Brn-3a Expression In Cervical Lesion And Its Relationship With Different HPV Subtypes

Objective The aim of this study is to understand the expression of cellular transcription factor Brn-3a in cervical lesions caused by various subtypes of HPV,and analyze Brn-3a's role in HPV induced cervical carcinogenesis,utimately explore its potential as a diagnostic and treatment marker.Methods Cervical smears from 97 women with cervical lesions attending the Obstetrics & Gynecology Hospital affiliated to Fudan University were collected according to our enrollment requirements,among which 5 groups were divided:1) chronic cervicitis(27 cases);2) cervcial condyloma and CIN1(18 cases);3) CIN2,CIN3(27 cases);4) cervical invasive cancer(15 cases);5) CIN2 and CIN3 patients 3 months after LEEP treatment(10 cases). Patients'consent having been acquired,samples were collected with a TCT brush before treatment(except the CIN2,3 after LEEP group).The mRNA levels of Brn-3a were measured with real-time RT-PCR and the data obtained were used to establish the relationship between Brn-3a and the histopathology.The different HPV types each infected were tested with PCR and pre-designed flow-through hybridization gene chips,by which high risk subtypes:16,18,31,33,35,39,45,51,52,56,58,59,66;low risk subtypes: 6,11,42,43,44,53,68 and cp8304 can be identified.Patients'diagnosises and treatment processes were closely followed up,and all the final diagnosises were made from decisive histopathology.Data were analyzed with Stata 8.0.The Wilcoxon,Kruskal-Wallis H and Spearman tests were used to compare the related parameters,while P value＜0.05 indicates the existence of statistically significant difference.Results①Among all the samples,the most frequently infected HPV subtypes were sequentially:HPV 16(20.62%),HPV 58(13.40%),HPV 52 (9.28%) and HPV 53(9.28%);in CIN2,3 lesions,the HPV subtypes infected were mainly:HPV 16(44.44%),HPV 58(22.22%),HPV 52(14.81%) and HPV 31 (14.81%);while in cases diagnosed as cervical condyloma and CIN1 were mostly:HPV58,16(16.67%),HPV52,53,68(12.50%),HPV11(8.33%)and HPV33,6,59,18,CP8304(4.17%).②The Brn-3a Mean Qty of the predefined 5 groups by real time PCR were 0.0138656±0.0307035,0.6680568±2.380923, 12.3093±23.76736,29.23117±37.2152 and 0.268678±0.687342, respectively.The quantified level of Brn-3a corresponded to the severity of the cervical lesions,and its value in cervical cancer group differred significantly from the chronic cervicitis group(P＝0.0000,＜0.05), cervical condyloma and CIN1(P＝0.0000,＜0.05),CIN2,3(P＝0.0005,＜0.05) and CIN2,CIN3 post- LEEP group(P＝0.0003,＜0.05).In cervicitis, difference of Brn-3a expression in the groups of HPV- and HPV+ was not statistically significant(P＝1.000,＞0.05).Brn-3a expression in the CIN2,3 after LEEP group was statistically higher than the chronic cervicitis group(P＝0.0045,＜0.05) and lower than the 3rd group(P＝0.0004,＜0.05);but its discrepancy with the 2nd group was not statistically significant (P＝0.5020,＞0.05);③No statistically significant difference was found in Brn-3a level between diverse HPV subtypes in the same degree of cervical lesions,or between high risk and low risk subtypes of HPV(P＝0.5815,＞0.05) when analyzed as a whole in≤CIN1 lesions which was the only group infected by both high and low risk types of HPV.While in different degrees of cervical lesions infected with the same subtype of HPV,the expression of Brn-3a varied significantly.Conclusion①The expression of Brn-3a relates positively to histopathology,and does not vary significantly according to different HPV subtypes in the same degree of cervical lesion.②It was significantly lower in the group of CIN2,3 after LEEP treatment than the CIN2,3 group; which may be a reflection of the treatment and patients'recovery.③Brn-3a may play a critical role in different types of HPV induced carcinogenesis,and can be used as an independent marker for cervical lesions.