Abstract: Healthcare professionals of chemotherapy departments are in a regular contact with anticancer drugs that have genotoxic properties. The analysis of the cytogenetic status of healthcare professionals (54 individuals) was for the first time carried out using a new promising noninvasive method, buccal micronucleus cytome assay with the calculation of integral indices. The effect was estimated with different duration of exposure to chemotherapy drugs (after an employee’s vacation, one month after vacation, and before vacation). Statistically significant increases in almost all studied indices of cytogenetic damages, proliferation, and apoptosis as compared with the control group of office staff (47 individuals) were detected. Among healthcare professionals, an increase in the portion of individuals with an increased level of cytogenetic abnormalities with the maximum exposure duration from 24 to 38% (11% in the control) was established. Among cancer patients (9 individuals), the portion of individuals with a high level of cytogenetic abnormalities was 56% before the next course of chemotherapy and 87% after it. © 2022, Pleiades Publishing, Inc.