Dataset Information

HPV16 E7 Interactome Proteomics After Anisomelic Acid Treatment

ABSTRACT: Human papillomavirus (HPV) is a major cause of cervical and other anogenital cancers, with the viral oncoprotein E6 playing a central role in carcinogenesis. We previously demonstrated that anisomelic acid treatment reduces E7 protein levels in HPV-positive cancer cells. Here, we employed mass spectrometry-based proteomics to characterize changes in the HPV16 E7 interactome in normal oral keratinocytes following anisomelic acid treatment.

INSTRUMENT(S):

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Cell Culture, Keratinocyte

DISEASE(S): Human Papillomavirus Infectious Disease

SUBMITTER: Michael Santos Silva

LAB HEAD: John Eriksson

PROVIDER: PXD069948 | Pride | 2025-12-10

REPOSITORIES: Pride

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			Action	DRS
	20210730_E7_16_1_AA.raw	Raw
	20210730_E7_16_1_AAMG.raw	Raw
	20210730_E7_16_1_control.raw	Raw
	20210730_E7_16_2_AA.raw	Raw
	20210730_E7_16_2_AAMG.raw	Raw

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Project description:Objective: In past decades, the role of high-risk HPV (HR-HPV) infection in cancer pathogenesis has been extensively studied. The viral E7 protein expressed in pre-malignant cells has been identified as an ideal target for immunological intervention. However, the cultivation of HPV in vitro remains a significant challenge, as well as the lack of methods for expressing the HPV E7 protein and generating replication-competent recombinant viral particles, which posed a major obstacle to further exploration of the function and carcinogenic mechanisms of the E7 oncoprotein. Therefore, it is imperative to investigate novel methodologies to construct replication-competent recombinant viral particles that express the HPV E7 protein to facilitate the study of its function. Methods: We initiated the construction of recombinant viral particles by utilizing the ccdB-Kan forward/reverse screening system in conjunction with the Red/ExoCET recombinant system. We followed the infection of C33A cells with the obtained recombinant virus to enable the continuous expression of HPV16 E7. Afterwards, the total RNA was extracted and performed transcriptome sequencing using RNA-Seq technology to identify differentially expressed genes associated with HPV-induced oncogenicity. Results: We successfully established replicative recombinant viral particles expressing HPV16 E7 stably and continuously. The C33A cells were infected with recombinant viral particles to achieve overexpression of the E7 protein. Subsequently, RNA-Seq analysis was conducted to assess the changes in host cell gene expression. The results revealed an upregulation of the CD36 gene, which is associated with the HPV-induced oncogenic pathways, including PI3K-Akt and p53 signaling pathway. qRT-PCR analysis futher identified that the upregulation of the CD36 gene due to the expression of HPV16 E7. Conclusion: The successful expression of HPV16 E7 in cells demonstrates that the replicated recombinant virus retains the replication and infection abilities of Ad4, while also upregulating the CD36 gene involved in the PI3K-Akt signaling and p53 pathways, thereby promoting cell proliferation. The outcome of this study provides a novel perspective and serves as a solid foundation for further exploration of HPV-related carcinogenesis and the development of replicative HPV recombinant vaccines capable of inducing protective immunity against HPV.

Dataset Information

HPV16 E7 Interactome Proteomics After Anisomelic Acid Treatment

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