Cervical cancer remains a frequently-occurring gynecologic health problem posing a great threat to women. Tuftelin1 (TUFT1), an acidic protein possessing secretory capacity, has been reported to drive cisplatin resistance in cervical cancer cells. Accordingly, the current study is intended to figure out the specific impacts of TUFT1 on the aggressive behaviors of cervical cancer cells and make an in-depth study into the related regulatory mechanism. Firstly, analysis of TUFT1 expression in cervical cancer cells was performed by RT-qPCR and Western blotting. Cervical cancer cell proliferation was estimated via CCK-8 and colony formation assays. Wound healing, transwell as well as sphere formation assays were used to appraise cell migration, invasion, and stemness, respectively. Western blotting examined the expressions of metastasis- and stemness-associated factors and RT-qPCR also tested the expressions of stemness-associated factors. Co-IP assay was used to verify the binding between TUFT1 and activating transcription factor 1 (ATF1). Subsequent ATF1 expression was examined by RT-qPCR and Western blotting after TUFT1 was silenced. After co-transfected with TUFT1 interference and ATF1 overexpression plasmids, aforementioned functional experiments were conducted again. Western blotting also analyzed the expressions of epidermal growth factor receptor (EGFR) signaling-associated proteins. The experimental data determined that TUFT1 expression was fortified in cervical cancer cells and TUFT1 absence diminished cervical cancer cell proliferation, migration, invasion, and stemness. Besides, TUFT1 bond to ATF1 and positively modulated ATF1 expression. Moreover, ATF1 elevation countervailed the impacts of TUFT1 insufficiency on the proliferation, migration, invasion, stemness as well as EGFR signaling in cervical cancer cells. Anyway, TUFT1 might interact with ATF1 to elicit pro-proliferation, pro-metastasis, and pro-stemness properties and inactivate EGFR signaling in cervical cancer, supporting that TUFT1 might be valued as a potential hallmark for cervical cancer.