Figure 4.

Combining Obatoclax with cisplatin and/or pre-miR-34a can impact expression levels of Bcl-xL and Bcl-2 as well as expression levels of cyclin D1 and cdk4/6. In TCCSuP cells, the Obatoclax + cisplatin combination treatment was the most effective treatment with regards to its ability to decrease the expression levels of Bcl-xl and Bcl-2. In 5637 cells, this combination treatment caused a similar impact on these molecules compared to treatment with Obatoclax as a single agent. Combining pre-miR-34a with Obatoclax and/or cisplatin caused increased levels of Bcl-2 expression both TCCSuP and 5637 cells, and increased levels of Bcl-xL in 5637 cells compared to treatment with Obatoclax and/or cisplatin. Treatment with Obatoclax as a single agent or in combination with cisplatin reduced the expression of cyclin D1 and Cdk6 in both cell lines. Treatment with pre-miR-34a also caused the reduced expression of cyclin D1 and Cdk6; however, it caused increased Bcl-2 expression and negated the effect on Obatoclax on Bcl-2 expression in the combination treatments in both cell lines. Unlike the other treatments, pre-miR-34a was also able to reduce Cdk4 expression levels at the treatment doses used for this experiment (Obatoclax; 25, 60, and 40 nM, respectively, in T24, TCCSuP, and 5637, cisplatin; 5 μM, pre-miR-34a; 50 nm). (Veh = vehicle control, Cis = cisplatin, 34a = pre-miR-34a, Obato = Obatoclax, triple combo = Obatoclax + cisplatin + pre-miR-34a).