Uğur GÖNÜLLÜ, Elif ŞEN, Nejat AKAR

The detection of quantitative serum p53 protein in lung cancer

Tüm insan tümörlerinin yarısından fazlasında hücre içinde biriktiği gösterilen p53 proteininin, çeşitli maligniteleri olan hastaların serumlarında değişken olarak bulunduğu bildirilmiştir. Bu çalışmada, akciğer kanserli hastaların serumlarında p53 proteininin saptanması ve akciğer kanserinde p53 değişimlerinin belirleyicisi olarak değerinin doğrulanması amaçlanmıştır. Yeni tanı almış 94 akciğer kanserli hastada serum p53 proteinin saptanmasında pantropik kantitatif ELISA tekniği kullanıldı. Serum örnekleri herhangi bir tedavi almadan önce, başvuru sırasında alındı. Otuzdört sağlıklı gönüllüden oluşan kontrol grubunda; serum p53 proteini saptanmadı. Serum p53 proteini; 94 hastanın sadece 3 (%3.2)’ünün serumlarında vardı. Küçük hücreli dışı akciğer kanserli grupta 72 hastanın 2 (%2.8)’sinde ve küçük hücreli akciğer kanseri olan 22 hastanın 1 (%4.5)’inde serum p53 proteini saptandı. Serum p53 proteini düzeyleri pozitif olan örneklerde 1-31.25 U/mL arasında değişiyordu. Serumlarında p53 proteini saptananlar ileri evre hastalığı olan ve kötü prognozlu hastalardı. Sonuç olarak; serum p53 protein düzeyinin prognostik önemi p53 proteini pozitif olan hastaların sayısının az olması nedeniyle belirlenemedi. Kantitatif olarak ELISA ile serum p53 proteini analizinin kullanımı akciğer kanserli hastalarda kolay uygulanabilir bir teknik olmasına karşın, p53 proteini değişikliklerini değerlendirmede bir belirleyici olarak sınırlı bir kullanıma sahiptir.

Akciğer kanserinde kantitatif serum p53 proteini saptanması

p53 protein, which accumulates intracellularly in over half of all human tumors, has been reported to be variably present in the sera of patients with various malignancies. In this study, it was aimed to detect p53 protein in the sera of lung cancer patients, and to verify its value as a marker of p53 alterations in lung cancer. A pantropic quantitative ELISA technique was used to detect serum p53 protein of 94 newly diagnosed patients with lung cancer. Serum samples were collected on admission before any treatment. There was no detectable serum p53 protein in the control group including 34 healthy volunteers. Serum p53 protein was present in only 3 (3.2%) of 94 patients. In nonsmall cell lung cancer (NSCLC) group, serum p53 protein had been detected in 2 (2.8%) of 72 patients, and it was detectable in 1 (4.5%) of 22 patients in SCLC group. Serum levels of p53 protein ranged from 1 U/mL to 31.25 U/mL in positive samples. Patients who had p53 protein in their serum samples, were at late stage and had poor prognosis. In conclusion; prognostic value of detectable serum p53protein levels could not be define, because of the small number of p53 positive patients. The use of quantitative serum p53 protein analysis with ELISA is of very limited value as a marker in evaluating p53 changes in lung cancer patients, despite the fact that is an easy technique to perform.

PDF

___

1.Peto R, Lopez AD, Boreham J, et al. Mortality from smoking worldwide. Br Med Bull 1996; 52: 12-21.
2.Jemal A, Thomas A, Murray T, Thun M. Cancer statistics,2002. CA Cancer J Clin 2002; 52:23-47.
3.Parkin DM, Pisanl P, Ferlay J. Estimates of the worldwide incidence of 25 major cancers in 1990. Int J Cancer 1999; 80: 827-41.
4.Brambilla E, Brambilla C. P53 and lung cancer. Path Biol 1997; 45: 852-63.
5.Greenblatt MS, Bennett WP, Hollstein M. Mutations in the p53 tumor suppressor gene: clues to cancer etiology and molecular pathogenesis. Cancer Res 1994; 54: 4855-78.
6.Hollstein M, Rice K, Greenblatt MS, et al. Database of p53 gene somatic mutations in human tumors and cell lines. Nucleic Acids Res 1994; 22: 3551-5.
7.Bennett WP, Hussain SP, Vahakangas K, et al. Molecular epidemiology of human cancer risk: gene-environment interactions and p53 mutation spectrum in human lung cancer. J Pathol 1999; 187: 8-18.
8.Tammemagi MC, McLaughlin JR, Bull SB, et al. Metaanalyses of p53 tumor suppressor gene alterations and clinicopathological features in resected lung cancer. Cancer Epidemiol Biomarkers Prev 1999; 8: 625-34.
9.Soussi T. The p53 tumor suppressor gene: from molecular biology to clinical investigation. In: Lenfant C (ed).Lung Tumors. New York: Marcel-Dekker, 1999:453-72.
10.Dowell SP, Wilson PO, Derias NW, et al. Clinical utility of the immunocytochemical detection of p53 protein in cytological specimens. Cancer Res 1994; 54:2914-8.
11.Hall PA, Lane DP. p53 in tumour pathology-can we trust immunohistochemistry-revisited. J Pathol 1994; 172:1-4.
12.Midgley CA, Fisher CJ, Bartek J, et al. Analysis of p53 expression in human tumors: an antibody raised against human p53 expression in Escherichia coli. J Cell Sci 1992; 101: 183-9.
13.Vojtesek B, Bartek J, Midgley CA, Lane DP. An immunochemical analysis of human nuclear phosphoprotein p53: new monoclonal antibodies and epitope mapping using recombinant p53. J Immunol Methods 1992; 151:237-44.
14.Hassapoglidou S, Diamandis EP, Sutherland DJ. Quantification of p53 protein in tumor cell lines, breast tissue extracts and serum with, time-resolued Immunofluorometry. Oncogene 1993; 8: 1501-9.
15.Soussi T. p53 antibodies in the sera of patienis with various types of cancer: a review. Cancer Res 2000;60:1777- 88.
16.Luo JC, Neugut Al, Garbowski G, et al. Levels of p53 antigen in the plasma of patients with adenomas and carcinomas of the colon. Cancer Lett 1995; 91:235-40.
17.Fontanini G, Bigini D, Vignati S, et al. Levels of p53 antigen in the serum of non-small cell lung cancer patients correlate with positive p53 immunohistochemistry on tumor sections, tumor necrosis and nodal involvement. Int J Oncol 1994; 5: 553-8.
18.Luo JC, Zehab R, Anttlla S, et al. Detection of serum p53 protein in lung cancer patients. J Occup Med 1994; 36: 155-60.
19.Levesque MA, D'Costa M, Diamandis EP, et al. P53 protein is absent from the serum of patients with lung cancer.Br J Cancer 1996; 74: 1434-40.
20.Segawa Y, Takigawa M, Mandai K, et al. Measurement of serum p53 protein in patients with small cell lung cancer and results of its clinicopathological evaluation.Lung Cancer 1997; 16: 229-38.
21.Morita T, Tachikawa M, Kumamaru T, et al. Serum anti-p53 antibodies and p53 protein status in the sera and tumors from bladder cancer patients. Eur Urol 2000; 37: 79-84.
22.Suwa H, Ohshio G, Okada M, et al. Clinical significance of serum p53 antigen in patients with pancreatic carcinomas. Gut 1997; 40: 647-53.
23.Barbati A, Mariani L, Porpora MG, et al. Serum evaluation of p53 protein in patients with gynaecological cancer.Anticancer Res 2000; 20: 1033-6.
24.Lehtinen T, Aine R, Kellokumpu-Lehtinen P, et al. Evaluation of plasma levels of thymidine kinase and mutated p53 in 81 patients with newly diagnosed malignant lymphoma. Ada Oncol 1993; 32: 779-81.
25.Trumper L, Jung W, Dahi C, et al. lnterleukin-7, interleukin-8, soluble TNF receptor, and p53 protein levels are elevated in the serum of patients with Hodgkin's disease. Ann Oncol 1994; 5 (Suppl 1): 93-6.
26.Rosanelli GP, Wirnsberger GH, Purstner P, Steindorfer P. DMA flow cytometry and immunohistochemical demonstration of mutant p53 protein versus TPS and mutant p53 protein serum levels in human breast cancer. Proc Am Assoc Cancer Res 1993; 34:227.
27. Mountain CF. Revisions in the International System for Staging Lung Cancer.Chest 1997;111: 1710-7.
28.Zelen M. Keynote address on bioistatistics and data retrieval.Cancer Chemother Rep 1973; 4: 31-42.
29.Levine AJ. p53, the cellular gatekeeper for growth and division. Cell 1997; 88: 323-31.
30.Shopland DR, Eyre HJ, Pechacek TF. Smoking-attributable cancer mortality in 1991: is lung cancer now the leading cause of death among smokers in United States.J Natl Cancer Inst 1991; 83: 1142-8.
31.Fisher CJ, Gillett CE, Vojtesek B, et al. Problems with immunohistochemical staining: the effect of fixatioft and variation in method of evaluation. Br J Cancer 1994; 69:26-31.
32.Crawford LV, Pim DC, Bullbrook RD. Detection of antibodies against the cellular protein p53 in sera from patients with breast cancer. Int J Cancer 1982; 30: 403-8.
33.Winter SF, Minna JD, Johnson BE, et al. Development of antibodies against p53 in lung cancer patients appears to be dependent on the type of p53 mutation. Cancer Res 1992; 52: 4168-74.
34.Kaur J, Srivastava A, Ralhan R. Serum p53 antibodies in patients with oral lesions: correlation with p53/HSP70 complexes. Int J Cancer 1997; 74:609-13.
35.Davidoff AM, Iglehart JD, Marks JR. Immune response to p53 is dependent upon p53/HSP70 complexes in breast cancers. Proc Nat Acad Sci 1992;89:3439-42