some novel insights on hpv16 related cervical cancer pathogenesis based on analyses of lcr methylation, viral load, e7 and e2e4 expressions一些新奇的见解hpv16相关宫颈癌发病机制基于电感电容电阻测量甲基化分析,病毒载量、e7 e2e4表达式.pdfVIP

Some Novel Insights on HPV16 Related Cervical Cancer Pathogenesis Based on Analyses of LCR Methylation, Viral Load, E7 and E2/E4 Expressions Damayanti Das Ghosh1., Bornali Bhattacharjee1.¤, Shrinka Sen4., Laikangbam Premi1., 1 2 3 4 Indranil Mukhopadhyay , Rahul Roy Chowdhury , Sudipta Roy , Sharmila Sengupta * 1 Human Genetics Unit, Indian Statistical Institute, Kolkata, India, 2 Department of Gynecology, Saroj Gupta Cancer Centre and Research Institute, Kolkata, India, 3 Department of Pathology, Suraksha Diagnostics Private Limited, Kolkata, India, 4 National Institute of Biomedical Genomics, Kalyani, Dist. Nadia, West Bengal, India Abstract This study was undertaken to decipher the interdependent roles of (i) methylation within E2 binding site I and II (E2BS-I/II) and replication origin (nt 7862) in the long control region (LCR), (ii) expression of viral oncogene E7, (iii) expression of the transcript (E7-E1‘E4) that encodes E2 repressor protein and (iv) viral load, in human papillomavirus 16 (HPV16) related cervical cancer (CaCx) pathogenesis. The results revealed over-representation (p,0.001) of methylation at nucleotide 58 of E2BS-I among E2-intact CaCx cases compared to E2-disrupted cases. Bisulphite sequencing of LCR revealed overrepresentation of methylation at nucleotide 58 or other CpGs in E2BS-I/II, among E2-intact cases than E2-disrupted cases and lack of methylation at replication origin in case of both. The viral transcript (E7-E1‘E4) that produces the repressor E2 was analyzed by APOT (amplification of papillomavirus oncogenic transcript)-coupled-quantitative-RT-PCR (of E7 and E4 genes) to distinguish episomal