Background With the advancement of cervical cancer elimination strategies, promoting human papillomavirus (HPV) vaccination is essential to achieving this goal. The issue of how to structure and develop message content to promote HPV vaccination is a debatable issue. Objective The efficacy of gain-loss framing in vaccination contexts is disputed. Our study aimed to elucidate the consequences of message framing on attitudes, intentions, and behavioral tendencies toward HPV vaccination, with the objective of refining message framing strategies and their elements. Methods This systematic review adhered strictly to the PRISMA (Preferred Reporting Items for Systematic Reviews and Meta-Analysis) guideline reporting standards to comprehensively retrieve, extract, and integrate data. We searched databases, including PubMed, Embase, Scopus, and Web of Science, for literature published from database construction to August 15, 2023. Literature screening, data extraction, and quality evaluation were performed by 2 researchers. Intervention studies published in English, conducted with populations with children eligible for HPV vaccination, and involving message framing were included. Attitudes, intentions, and behaviors served as outcome evaluation criteria. Results A total of 19 intervention studies were included. Gain-loss framing had no clear effect on vaccination attitudes nor intentions. Loss framing showed a weak advantage at improving HPV vaccination attitudes or intentions, but the evidence was not strong enough to draw definitive conclusions. The impact of gain-loss framing on HPV vaccination behaviors could not be determined due to the limited number of studies and the qualitative nature of the analysis. Conclusions Combining gain-loss framing with other message framing approaches may be an effective way to enhance the effect of message framing. More high-quality message framing content and exploring alternative moderator or mediator variables are required to support the conclusion. Trial Registration CRD42023451612; https://www.crd.york.ac.uk/PROSPERO/display_record.php?RecordID=451612