title: Hypomethylated CpG around the transcription start site enables TERT expression and HPV16 E6 regulates TERT methylation in cervical cancer cells.
abstract: OBJECTIVE
  The human papillomavirus (HPV) oncoprotein, E6, activates telomerase reverse transcriptase (TERT) expression and causes cellular immortalization. It remains unclear whether E6 affects TERT transcription by altering DNA methylation profiles. In this study, we explored the methylation status of the TERT promoter in cervical cancer cell lines and its variations after E6 was silenced by RNAi.
  
  
  METHODS
  Three kinds of cervical cell lines (HPV16 positive: CaSki and SiHa; HPV18 positive: HeLa), were taken to analyze the methylation status of the TERT promoter by methylation-specific polymerase chain reaction (MSP) and bisulfite sequencing (BS). Stealth RNAi was transiently transfected to these cell lines to silence the expression of HPV16/18 E6, and the subsequent changes of TERT mRNA levels and TERT promoter DNA methylation were examined.
  
  
  RESULTS
  Hypomethylation of the DNA around the TERT transcription start site (-156 to +162 bp) was functionally related to its transcription. After transfection with Stealth RNAi, the levels of HPV16/18 E6 and TERT mRNA were greatly decreased. The methylated CpG around the transcription start sites in CaSki and SiHa cells were statistically increased (respectively P=0.016, P=0.000). However, there was no significant difference in HeLa cells (P=0.128).
  
  
  CONCLUSIONS
  Hypomethylated CpG around the transcription start site enables the expression of TERT in cervical cancer cells. Our results show for the first time that HPV16 E6 can promote TERT transcription through demethylating the DNA sequence around the TERT transcription start site in cervical squamous cancer cells.
authors: Department of Obstetrics and Gynecology, Peking University People's Hospital, Beijing 100044, China.
authors: Jing
authors: J
authors: Jiang
authors: Li-Jun
authors: LJ
authors: Zhao
authors: Chao
authors: C
authors: Zhao
authors: Guo
authors: G
authors: Zhang
authors: Yun
authors: Y
authors: Zhao
authors: Jing-Ran
authors: JR
authors: Li
authors: Xiao-Ping
authors: XP
authors: Li
authors: Li-Hui
authors: LH
authors: Wei
mesh_terms: D018899:CpG Islands; D019175:DNA Methylation; D005260:Female; D006367:HeLa Cells; D052162:Human papillomavirus 16 / Q000235:genetics; D006801:Humans; D008745:Methylation; D009856:Oncogene Proteins, Viral / Q000235:genetics* / Q000378:metabolism; D011401:Promoter Regions, Genetic; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D034741:RNA, Small Interfering / Q000008:administration & dosage / Q000235:genetics; D012097:Repressor Proteins / Q000235:genetics* / Q000378:metabolism; D019098:Telomerase / Q000096:biosynthesis / Q000235:genetics* / Q000378:metabolism; D014162:Transfection; D002583:Uterine Cervical Neoplasms / Q000201:enzymology / Q000235:genetics* / Q000378:metabolism / Q000821:virology
chemical_list: C068651:E6 protein, Human papillomavirus type 16; D009856:Oncogene Proteins, Viral; D012333:RNA, Messenger; D034741:RNA, Small Interfering; D012097:Repressor Proteins; C509186:TERT protein, human; D019098:Telomerase