Cervical squamous intraepithelial lesions (SIL) and cervical cancer (CxCa) develop because of high-risk human papillomavirus (HR-HPV) infection; however, other cofactors such as endogenous hormones could contribute to the development of this neoplasia. This work aims to determine the relationship between serum concentrations of sex steroid hormones, cortisol, prolactin, and insulin and the degree of cervical squamous intraepithelial lesions in Cuban women. A descriptive and cross-sectional study was carried out at the National Institute of Oncology and Radiobiology. It included 205 women who attended a Gynecology consultation for a cytological diagnosis confirmation of SIL during 2018-2020. Blood samples were collected to determine serum concentrations of estradiol, testosterone, prolactin, cortisol, and insulin. A questionnaire about socio-demographic variables, toxic habits, and gynecological and obstetric history of the patients was applied. Were found high-grade SILs mainly in women between 36 and 45-years-old and those who had one or two births. Serum concentrations of testosterone and insulin were the only ones that showed significant differences between women who presented SIL concerning negative cytology. A 3,23-fold (CI 95% 1.55-6.73) risk of developing SIL was found when testosterone concentrations were superior to 2 nmol/L and a 2,63-fold (CI 95% 1.36-5.09) risk when the insulin concentration was greater than 12 μU/mL. The association of elevated serum concentrations of testosterone and insulin with the presence of SIL may be related to increased bioavailability of estrogens, which may cooperate with viral oncogenes in cervical carcinogenesis. Additionally, mitogenic activity of insulin and its influence on increased IGF-1 bioactivity may facilitate malignant transformation and viral survival. New research is required to determine the mechanisms involved in this process and elucidate whether there is a cooperation between both hormones during the progression to CxCa.