Protein profiling and identification of modulators regulated by the E7 oncogene in the C33A cell line by proteomics and genomics

Lee, Kyung-Ae; Shim, Jung‐Hyun; Kho, Chang Won; Park, Sung Goo; Park, Byoung Chul; Kim, Jae Wha; Lim, Jong‐Seok; Choe, Yong Kyung; Paik, Sang Gi; Yoon, Do Young

doi:10.1002/pmic.200300626

Cited by 52 publications

(35 citation statements)

References 39 publications

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“…In patients suffering with cervical cancer, Hsp27 is predominantly expressed in well-differentiated SCCs and also moderately differentiated SCCs. Including our results, HSP 27 has recently been shown to be up-regulated in primary carcinoma of the vagina (11), and in C33A cells that were transfected with the HPV E7 gene (17).…”

Section: Discussionsupporting

confidence: 64%

Protein Expression Profile using Two-Dimensional Gel Analysis in Squamous Cervical Cancer Patients

et al. 2006

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Purpose: Screening in cervical cancer is now progressing to discover candidate genes and proteins that may serve as biological markers and that play a role in tumor progression. W e examined the protein expression patterns of the squamous cell carcinoma (SCC) tissues from Korean women with using two-dimensional polyacrylamide gel electrophoresis (2-DE) and matrix assisted laser desorption/ionization-time of flight (MALDI-TOF) mass spectrometer.Materials and Methods: Normal cervix and SCC tissues were solubilized and 2-DE was performed using pH 3～10 linear IPG strips of 17 cm length. The protein expression was evaluated using PDQuest 2-D software TM . The differentially expressed protein spots were identified with a MALDI-TOF mass spectrometer, and the peptide mass spectra identifications were performed using the Mascot program and by searching the Swiss-prot or NCBInr databases.Results: A total of 35 proteins were detected in SCC.17 proteins were up-regulated and 18 proteins weredownregulated. Among the proteins that were identified, 12 proteins (pigment epithelium derived factor, annexin A2 and A5, keratin 19 and 20, heat shock protein 27, smooth muscle protein 22 alpha, α-enolase, squamous cell carcinoma antigen 1 and 2, glutathione S-transferase and apolipoprotein a1) were protein previously known to be involved in tumor, and 21 proteins were newly identified in this study. Conclusion

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Section: Discussionsupporting

confidence: 64%

Protein Expression Profile using Two-Dimensional Gel Analysis in Squamous Cervical Cancer Patients

et al. 2006

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“…Nonetheless, several gene expression profiling studies have been conducted on HPV E7-expressing cells using microarray analysis. The cells employed included mouse cells, cervical cancer cell lines (C33A and CaSki), and PHKs (3,(5)(6)(7)90). However, there are major limitations with the DNA microarray approach, such as the relatively high background noise resulting from solution-based DNA probe hybridization as well as the limited scope of profiling analysis, which is confined by the DNA probe set.…”

Section: Discussionmentioning

confidence: 99%

“…The transforming properties of high-risk HPVs such as HPV 16 (HPV-16) primarily depend on E7 as well as E6 oncogenes (1,2). HPV E6 and E7 proteins promote the degradation of p53 and pRb, respectively (3,4). E7 from the high-risk HPV types can abrogate cell cycle checkpoints and induces genomic instability.…”

mentioning

confidence: 99%

“…E7 from the high-risk HPV types can abrogate cell cycle checkpoints and induces genomic instability. Although several transcription profiling studies for E7 have been conducted using DNA microarray analysis (3,(5)(6)(7), the HPV E7 activities downstream from, or independent of, pRb responsible for deregulation of cell cycle and induction of genomic instability are not fully understood.…”

mentioning

confidence: 99%

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Role of WDHD1 in Human Papillomavirus-Mediated Oncogenesis Identified by Transcriptional Profiling of E7-Expressing Cells

Yang

Zhang

Gao

et al. 2016

J Virol

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The E7 oncoprotein of the high-risk human papillomavirus (HPV) plays a major role in HPV-induced carcinogenesis. E7 abrogates the G 1 cell cycle checkpoint and induces genomic instability, but the mechanism is not fully understood. In this study, we performed RNA sequencing (RNA-seq) to characterize the transcriptional profile of keratinocytes expressing HPV 16 (HPV-16) E7. At the transcriptome level, 236 genes were differentially expressed between E7 and vector control cells. A subset of the differentially expressed genes, most of them novel to E7-expressing cells, was further confirmed by real-time PCR. Of interest, the activities of multiple transcription factors were altered in E7-expressing cells. Through bioinformatics analysis, pathways altered in E7-expressing cells were investigated. The upregulated genes were enriched in cell cycle and DNA replication, as well as in the DNA metabolic process, transcription, DNA damage, DNA repair, and nucleotide metabolism. Specifically, we focused our studies on the gene encoding WDHD1 (WD repeat and high mobility group [HMG]-box DNA-binding protein), one of the genes that was upregulated in E7-expressing cells. WDHD1 is a component of the replisome that regulates DNA replication. Recent studies suggest that WDHD1 may also function as a DNA replication initiation factor as well as a G 1 checkpoint regulator. We found that in E7-expressing cells, the steady-state level of WDHD1 protein was increased along with the half-life. Moreover, downregulation of WDHD1 reduced E7-induced G 1 checkpoint abrogation and rereplication, demonstrating a novel function for WDHD1. These studies shed light on mechanisms by which HPV induces genomic instability and have therapeutic implications. IMPORTANCEThe high-risk HPV types induce cervical cancer and encode an E7 oncoprotein that plays a major role in HPV-induced carcinogenesis. However, the mechanism by which E7 induces carcinogenesis is not fully understood; specific anti-HPV agents are not available. In this study, we performed RNA-seq to characterize transcriptional profiling of keratinocytes expressing HPV-16 E7 and identified more than 200 genes that were differentially expressed between E7 and vector control cells. Through bioinformatics analysis, pathways altered in E7-expressing cells were identified. Significantly, the WDHD1 gene, one of the genes that is upregulated in E7-expressing cells, was found to play an important role in E7-induced G 1 checkpoint abrogation and rereplication. These studies shed light on mechanisms by which HPV induces genomic instability and have therapeutic implications. H uman papillomaviruses (HPVs) are small DNA viruses that replicate in squamous epithelia. Specific types of HPV (highrisk HPVs) are the causative agents for cervical and several other cancers (1). The transforming properties of high-risk HPVs such as HPV 16 (HPV-16) primarily depend on E7 as well as E6 oncogenes (1, 2). HPV E6 and E7 proteins promote the degradation of p53 and pRb, respectively (3, 4). E7 from the high-risk...

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“…In the genomic approach using DNA microarrays the researchers showed that IL-12R , cytochrome c and TNF II were induced by E7 oncogene (K.A. Lee et al, 2004). On the other hand, Yim and coworkers (2004b) analyzed the genomic and proteomic expression patterns in 2 different HPV16 E6 transfected human carcinoma cell lines.…”

Section: Molecular Bases Of Human Papillomavirusmentioning

confidence: 99%

Molecular Bases of Human Papillomavirus Pathogenesis in the Development of Cervical Cancer

Pedroza-Saavedra¹,

Plett-Torres²,

Chihu-Amparán³

et al. 2012

Human Papillomavirus and Related Diseases - From Bench to Bedside - Research Aspects

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Protein profiling and identification of modulators regulated by the E7 oncogene in the C33A cell line by proteomics and genomics

Cited by 52 publications

References 39 publications

Protein Expression Profile using Two-Dimensional Gel Analysis in Squamous Cervical Cancer Patients

Protein Expression Profile using Two-Dimensional Gel Analysis in Squamous Cervical Cancer Patients

Role of WDHD1 in Human Papillomavirus-Mediated Oncogenesis Identified by Transcriptional Profiling of E7-Expressing Cells

Molecular Bases of Human Papillomavirus Pathogenesis in the Development of Cervical Cancer

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