Aim - an analysis of levels of steroid hormones in heart cell mitochondria at the stages of B16/F10 melanoma development in combination with chronic neurogenic pain in male and female C57BL/6 mice. The study included male and female C57BL/6 mice (n=336). Experimental groups were: intact animals (♂ n=21; ♀ n=21); controls (♂ n=21; ♀ n=21) with a model of chronic neurogenic pain (CNP); the comparison group (♂ n=63; ♀ n=63) with melanomas (B16/ F10); the main group (♂ n=63; ♀ n=63) (CNP+B16/F10) with melanoma transplanted 3 weeks after the CNP model creation. Levels of estradiol (pg/g protein), estrone (pg/g protein), progesterone (ng/g protein), total testosterone (ng/g protein), and free testosterone (pg/g protein) were determined by ELISA in mitochondrial samples. In animals with CNP, changes in levels of steroid hormones were detected only in heart mitochondria of female mice: estradiol was reduced by 4.6 times, estrone by 2.2 times, progesterone by 1.8 times (p˂0.05), and total testosterone by 2.4 times compared with intact values. Melanoma growth (3 weeks) in females downregulated levels of estradiol by 1.7 times (p˂0.05), estrone by 1.8 times (p˂0.05), progesterone by 2.1 times, and total testosterone by 2.3 times compared with intact values. In males, only total testosterone decreased by 2.1 times during the same period. Melanoma growth in presence of CNP (3 weeks) in females decreased levels of estradiol by 1.6 times (p˂0.05) compared with control val- ues, while males showed reduced levels of estradiol and total and free testosterone by 1.3 times (p˂0.05), 1.9 times (p˂0.05) and 2.7 times, respectively, as well as elevated levels of estrone by 1.6 times (p˂0.05). CNP and the growth of B16/F10 melanoma to a greater extent weaken the heart of females due to the involvement of a wider spectrum of hormones and a decrease in their levels in the heart mitochondria, compared to males. When CNP was combined with B16/F10 melanoma, a more significant hormonal imbalance was determined in males than in females.