In the present study, we aimed to evaluate the effectiveness of second-line gemcitabine-based chemotherapy regimens on overall survival (OS) in malignant pleural mesothelioma (MPM) patients receiving first-line pemetrexed-based regimens. A total of 73 patients with MPM from Akdeniz University, Acibadem Kayseri Hospital, Kayseri Training and Research Hospital, and Erciyes University were analyzed and evaluated retrospectively as two groups: second-line gemcitabine-based chemotherapy and no second-line chemotherapy. The median OS of patients who received second-line gemcitabine-based chemotherapy was 11.3 (5.1-17.5) months, while it was 9.9 (2.1-17.7) months in the patients who did not receive chemotherapy in the second-line setting (p = 0.056). When we evaluated the survival rate at the 6th, 12th, 18th, 24th, and 36th month, the OS rate of the 36th month was significantly higher in patients who received gemcitabine-based second-line chemotherapy (p = 0.041). When evaluating the OS from diagnosis to death, the median OS values were 20.8 (17.5-24.1) months for first-line pemetrexed-based regimens then second-line gemcitabine alone and 13.1 (9.0-17.1) months supportive care after first-line pemetrexed-based regimens (p = 0.005). According to our results, we may consider gemcitabine-based regimens as second-line chemotherapy after treatment with pemetrexed plus platinum in patients with MPM.