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Objective To analyze the clinical value of combined detection of several serum markers in the treatment and prognosis of patients with alpha-fetoprotein (AFP)-negative hepatocellular carcinoma (HCC) after transcatheter arterial chemoembolization (TACE). Methods A total of 120 patients (from September 2016 to March 2018) diagnosed with HCC followed by TACE are recruited in this study. Serum samples were collected and dynamic markers, serum AFP, carcinoembryonic antigen (CEA), carbohydrate antigen-199 (CA199), CA125, gamma-glutamyltranspeptidase (GGT), interleukin (IL)-2, IL-4, IL-6, IL-10, γ-interferon (IFN-γ), and tumor necrosis factor α (TNF-α) levels were detected in 44 AFP-negative patients and 76 AFP-positive patients prior, 1 and 3 months after TACE. During the follow-up period, the tumor activity of patients was monitored and the clinical value of different marker combinations on the efficacy and prognosis of AFP-negative HCC patients after TACE was analyzed until July 2018. Results There was no statistically significant difference in the pre-operative indicator levels between AFP-negative and AFP-positive patients (P>0.05). The sensitivity, specificity, negative predictive value, and positive predictive value of IL-6 combined with GGT in detecting the efficacy and prognosis of AFP-negative and AFP-positive patients were 95.2%, 87.0%, 87.0%, 95.2% and 85.7%, 59.6%, 70.1%, and 79.1%, respectively. The area under the ROC curve of IL-6 for the prognosis of AFP-negative and AFP-positive HCC patients was 0.839 and 0.666. Conclusion Serum IL-6 combined with GGT can be used as an effective indicator for predicting the efficacy and tumor activity of AFP-negative HCC patients after TACE. DOI: 10.11855/j.issn.0577-7402.2019.05.06