Harzem ÖZGER, Levent ERALP, ATA CAN ATALAR, Berkin TOKER, Esberk Lora ATEŞ, Mustafa SUNGUR, Bilge BİLGİÇ, İnci AYAN

Osteosarkomlu hastalarda dirençle ilişkili proteinlerin prognoz ve sağkalım üzerine etkisi: İmmünhistokimyasal analiz

Amaç: Kemoterapi rejimlerindeki gelişmelere rağmen osteosarkom sağkalımında çok az ilerleme olmuştur. Bu çalışmada, osteosarkomlu hastalarda prognoz ile ilişkili bazı proteinlerin sağkalım ile ilişkisi değerlendirildi. Çalışma planı: Tedavisi ve izlemi hastanemizde yapılan 45 hastanın (24 erkek, 21 kadın) verileri geriye dönük olarak incelendi. Osteosarkom nedeniyle, neoadjuvan kemoterapi sonrasında, 41 hastaya ekstremite koruyucu cerrahi, dört hastaya amputasyon yapılmıştı. En sık tutulum 23 hasta (%51.1) ile femur alt uç, 10 hasta (%22.2) ile tibia üst uçta görüldü. Üç hastada başvuru anında metastaz vardı. Cerrahi rezeksiyon örnekleri patoloji arşivinden çıkartılarak, p-glikoprotein p170, p53, ısı şok proteini 27 (HSP27), HSP90 ve nm23 proteinlerinin ekspresyonu immünohistokimyasal yöntemlerle incelendi. Bu proteinlerin prognoz ve sağkalım üzerindeki etkileri Kaplan-Meier yöntemi ile değerlendirildi. Hastaların ortalama takip süresi 49.7 ay (dağılım 6-185 ay) idi. Sonuçlar: Başvuru anında metastaz saptanan üç hasta beş yıl içinde akciğer metastazından yaşamını yitirdi. Yirmi dokuz hastada metastaz gelişti. Beş ve 10 yıllık genel sağkalım oranları sırasıyla %60 ve %43 bulundu. Hastalıksız sağkalım oranı ise beş yıl için %41, 10 yıl için %24 idi. Metastaz gelişen hastalarda beş yıllık sağkalım oranı %29 idi. Klinik faktörler içinde, sağkalımı anlamlı etkileyen sadece başvuru anında metastaz varlığı idi (p=0.044). p53 proteininin pozitif ve negatif ekspresyonları arasında beş yıl ve on yıllık sağkalım oranları açısından anlamlı fark görülürken (p=0.04), incelenen diğer proteinler sağkalımla ilişkili bulunmadı. Çıkarımlar: p53 ekspresyonu ile sağkalım arasındaki ilişki, p53’ün osteosarkomda prognostik gösterge olarak kullanılabileceğini düşündürmektedir.

Anahtar Kelimeler:

Sağkalım analizi, Osteosarkom, Neoplazm proteinleri, Tümör belirteci, biyolojik, Tümör baskılayıcı protein p53, İlaç direnci, neoplazm, İmmünohistokimya, Seyir, Kaplan-Meier tahmini

The effect of resistance-related proteins on the prognosis and survival of patients with osteosarcoma: An immunohistochemical analysis

Objectives: Despite the developments in chemotherapy protocols, improvement in the survival rates of osteosarcoma has been limited. We evaluated the effect of certain prognosis-related proteins on survival of patients with osteosarcoma. Methods: Data from 45 patients (24 males, 21 females) who were treated and followed-up for osteosarcoma were reviewed. Following neoadjuvant chemotherapy, 41 patients underwent extremity saving surgery, and four patients underwent amputation. The most frequent localization was the lower end of the femur (n=23, 51.1%), followed by the upper end of the tibia (n=10, 22.2%). Three patients had metastasis on admission. Surgical resection samples were retrieved from the pathology archive and analyzed immunohistochemically for the expression of p-glycoprotein p170, p53, heat-shock protein 27 (HSP27), HSP90, and nm23. The effect of these proteins on prognosis and survival was assessed with survival analysis using the Kaplan-Meier method. The mean follow-up was 49.7 months (range 6 to 185 months). Results: Three patients with metastasis on admission died within five years due to pulmonary metastasis. New metastases developed in 29 patients. Total 5-year and 10-year survival rates were 60% and 43%, respectively. The corresponding diseasefree survival rates were 41% and 24%. Five-year survival was 29% in patients who developed metastasis. Among clinical factors, survival was influenced only by the presence of metastasis on admission (p=0.044). Five-year and 10-year survival rates were significantly different between patients with and without p53 positivity (p=0.04), while the other proteins were not significantly associated with survival. Conclusion: Our data suggest that p53 may be used as a prognostic marker in osteosarcoma due to its significant association with survival.

Keywords:

Survival Analysis, Osteosarcoma, Neoplasm Proteins, Tumor Markers, Biological, Tumor Suppressor Protein p53, Drug Resistance, Neoplasm, Immunohistochemistry, Prognosis, Kaplan-Meier Estimate,

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1. Chou AJ, Gorlick R. Chemotherapy resistance in osteosarcoma: current challenges and future directions. Expert Rev Anticancer Ther 2006;6:1075-85.
2. Davidoff AM, Iglehart JD, Marks JR. Immune response to p53 is dependent upon p53/HSP70 complexes in breast cancers. Proc Natl Acad Sci U S A 1992;89:3439-42.
3. Greenblatt MS, Bennett WP, Hollstein M, Harris CC. Mutations in the p53 tumor suppressor gene: clues to cancer etiology and molecular pathogenesis. Cancer Res 1994;54:4855-78.
4. Miller CW, Aslo A, Won A, Tan M, Lampkin B, Koeffler HP. Alterations of the p53, Rb and MDM2 genes in osteosarcoma. J Cancer Res Clin Oncol 1996;122:559-65.
5. Trieb K, Kotz R. Proteins expressed in osteosarcoma and serum levels as prognostic factors. Int J Biochem Cell Biol 2001;33:11-7.
6. Wadayama B, Toguchida J, Yamaguchi T, Sasaki MS, Yamamuro T. p53 expression and its relationship to DNA alterations in bone and soft tissue sarcomas. Br J Cancer 1993;68:1134-9.
7. Love S, King RJ. A 27 kDa heat shock protein that has anomalous prognostic powers in early and advanced breast cancer. Br J Cancer 1994;69:743-8.
8. Trieb K, Gerth R, Holzer G, Grohs JG, Berger P, Kotz R. Antibodies to heat shock protein 90 in osteosarcoma patients correlate with response to neoadjuvant chemotherapy. Br J Cancer 2000;82:85-7.
9. Serra M, Maurici D, Scotlandi K, Barbanti-Brodano G, Manara MC, Benini S, et al. Relationship between P-glycoprotein expression and p53 status in high-grade osteosarcoma. Int J Oncol 1999;14:301-7.
10. Serra M, Scotlandi K, Manara MC, Maurici D, Benini S, Sarti M, et al. Analysis of P-glycoprotein expression in osteosarcoma. Eur J Cancer 1995;31A:1998-2002.
11. MacDonald NJ, de la Rosa A, Steeg PS. The potential roles of nm23 in cancer metastasis and cellular differentiation. Eur J Cancer 1995;31A:1096-100.
12. Oda Y, Naka T, Takeshita M, Iwamoto Y, Tsuneyoshi M. Comparison of histological changes and changes in nm23 and c-MET expression between primary and metastatic sites in osteosarcoma: a clinicopathologic and immunohistochemical study. Hum Pathol 2000;31:709-16.
13. Ueda Y, Dockhorn-Dworniczak B, Blasius S, Mellin W, Wuisman P, Böcker W, et al. Analysis of mutant P53 protein in osteosarcomas and other malignant and benign lesions of bone. J Cancer Res Clin Oncol 1993;119:172-8.
14. Uozaki H, Horiuchi H, Ishida T, Iijima T, Imamura T, Machinami R. Overexpression of resistance-related proteins (metallothioneins, glutathione-S-transferase pi, heat shock protein 27, and lung resistance-related protein) in osteosarcoma. Relationship with poor prognosis. Cancer 1997;79:2336-44.
15. Bacci G, Ferrari S, Bertoni F, Ruggieri P, Picci P, Longhi A, et al. Long-term outcome for patients with nonmetastatic osteosarcoma of the extremity treated at the Istituto Ortopedico Rizzoli according to the Istituto Ortopedico Rizzoli/osteosarcoma-2 protocol: an updated report. J Clin Oncol 2000;18:4016-27.
16. Bielack SS, Kempf-Bielack B, Delling G, Exner GU, Flege S, Helmke K, et al. Prognostic factors in highgrade osteosarcoma of the extremities or trunk: an analysis of 1,702 patients treated on neoadjuvant cooperative osteosarcoma study group protocols. J Clin Oncol 2002; 20:776-90.
17. Meyers PA, Gorlick R, Heller G, Casper E, Lane J, Huvos AG, et al. Intensification of preoperative chemotherapy for osteogenic sarcoma: results of the Memorial Sloan-Kettering (T12) protocol. J Clin Oncol 1998;16:2452-8.
18. Özger H, Eralp L, Atalar AC, Toker B, Ayan İ, Kebudi R, et al. Survival analysis and the effects of prognostic factors in patients treated for osteosarcoma. [Article in Turkish] Acta Orthop Traumatol Turc 2007;41:211-9.
19. Petrilli AS, de Camargo B, Filho VO, Bruniera P, Brunetto AL, Jesus-Garcia R, et al. Results of the Brazilian Osteosarcoma Treatment Group Studies III and IV: prognostic factors and impact on survival. J Clin Oncol 2006; 24:1161-8.
20. Winkler K, Beron G, Kotz R, Salzer-Kuntschik M, Beck J, Beck W, et al. Neoadjuvant chemotherapy for osteogenic sarcoma: results of a Cooperative German/Austrian study. J Clin Oncol 1984;2:617-24.
21. Scully SP, Ghert MA, Zurakowski D, Thompson RC, Gebhardt MC. Pathologic fracture in osteosarcoma : prognostic importance and treatment implications. J Bone Joint Surg [Am] 2002;84:49-57.
22. Pápai Z, Féja CN, Hanna EN, Sztán M, Oláh E, Szendrôi M. P53 Overexpression as an indicator of overall survival and response to treatment in osteosarcomas. Pathol Oncol Res 1997;3:15-9.
23. Honoki K, Tsutsumi M, Miyauchi Y, Mii Y, Tsujiuchi T, Morishita T, et al. Increased expression of nucleoside diphosphate kinase/nm23 and c-Ha-ras mRNA is associated with spontaneous lung metastasis in rat-transplantable osteosarcomas. Cancer Res 1993;53:5038-42.
24. Schneider J, Jimenez E, Marenbach K, Marx D, Meden H. Co-expression of the MDR1 gene and HSP27 in human ovarian cancer. Anticancer Res 1998;18:2967-71.
25. Uozaki H, Ishida T, Kakiuchi C, Horiuchi H, Gotoh T, Iijima T, et al. Expression of heat shock proteins in osteosarcoma and its relationship to prognosis. Pathol Res Pract 2000;196:665-73.
26. Chan HS, Grogan TM, Haddad G, DeBoer G, Ling V. P-glycoprotein expression: critical determinant in the response to osteosarcoma chemotherapy. J Natl Cancer Inst 1997;89:1706-15.