The study investigates the mechanism of HPV oncogene expression in cervical cancer, focusing on the integration of HPV DNA into human chromosomes. Despite the presence of multiple integrated HPV DNA copies, the research finds that virus-host fusion transcripts are primarily derived from a single integrated HPV DNA site in HPV16 and HPV18 cervical cancers and cell lines. The host genomic elements near the integrated HPV DNA are crucial for the efficient expression of the viral oncogenes, leading to clonal cell expansion. The fusion RNAs use a host RNA polyadenylation signal downstream of the integration site and involve splicing to host sequences. In cell culture, siRNAs targeting the host portion of the virus-host fusion transcripts effectively silenced viral E6 and E7 expression, inhibited cell growth, and promoted cell senescence in HPV16+ CaSki and HPV18+ HeLa cells. This finding sheds new light on the mechanisms of HPV-induced carcinogenesis and suggests potential therapeutic strategies for precision medicine in HPV-related malignancies.The study investigates the mechanism of HPV oncogene expression in cervical cancer, focusing on the integration of HPV DNA into human chromosomes. Despite the presence of multiple integrated HPV DNA copies, the research finds that virus-host fusion transcripts are primarily derived from a single integrated HPV DNA site in HPV16 and HPV18 cervical cancers and cell lines. The host genomic elements near the integrated HPV DNA are crucial for the efficient expression of the viral oncogenes, leading to clonal cell expansion. The fusion RNAs use a host RNA polyadenylation signal downstream of the integration site and involve splicing to host sequences. In cell culture, siRNAs targeting the host portion of the virus-host fusion transcripts effectively silenced viral E6 and E7 expression, inhibited cell growth, and promoted cell senescence in HPV16+ CaSki and HPV18+ HeLa cells. This finding sheds new light on the mechanisms of HPV-induced carcinogenesis and suggests potential therapeutic strategies for precision medicine in HPV-related malignancies.