prostate cancer. Proc. Natl. Acad. Sci. U. S. A. 99, 16975–16980.
Naoyuki Umetani, Joseph Kim, Suzanne Hiramatsu, HowardA. Reber, Oscar J. Hines, Anton J. Bilchik, Dave S. B. Hoon (2016). Increased Integrity of Free Circulating DNA in Sera of Patients with Colorectal or Periampullary Cancer: Direct Quantitative PCR for ALU Repeats. Clinical Chemistry 52:6 1062–1069.
Schwarzenbach, H.; Hoon, D. S. B.; Pantel, K. (2011). Cell-free nucleic acids as biomarkers in cancer patients. Nat. Rev. Cancer, 11, 426–437.
Wang BG, Huang HY, Chen YC, et al. (2003). Increased plasma DNA integrity in cancer patients. Cancer Res 63:3966-3968.