Background: Chemotherapy knowledge is a term that generally describes the information patients receive about treatment goals, duration, possible side effects and lifestyle adjustments. Knowledge and attitude towards cancer treatment have been found to significantly impact help seeking in patients’ adherence to treatment. Objective: To assess knowledge and attitude of patients on chemotherapy and associated factors at oncology center of Amhara Regional State, North West Ethiopia, 2023. Methods: An institution-based cross-sectional study design was conducted on 423 systematically selected cancer patients who receive chemotherapy by using structured interviewer administered questionnaires and chart reviews. Data was entered into Epi-data version 4.6 Software and exported to statistical package for social science version 26 for analysis. Bivariable and multivariable analysis using a logistic regression model were used to determine the association between predictor variables and the dependent variable. Model of fitness was checked by using Hosmer and Lemeshow test. Variables those have a p-value of <0.25 in Bivariable logistic regression were entered in to multivariable logistic regression’s-values<0.05 were considered significantly associated to outcome variables. Results; the totals of 405 Participants were participated in study. About 46.2%, 51.4% participants had good knowledge and favorable attitude towards chemotherapy respectively. For knowledge uneducated (AOR=0.5,95%CI:0.31,0.83), stage-II cancer (AOR=0.53,95%CI:0.31,0.89),stage-III(AOR=0.39,95%CI:0.21,0.70)and age36- 55(AOR=1.81,95%CI:1.13,2.91).for attitude, uneducated(AOR=0.57,95%CI:0.37,0.87) ,family history of cancer(AOR=2.1,95%CI:1.87,3.68)and community health insurance have 2.5(AOR=2.5: 95%CI: 1.474-4.083) were significantly associated . Conclusions and Recommendations; About less than half of cancer patients had good knowledge and half of participants had favorable attitude about cancer therapy and predictors such as age, education and stage of cancer were found to be significantly associated with good knowledge. Education, member of health insurance, family history of cancer wear significantly associated with favorable attitude. There for expand community health insurance and establish a health information delivery system. Keyword; chemotherapy, attitude, knowledge, Ethiopia