Objectives: This study investigates the potential value of Raf kinase inhibitor protein (RKIP) as a marker of normal squamous cells in ThinPrep slides. RKIP was evaluated for its ability to distinguish between normal and abnormal cervical samples in the context of human papillomavirus (HPV) infections. Study Design: A total of 316 ThinPrep samples were taken from women with normal and abnormal cervices. ThinPrep slides were Papanicolaou stained and reported. Residual samples were used for RKIP immunostaining and HPV PCR-based sequencing. Results: RKIP expression was seen in both nuclei and cytoplasm in 83.7% of samples. RKIP expression was highest (84.6%) in samples with a diagnosis of high-grade squamous intraepithelial lesion (HSIL) or worse; expression was lower in low-grade squamous intraepithelial lesions (73%) and was lowest in samples with normal cytology (p = 0.0023). A total of 74% of HPV-infected ThinPrep samples were immunopositive, and 67% of samples that did not harbor HPV were also immunopositive (p = 0.414). Sensitivity and specificity of RKIP were 84.6 and 34.6%, respectively, for the detection of samples with HSIL or worse. Conclusions: This study showed that RKIP expression may be of some value as a marker for abnormal cervical cells. Combined RKIP expression and HPV testing could improve the identification of samples with abnormal cytology.

Keywords:
Raf kinase inhibitor protein expression, Human papillomavirus infection, ThinPrep cervical samples
1.
de Sanjose S, Diaz M, Castellsague X, Clifford G, Bruni L, Manuz N, Bosch FX: Worldwide prevalence and genotype distribution of cervical human papillomavirus DNA in women with normal cytology: a meta-analysis. Lancet Infect Dis 2007;7:453-459.
2.
Schiffman M, Castle PE: Human papillomavirus: epidemiology and public health. Arch Pathol Lab Med 2003;127:930-934.
3.
Munoz N, Bosch X, de Sanjose, Herrero R, Castellsague X, Shah K, Snijders P, Meijer C; International Agency for Research on Cancer Multicenter Cervical Cancer Study Group: Epidemiologic classification of human papillomavirus types associated with cervical cancer. N Engl J Med 2003;348:518-527.
4.
Schiffman M, Castle PE, Jeronimo J, Rodriguez AC, Wacholder S: Human papillomavirus and cervical cancer. Lancet 2007;370:890-897.
5.
Al-Mulla F, Bitar MS, Taqi Z, Rath O, Kolch W: RAF kinase inhibitory protein (RKIP) modulates cell cycle kinetics and motility. Mol Biosyst 2011;7:928-941.
6.
Park S, Yeung ML, Beach S, Shields JM, Yeung KC: RKIP down regulates B-Raf kinase activity in melanoma cancer cells. Oncogene 2005;24:3535-3540.
7.
Hagan S, Al-Mulla F, Mallon E, Oien K, Ferrier R, Gusterson B, Garcia JJ, Kolch W: Reduction of Raf-1 kinase inhibitor protein (RKIP) expression in breast cancer metastases. Clin Cancer Res 2005;11:7392-7397.
8.
Al-Mulla F, Hagan S, Behbehani AI, Bitar MS, George SS, Going JJ, Curto García JJ, Scott L, Fyfe N, Murray GI, Kolch W: Raf kinase inhibitor protein expression in a survival analysis of colorectal cancer patients. J Clin Oncol 2006;24:5672-5679.
9.
Lee HC, Tian B, Sedivy JM, Wands JR, Kim M: Loss of Raf kinase inhibitor protein promotes cell proliferation and migration of human hepatoma cells. Gastroenterology 2006;131:1208-1217.
10.
Schuierer MM, Bataille F, Hagan S, Kolch W, Bosserhoff AK: Reduction in Raf kinase inhibitor protein expression is associated with increased Ras extracellular signal-regulated kinase signaling in melanoma cell lines. Cancer Res 2004;64:5186-5192.
11.
Wang J, Yang YH, Wang AQ, Yao B, Xie G, Feng G, Zhang Y, Cheng ZS, Hui L, Dai TZ, Du XB, Wang D: Immunohistochemical detection of the Raf kinase inhibitor protein in nonneoplastic gastric tissue and gastric cancer tissue. Med Oncol 2010;27:219−223.
12.
Li HZ, Wang Y, Gao Y, Shao J, Zhao XL, Deng WM, Liu YX, Yang J, Yao Z: Effects of Raf kinase inhibitor protein expression on metastasis and progression of human epithelial ovarian cancer. Mol Cancer Res 2008;6:917−928.
13.
Zhang L, Fu Z, Binkley C, Giordano T, Burant CF, Logsdon CD, Simeone DM: Raf kinase inhibitory protein inhibits β-cell proliferation. Surgery 2004;136:708-715.
14.
Hu CJ, Zhou L, Zhang J, Huang C, Zhang GM: Immunohistochemical detection of Raf kinase inhibitor protein in normal cervical tissue and cervical cancer tissue. J Int Med Res 2011;39:229-237.
15.
Bibbo M, DeCecco J, Kovatich AJ: P16INK4A as an adjunct test in liquid-based cytology. Anal Quant Cytol Histol 2003;25:8-11.
16.
Sahebali S, Depuydt CE, Segers K, Vereecken AJ, Van Marck E, Bogers JJ: Ki-67 immunocytochemistry in liquid based cervical cytology: useful as an adjunctive tool? J Clin Pathol 2003;56:681-686.
17.
Al-Awadhi R, Van Noorden S, Coleman DV: Evaluation of NMP179 for the detection of squamous intraepithelial neoplasia in ThinPrep cervical slides using a combination of double immunostaining and morphometric methods of analysis. Diagn Cytopathol 2004;31:135-140.
18.
Pinto AP, Degen M, Villa LL, Cibas ES: Immunomarkers in gynecologic cytology: the search for the ideal ‘biomolecular Papanicolaou test'. Acta Cytol 2012;56:109-121.
19.
Sahasrabuddhe VV, Luhn P, Wentzensen N: Human papillomavirus and cervical cancer: biomarkers for improved prevention efforts. Future Microbiol 2011;6:1083-1098.
20.
National Cancer Institute (2001). Bethesda System 2001. Available at: http://www.bethesda2001.cancer.gov (accessed on June 11, 2010).
21.
Kounelis S, Kapranos N, Kouri E, Coppola D, Papadaki H, Jones MW: Immunohistochemical profile of endometrial adenocarcinoma: a study of 61 cases and review of the literature. Mod Pathol 2000;13:379-388.
22.
Tashiro H, Isacson C, Levine R, Kurman RJ, Cho KR, Hedrick L: p53 gene mutations are common in uterine serous carcinoma and occur early in their pathogenesis. Am J Pathol 1997;150:177-185.
23.
Al-Awadhi R, Chehadeh W, Kapila K: Prevalence of human papillomavirus among women with normal cervical cytology in Kuwait. J Med Virol 2011;83:453-460.
24.
Al-Awadhi R, Chehadeh W, Jaragh M, Al-Shaheen A, Sharma P, Kapila K: Distribution of human papillomavirus among women with abnormal cervical cytology in Kuwait. Diagn Cytopathol 2013;41:107-114.
25.
Lum A, Le Marchand L: A simple mouthwash method for obtaining genomic DNA in molecular epidemiological studies. Cancer Epidemiol Biomarkers Prev 1998;7:719-724.
26.
Gravitt PE, Peyton CL, Apple RJ, Wheeler CM: Genotyping of 27 human papillomavirus types by using L1 consensus PCR products by a single hybridization reverse line blot detection method. J Clin Microbiol 1998;36:3020-3027.
27.
de Roda Husman AM, Walboomers JMM, van den Brule AJ, Meijer CJ, Snijders PJ: The use of general primers GP5 and GP6 elongated at their 3′ with adjacent highly conserved sequences improves human papillomavirus detection by PCR. J Gen Virol 1995;76:1057-1062.
28.
Speich N, Schmitt C, Bollmann R, Bollmann M: Human papillomavirus (HPV) study of 2916 cytological samples by PCR and DNA sequencing: genotype spectrum of patients from the West German area. J Med Microbiol 2004;53:125-128.
29.
Muñoz N, Castellsagué X, de González AB, Gissmann L: Chapter 1: HPV in the etiology of human cancer. Vaccine 2006;24(suppl 3):S3/1-S3/10.
30.
Biewenga P, Buist MR, Moerland PD, van Themaat EV, van Kampen AHC, Kate F, Baas F: Gene expression in early stage cervical cancer. Gynecol Oncol 2008;108:520-526.
31.
Keller ET, Fu Z, Brennan M: The biology of a prostate cancer metastasis suppressor protein: Raf kinase inhibitor protein. J Cell Biochem 2005;94:273-278.
32.
Klysik J, Theroux S, Sedivy J, Moffit J, Boekelheide K: Signaling crossroads: the function of Raf kinase inhibitory protein in cancer, the central nervous system and reproduction. Cell Signal 2008;20:1-9.
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2013
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