The Expression Of P16, ΔNP63, HPV16 E7 And Brn-3a In Thinprep Liquid-Based PAP Smears And The Relationship With Cervical Premalignant Lesions

Object: To examine the potential of p16, ?NP63, HPV16 E7 and Brn-3a as a biomarker for cervical premalignant lesions in ThinPrep smears. And search for the identification of specific biomarkers for dysplastic epithelial cells of the cervix as a new examine ways to reduce the failure of the Pap test. Methods: 1.Random sample 71 patients who were doubtfully diagnose as CIN to be examined ThinPrep smears and to be done cervical biopsy under the colpomicroscope. Using streptavidin peroxidase (SP) method, Immunocytochemical analysis of p16 and ?NP63 expression was performed on 71 ThinPrep smears. 2.RT-PCR was used to detect the mRNA of HPV16 E7 and Brn-3a in 71 ThinPrep liquid-based smears specimens. Results: 1.According to the 2001 Bethesda System, the cytologic diagnoses of 71 cases are including 13 cases for Negative for Intraepithelial Lesion or Malignancy (NILM), 17 cases for atypical squamous cells (ASC), 19 cases for low grade squamous intraepithelial lesion (LSIL) and 22 cases for high grade squamous intraepithelial lesion (HSIL). The pathologic diagnoses of 71 cases are including 21 cases for cervical inflammation, 19 cases for cervical intraepithelial neoplasia1 (CINⅠ ), 10 cases for CINⅡ and 21 cases for CINⅢ . 2.In ThinPrep smears, the positive of P16 is found in the cytoplast of dysplastic epithelial cells of the cervix and adenocervical celle. In cytologic diagnose grade, the positive rates of P16 is respectively NILM (15.38%), ASC (54.55%), LSIL (95.45%), HSIL (100%). There are marked diference in both HSIL vs NILM and LSIL vs NILM (P<0.0001) but unmarked diference in both NILM vs ASC and LSIL vs HSIL (P>0.05). In pathologic diagnose grade,the positive rates of it is respectively inflammation(0%),CINⅠ(5.26%),CINⅡ(50%),CINⅢ(90.48%). And there are marked diference between inflammation and CIN (P<0.0001) but unmarked diference among CINs (P>0.05). 3. In ThinPrep smears, ΔNP63 is positive in the dysplastic squamous cells of the cervix and adenocervical cells. In cytologic diagnose, the positive rates of it are NILM(15.38%),ASC(47.06%),LSIL(89.47%),HSIL(95.45%). And there are marked diference in NILM vs LSIL, NILM vs HSIL and ASC vs HSIL (P<0.001) but unmarked diference in NILM vs ASC and LSIL vs HSIL (P>0.05). In pathologic diagnose grades, the positive rates of it are respectively inflammation(4.76%),CINⅠ(84.21%),CINⅡ(100%),CINⅢ(100%). And there are marked diference between inflammation and CINs (P<0.001) but unmarked diference among CINs (P>0.05). 4. In cytologic diagnose grades, the positive rates of HPV16 E7 in ThinPrep samples are respectively NILM(7.69%),ASC(29.41%),LSIL(31.58%),HSIL(59.09%). And there are marked diference between NILM and HSIL (P<0.008) but unmarked diference in NILM vs ASC, NILM vs LSIL and LSIL vs HSIL (P>0.05). In pathologic diagnose grades, the positive rates of it are respectively inflammation(0%),CINⅠ(31.58%),CINⅡ(60%),CINⅢ(61.90%). And there are marked diference in CINⅡ vs inflammation and CINⅢ vs inflammation (P<0.001) but unmarked diference among CINs (P>0.05).5. In cytologic diagnose grades, the positive rates of Brn-3a in ThinPrep samples are respectively NILM(7.69%),ASC(23.53%),LSIL(21.05%),HSIL(72.73%). And there are marked difference in HSIL vs NILM, HSIL vs ASC and HSIL vs LSIL (P<0.001) but unmarked diference among NILM, ASC and LSIL (P>0.05). In pathologic diagnose grades, the positive rates of it are respectively inflammation(0%),CINⅠ(5.26%),CINⅡ(50%),CINⅢ(90.48%). And there are marked differences in CINⅢ vs CINⅠ and CIN Ⅲ vs inflammation (P<0.0001) but unmarked diference in CINⅠ vs inflammation and CINⅡ vs CINⅢ (P>0.05). 6. As the one of diagnostic biomarkers of cervical intraepithelial neoplasia, the results of P16 immunostaining are consistent with that of ?NP63 (P<0.05). As the one of biomarkers of cervical intraepithelial neoplasia and activation of HR-HPV's oncogene, the results of HPV16 E7 are consistent with that of Brn-3a (P<0.05). Conclusions: 1. The pattern of overexpression demonstrates the potential use of p16 as a diagnostic...