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Published on: April 10, 2018
LncRNA MALAT1 was regulated by HPV16 E7 independently of pRB in cervical cancer cells
Ting Wang1, Wei Zhang1, Wenbin Huang2
1Key Laboratory of Developmental Genes and Human Disease in Ministry of Education, Department of Biochemistry and Molecular Biology, Medical School of Southeast University, Nanjing, 210009, China.
Abstract:
High-risk human papillomavirus (HPV) infection was one of the first step in the process of carcinogenesis in cervical cancers. The expression of viral oncoprotein E7 was essential in this process by inactivating the tumor suppressor proteins RB, in addition to interacting with other host proteins. LncRNA MALAT1 was found to be altered in human cervical cancer tissues, suggesting an important role in tumorigenesis. Moreover, MALAT1 was also overexpressed in HPV16 positive cervical cancer cell lines in an HPV16 E7 dependent manner. To explore the mechanism of E7 involved in MALAT1 up-regulation, the deletion mutant E7∆N and E7∆C were constructed to test the functional domain of E7 for MALAT1 regulation. ChIP, EMSA and UV crosslink were performed to detect the interaction between E7 and MALAT1 promoter. E7 and E7∆N mutant were observed that could bind with MALAT1 promoter directly and interacted with SP1 to form triple complex. E7-SP1 interaction contributed to the transcription activation of MALAT1 promoter. E7 and E7∆N also could promote cell proliferation, invasion, and migration, and the stimulating effect could be reversed by siMALAT1. Here we showed that HPV16 E7 as well as E7∆N could associate with SP1 and bound directly to MALAT1 promoter in vitro and in vivo. This function way of E7 was independent of pRB in cervical cancer cells. To our knowledge, this was the first reported function model for E7 as transcription activator to directly bind to the target promoter.
Insights
Human papillomavirus (HPV) oncoprotein E7 directly activates long non-coding RNA MALAT1 transcription by binding to its promoter, promoting cervical cancer cell growth. This mechanism is independent of pRB, offering new therapeutic targets.
Area of Science:
- Molecular Oncology
- Viral Carcinogenesis
Background:
- High-risk human papillomavirus (HPV) infection is a key step in cervical cancer development.
- HPV oncoprotein E7 inactivates tumor suppressor protein RB and interacts with host proteins.
- Long non-coding RNA MALAT1 is dysregulated in cervical cancer and overexpressed in HPV16-positive cell lines.
Purpose of the Study:
- To elucidate the mechanism by which HPV oncoprotein E7 up-regulates MALAT1.
- To investigate the functional domains of E7 involved in MALAT1 regulation.
- To explore the interaction between E7, MALAT1 promoter, and host factors like SP1.
Main Methods:
- Construction of E7 deletion mutants (E7∆N, E7∆C).
- Chromatin immunoprecipitation (ChIP), Electrophoretic Mobility Shift Assay (EMSA), and UV crosslinking assays.
- In vitro and in vivo studies to assess E7-promoter interactions and effects on cell proliferation, invasion, and migration.
Main Results:
- HPV16 E7 and E7∆N mutant directly bind to the MALAT1 promoter.
- E7 interacts with SP1, forming a triple complex that activates MALAT1 transcription.
- E7-induced cell proliferation, invasion, and migration are reversed by MALAT1 knockdown.
Conclusions:
- HPV16 E7 acts as a transcription activator by directly binding to the MALAT1 promoter, independent of pRB.
- The E7-SP1 interaction is crucial for MALAT1 promoter activation.
- This study reveals a novel mechanism of E7-mediated oncogenesis via MALAT1 upregulation.
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