Meme Kanserlerinde HtrA2 Gen İfadesinin Analizi

Abstract

Amaç: Bu çalışmada meme kanserli hastaların tümörlü dokularında ve çevresindeki normal dokularında, High temprature requirement factor A2 HtrA2 geninin hem mRNA hem de protein ifade seviyelerinin tespitini amaçladık.Materyal ve metod: Cerrahi olarak 53 hastadan alınan dokularda mRNA ifade seviyelerinin tespiti qRTPCR ile taze dokularda çalışıldı. Aynı hastalara ait parafine gömülü dokularda ise immünohistokimyasal yöntemle protein tespiti yapıldı. HtrA2 ifade seviyeleri ile hastaların klinik ve patolojik özellikleri arasındaki ilişkiler analiz edildi.Bulgular: Tümörlü dokulardaki HtrA2 geni mRNA ifade seviyeleri, bitişik normal dokulardakiler ile karşılaştırıldığında istatistiksel olarak anlamlı bir fark bulunmadı p=0.074 . HtrA2 proteini, %90.6 hastanın 48/53 tümör dokusunda pozitif olarak tespit edilirken, %9.4 hastanın 5/53 tümör dokusunda ise tespit edilmedi. Tümörlü dokulardaki HtrA2 protein ifadesi, bitişik normal dokulardakiler ile karşılaştırıldığında istatistiksel olarak anlamlı bir fark bulundu p=0.007 .Sonuç: Meme tümör dokularında, HtrA2'nin farklı ifade seviyelerinde tespit edilmesi ile apoptotik yolağı etkinleştirebileceği söz konusu olabilir. Bu nedenle meme kanserinde özellikle IAP'lerin ifade seviyelerinin de tespit edilmesinin önemli olacağını düşünmekteyiz

Keywords

• HtrA2,qRT-PCR,immunohistokimya,meme kanseri,apoptozis

Analysis of HtrA2 Gene Expression in Breast Cancers

Abstract

Background: The purposes of this study were to measure both the protein and mRNA expression levels of HtrA2 gene in human with breast cancer tissues and their adjacent normal breast tissues. Methods: mRNA expression levels in tissues taken surgically from 53 patients detection of fresh tissues were studied by qRT-PCR. The protein detection is done in paraffin-embedded tissues of the same patients through immunohistochemical method. The relationships between patients clinical, pathologic features and HtrA2 expression levels are analyzed. Results: Expression levels of HtrA2 mRNA in tumor tissues was not significantly different statically compared with adjacent normal tissues p=0.074 . While HtrA2 protein level is to identified positively in tumor tissue of patients 90.6% 48/53 , there is no detection of HtrA2 protein expression in tumor tissues of patients 9.4% 5/53 . Expression of HtrA2 protein in tumor tissues was significantly different statically compared with adjacent normal tissues p=0.007 .Conclusions: It is possible that apoptotic patway may be activated by the different expression levels of HtrA2 in breast tumor tissues. Therefore, we concluded that it is particularly important to detect the expression levels of IAPs in breast cancer

Keywords

• HtrA2,qRT-PCR,immunohistochemistry,breast cancer,apoptosis

References

1. ) Kesteloot ECH, Zhang J. Differences in breast cancer mortality worldwide: unsolved problems. European Journal of Cancer Prevention 2006; 15(5): 416-423.
2. )w w w. w o r l d w p i d w b r e a s t c a n c e r. ( E r i ş i m tarihi:28.02.2012)
3. )Türkiye Cumhuriyeti Sağlık Bakanlığı Organlara, cinsiyete ve yaşa göre kanser sıklığının dağılımı ve kadınlarda en sık görülen 10 kanser, Kanserle Savaş Daire Başkanlığı 2006; http://www.saglik.gov.tr./TR (Erişim tarihi: 01.03.2012)
4. ) Consen SD, Grushko TA, Olopade OI. Cancer of the Breast. In: Cancer Principles and Practice of oncology. Devita Jr VT, Lawrance TS, Rosenberg SA (Eds). 8 th Ed. 2008; p.1595-1605.
5. ) Lipinska B, Sharma S, Georgopoulos C. Sequence analysis and regulation of the HtrA gene of Escherichia coli: a sigma 32-independent mechanism of heat- inducible transcription. Nucleic Acids Res 1988; (21):10053–10067.
6. ) Strauch KL, Beckwith J. An escherichia coli mutation preventing degradation of abnormal periplasmic p r o t e i n s . P r o c N a t l A c a d S c i U S A ;85(5):1576–1580.
7. ) Zumbrunn J, Trueb B. Primary structure of a putative serine protease specific for IGF-binding proteins. FEBS Lett. 1996;398(2-3) :187–192.
8. ) Gray CW, Ward RV, Karran E, Turconi S, Rowles A, Viglienghi D, Southan C, Barton A, Fantom KG, West A, Savopoulos J, Hassan NJ, Clinkenbeard H, Hanning C, Amegadzie B, Davis JB, Dingwall C, Livi GP, Creasy CL. Characterization of human HtrA2, a novel serine protease involved in the mammalian cellular stress response. Eur J Biochem 2000; 267(18):5699–5710.

9. ) Nie GY, Hampton A, LI Y, Fındlay JK, Salamonsen LA. Identification and cloning of two isoforms of human high-temperature requirement factor A3 (HtrA3), characterization of its genomic structure and comparison of its tissue distribution with HtrA1 and HtrA2. Biochem. J 2003; 371(1): 39-48.
10. ) Suzuki Y, Imai Y, Nakayama H, Takahashi K, Takio K, Takahashi R. A serine protease HtrA2, is released from the mitochondria and interacts with XIAP, inducing cell death. Mol. Cell 2001;8(3): 613–621
11. ) Martins LM, Morrison A, Klupsch K, Fedele V, Moisoi N, Teismann P, Abuin A, Grau E, Geppert M, Livi GP, Creasy CL, Martin A, Hargreaves I, Heales SJ, Okada H, Brandner S, Schulz JB, Mak T, Downward J. Neuroprotective role of the reaper-related serine protease HtrA2/Omi revealed by targeted deletion in mice. Mol Cell Biol 2004;24(22):9848–9862.
12. ) Su D, Su Z, Wang J, Yang S, Ma J. UCF-101, a novel Omi/HtrA2 inhibitor, protects against cerebral ischemia/reperfusion injury in rats. Anat Rec (Hoboken) ;292: 854–861. ) Saelens X, Festjens N, Vande Walle L, van Gurp M, van Loo G, Vandenabeele P. Toxic proteins released from mitochondria in cell death. Oncogene 2004; 23(16): –2874.
13. ) Cole P, Rodu B. Descriptive Epidemiology: Cancer statistics, In: Cancer Principles Practice of Oncology, Eds: Devita VT, Hellman S, Rosenkerg SA. 6 th ed. p.228-234. ) Zurawa JD, Kobiela J, Stefaniak T, Wozniak A, Narkiewicz J, Wozniak M, Limon J, Lipinska B. Changes in expression of serine proteases HtrA1 and HtrA2 during estrogen-induced oxidative stress and nephrocarcinogenesis in male Syrian hamster. Acta Biochimica Polonica. 2008;55(1):9-19.
14. ) Lee SH, Lee JW, Kim HS, Kim SY, Park WS, Kim SH, Lee JY, Yoo NJ. Immunohistochemical analysis of Omi/HtrA2 expression in stomach cancer. APMIS 2003; (5): 586–590.
15. ) Narkiewicz J, Klasa MD, ZurawaJD, SkorkoGJ, Emerich J, Lipinska B. Changes in mRNA and protein levels of human HtrA1, HtrA2 and HtrA3 in ovarian cancer. Clinical Biochemistry 2008;41(7-8) 561–569.
16. ) Narkiewics J, Lapinska-Szumczyk S, ZurawaJD, SkorkoGJ, Emerich J, Lipinska B. Expression ofhuman HtrA1, HtrA2, HtrA3 and TGF-ß1 genes in primary endometrial cancer. Oncology Reports 2009; 21(6): 1529
17. ) Hu XY, Xu YM, Chen XC, Ping H, Chen ZH, Zeng FQ. Immunohistochemical analysis of Omi/HtrA2 expression in prostate cancer and benign prostatic hyperplasia. APMIS 2006;114(12): 893–898.
18. ) Chen HL, Chen CQ, Ma JP et al. Association of Omi/HtrA2 expression and prognosis in patients with gastric carcinoma. Zhonghua Wei Chang Wai Ke Za Zhi.2010 Oct;13(10):766-9.
19. ) Bhuiyan S, Fukunaga K. Activation of HtrA2, a Mitochondrial Serine Protease Mediates Apoptosis: Current Knowledge on HtrA2 Mediated Myocardial Ischemia/Reperfusion Injury. Cardiovasc Ther 2008; (3): 224–232.
20. ) Schimmer AD. Inhibitor of apoptosis proteins: translating basic knowledge into clinical practice. Cancer Res 2004; 64(20):7183–7190.