In the current study, we found that the expression levels of key molecules in the TLR4/NO signaling pathway (including TLR4, NF-κB p65, and iNOS) were significantly upregulated in HR-HPV-positive cervical cancer tissues and were also significantly higher in HR-HPV-positive cervical cancer HeLa and CaSki cell lines than in the HPV-negative cervical cancer C33a cell line. This evidence concerns the gene NFKB1 and cervical cancer.