Background: First-line chemotherapy (CT1) with the combination of gemcitabine (gem) + platinum has become a new standard in advanced biliary tract cancer (ABTC) but data on second-line CT (CT2) are lacking. The aim of this study was to evaluate the efficacy and tolerability of CT2 in patients (pts) with ABTC who received gem-platinum in CT1. Methods: We retrospectively reviewed data of consecutive patients who received CT2 for ABTC after failure to gem-platinum in 17 French institutions from November 2002 to December 2013. Progression-free survival (PFS) and overall survival (OS) were estimated from the start of L2 CT using Kaplan Meier method. Cox models were applied for multivariate analyses. Results: Among 603 pts who were treated by gem-platinum in CT1, 196 pts (median age, 63 years, range: 28-82; male, 51.5 %) received a CT2. CT1 included gem + cisplatin (7%) or oxaliplatin (ox) (93%), with a median PFS of 9.7 months (mo) and an ORR of 31%. Characteristics at the beginning of CT2 were: metastatic disease, 94% ; 1-2 metastatic sites, 68%; ECOG PS 0-1, 68%. CT2 CT was 5FU-irinotecan (iri) (n=62), 5FU-ox (n=17), 5FU-cisplatin (n=37), 5FU/capecitabine (CAP) (n=39) or other various regimens (n=41). Among the 186 evaluable pts, there were 22 PR (12%) and 70 SD (38%). After a median follow-up of 26.4 months, median PFS and OS were 3.2 and 6.7 mo respectively. There was no significant difference between CT regimens in terms of PFS (5FU-iri, 2.6 mo; 5FU-ox/5FU-cisplatine, 4.0 mo; 5FU/CAP, 3.2 mo and others, 3.7 mo; p=0,27) and OS (6.0 mo, 6.3 mo, 5.6 mo and 9.7 mo respectively; p=0.27) In multivariate analysis, PS 2-3, bilirubine > 17 µmol/L and CA19.9 > 400 UI/mL were significantly associated with a shorter PFS while PS 2-3, CA19.9 > 400 UI/mL and non-response to CT1 with a shorter OS. A grade 3-4 toxicity was observed in 32% of pts (neutropenia, 33%; diarrhea, 17%) and a toxic death occurred in 1.4% of pts. Conclusions: CT2 is associated with a disease control in a half of pts with ABTC who received gem-platinum in CT1. Nevertheless, the short median PFS observed in this study should encourage the evaluation of new treatments in pts with good clinical conditions and an adequate biliary drainage.