Immunohistochemical Study and in-situ hybridization for detection of humanpapillomavirus (HPV ) in Uterine Cervical Carcinoma

Cervical cancer is the second most common life-threatening cancer among women worldwide. The human papilloma virus (HPV) has been definitely linked as the main etiologic agent for cervical carcinoma. To date, there are more than 200 known HPV types, and they are generally classified according to their potential to induce malignant transformation. HPVs 16,18, 31, 33, 35, 39, 45, 50, 51,53, 55, 56, 58, 59, 64 and 68 are considered “high risk” types because they are detectable in carcinomas and dysplasias and low risk types (non-oncogenic:6,11,40,42,43,44,54,61,72,73,81). Immunohistochemistry was performed for detection of HPV proteins on cervical tissues. Chromogenic in situ hybridization was performed to detect HPV DNA and determine high-risk HPV on cervical samples. Aim of the Study This study was designed to assess rate of detection and prevalence of HPV in cervical carcinoma hysterectomized patients through both immunohistological and chromogenic in-situ hybridization techniques and to identify and to differentiate histopathologically the high risk genotype of HPV (16, 18 ) from low risk genotype ( 6,11 ). Materials and Methods A total 75 cervical tissue samples of female patients, who had undergone hysterectomy or submitted for punch biopsy from their cervices, were included in this study, that conducted in the Department of Pathology, Faculty of Medicine, Kufa University from December 2010 through June 2011. Forty three patients with cervical carcinoma (57.3%) (40 patients with squamous cell carcinoma and 3 with adenocarcinoma) and the remaining 32(42.7%) normal cervical tissue were considered as control group involved in this study. All the cases were collected from Al-Sadder Teaching Hospital and some of the private laboratories in Najaf. Their ages were ranging between 28-65 years with a mean age of 46.02 years. Labeled Streptavidin -Biotin (LSAB+) method and GenPoint TM Tyramide Signal Amplification System were employed for immunohistochemical and chromogenic in situ hybridization detection of HPV respectively. Statistical analysis was achieved by Chi-square test and correlation-regression test at p value< 0.05 and ? 0.3 respectively. Results The immunohistochemistry detection rate of HPV 6/11/16/18/ 31/33/ 35/45/ 51/52 was reported in 18 (41.9%) out of 43 cases of cervical carcinoma. Chromogenic in situ hybridization detection rate of HPV 6/ 11/16/ 18/ 31/ 33/ 35/ 45/ 51/52 was reported in 15 (34.9%) out of 43 cases of cervical carcinoma .However Chromogenic in situ hybridization detection rate of high risk genotypes of HPV(16, 18, 31, 33, 35, 39, 45, 51, 52, 56, 58, 59, and 68) was reported in 17 (39.5%) out of 43 cases of cervical carcinoma. All normal control and adenocarcinoma cases showed no specific signals for HPV DNA and protein . There was a significant difference of HPV immunoexpression among types of cervical tissue and age of the patients . However, HPV was positively correlated with tumor grade and size(T) of tumor. A significant correlation was found between both detection methods of HPVs (immunohistochemistry and CISH techniques) in the cases of cervical carcinoma. Conclusions and recommendations From the above results we concluded that HPV over-expression plays an important role in pathogenesis of cervical carcinoma . So incorporation of this marker with other pathological parameters will more accurately predict clinical outcome and determine the effects of anti cancer therapy.