Protein profiling and identification of modulators regulated by the E7 oncogene in the C33A cell line by proteomics and genomics.
Human papillomaviruses (HPVs) have been recognized as the primary cause of cervical cancer. HPV 16 E7 binds to tumor suppressor retinoblastoma protein, and interferes with its function, causing release of the transcription factor E2F, which influences expression of cell cycle-related genes. This study was performed to identify the genes and proteins modulated by the HPV E7 oncogene. An HPV-negative cervical cancer cell line (C33A) was prepared to establish a stable cell line expressing E7. In order to analyze the target molecules modulated by E7 expression, we used two approaches: matrix-assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF MS) and DNA microarrays. Forty-seven spots were identified in C33A/E7 by two-dimensional electrophoresis and MALDI/TOF MS. Protein disulfide isomerase A3, integrase interactor 1 protein, growth inhibitory protein, glutathione S-transferase P, and vav proto-oncogene were down-regulated, whereas heat shock 60 kDa protein 1, Ku70 binding protein, alpha enolase, 26S proteasome subunit were up-regulated. A genomic approach using a microarray kit showed that IL-12R beta 1, cytochrome c, and tumor necrosis factor receptor II were induced by the E7 oncogene. These results suggest that E7 can evade immune surveillance by suppressing or inducing these cell signaling factors, cell cycle regulators, and chaperones.
Lee
Kyung-Ae
KA
Laboratory of Cellular Biology, Korea Research Institute of Bioscience and Biotechnology, Yuseong, Daejeon, Korea.
Shim
Jung-Hyun
JH
Kho
Chang Won
CW
Park
Sung Goo
SG
Park
Byoung Chul
BC
Kim
Jae-Wha
JW
Lim
Jong-Seok
JS
Choe
Yong-Kyung
YK
Paik
Sang-Gi
SG
Yoon
Do-Young
DY
D018797:Cell Cycle Proteins / Q000378:metabolism; D045744:Cell Line, Tumor; D016207:Cytokines / Q000378:metabolism; D018076:DNA, Complementary / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000378:metabolism; D050684:E2F Transcription Factors; D015180:Electrophoresis, Gel, Two-Dimensional; D005260:Female; D006801:Humans; D013058:Mass Spectrometry; D020411:Oligonucleotide Array Sequence Analysis; D009856:Oncogene Proteins, Viral / Q000737:chemistry* / Q000378:metabolism; D050725:Papillomavirus E7 Proteins; D040081:Protein Array Analysis; D011506:Proteins / Q000737:chemistry*; D020543:Proteome; D040901:Proteomics / Q000379:methods*; D000090063:Proto-Oncogene Mas; D012333:RNA, Messenger / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D016622:Silver Staining; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D014157:Transcription Factors / Q000378:metabolism; D014162:Transfection; D002583:Uterine Cervical Neoplasms / Q000378:metabolism
D018797:Cell Cycle Proteins; D016207:Cytokines; D018076:DNA, Complementary; D004268:DNA-Binding Proteins; D050684:E2F Transcription Factors; C000715470:MAS1 protein, human; D009856:Oncogene Proteins, Viral; D050725:Papillomavirus E7 Proteins; D011506:Proteins; D020543:Proteome; D000090063:Proto-Oncogene Mas; D012333:RNA, Messenger; D014157:Transcription Factors; C059731:oncogene protein E7, Human papillomavirus type 16