استخدام الصبغة المناعية p16INK4a والتهجين الموقعي وتاثيرها على فحوصات عنق الرحم الصحيحة والملتبسة == The Use of p16INK4a and In Situ Hybridization and Their Impact on the Healthy and Equivocal Pap Smears of the Cervix
Author name:
ثناء جميل الخشالي
Supervisor name:
ندى صالح امين | هدى مهدي الخطيب
General topic:
Medicine
Specific topic:
Anatomy and Histology and Embryos
Degree:
Doctorate
University:
University of Baghdad - Faculty Of Medicine
Language:
English
University location:
Baghdad
First pages:
19T1621 - p.pdf
Abstract:
Throughout the past thirty years, the perception of cervicalcarcinoma has shifted from that of a mysteriously fatal disease, to one ofthe sexually transmitted, human papillomavirus (HPV) related infection.HPV DNA has been found in almost all cervical invasive and preinvasivecervical neoplasms.Conventional Pap smear, which was established in the early sixtiesreduced effectively the morbidity and mortality related to cervical cancer.Given the lower sensitivity of the Pap cytology test, new diagnosticparameters have been established. The histological features ofpreinvasive cervical neoplasia (CIN1, CIN2, and CIN3) are wellunderstood. However, misinterpretation of the morphological criteriacould lead to significant variability.The aim of this study is to identify women with equivocal and normalPap smear who are at risk for developing cervical cancer through the useof p16INK4a immunostaining and human papillomavirus (HPV) DNAtesting using in situ hybridization (ISH).Materials and MethodsIn order to identify human papillomavirus infection in minorcytological and histological abnormalities, biopsies collected from 60women 25 - 66 years of age with normal cervices and Pap smears (groupI), abnormal cervices and Pap smears (group II) and with cervicalcarcinoma (groupIII) referred to the Colposcopy Clinic in BaghdadTeaching Hospital - Medical City Complex, during the period from June2013 through July 2014. Cytology samples collected with ThinPrep forIVliquid base cytology (LBC), punch biopsy for histopathology formalinfixed and paraffin embedded. In situ hybridization (ISH) and p16INK4aimmunohistochemistry in addition to routine hematoxylin and eosin(H&E) stain were used to evaluate the histological specimens.ResultsThe age, age of marriage and parity of the three groups werestudied in relation to each of ISH and p16INK4a reactions. Agedistribution was highly significant among all age groups; in both ISH andp16 test results GI &GII (p=0.00), GI & GIII (p=0.00), GII &GIII(p=0.00) in both ISH and p16 test results. The age of marriage was notsignificant among the groups in both ISH and p16; GI & GII p (0.93), GI&GIII (0.12), GII &GIII (0.30). The effect of parity was insignificantamong all groups; GI & GII = (p = 0.66), GI & GIII (p = 0.82), GII &GIII (p = 0.79) in both ISH and p16 test resultsp16INK4a immunoreaction : Among group I there was a negativereaction in all the specimens collected. In specimens from group II, therewas a very mild or focal p16INK4a immunoreaction in one specimen(5%), a moderate immunoreactivity in 9 specimens (45%), and a highpositive reactivity in 10 specimens (50%) (p=0.000). Group III showedvery high immunoreaction in all the specimens collected.In situ hybridization (ISH) in group I was mild positive in 4specimens (20%) which showed diffuse signal pattern. A mild positivereaction with diffuse nuclear staining was noticed in 4 (20%) of thecervical specimens. Moderate positive reaction with diffuse staining ofthe nucleus was observed in 8 (40%) of the specimens. The remaining 8(40%) of the specimens showed both diffuse and punctate staining of theVnuclei. Among group III specimens, the nuclei in the epithelial cellsshowed both punctate and diffuse signal patterns.Chi square was done to compare results concerning ISH & variousgroups was highly significant (p= 0.00) ConclusionHPV infection is prevalent and can be missed by the conventional Pap smear. The severity of the disease was directly proportional with the age of women in this study. p16 overexpression was correlated with the positivity of ISH. This may support the use of either, or both, tests to improve the accuracy of cervical intraepithelial neoplasia (CIN) diagnosis, and help in the triage of women with equivocal lesions.
