Figure 4.
DCBLD2 mediates cisplatin-induced EMT and tumor metastasis. (a,b) A total of 1 × 106 Pc9/cis-luciferase cells transfected with shDCBLD2 or scramble control were inoculated into the right lungs of BALB/c nude mice (n = 7 per group). On day 7, mice were intraperitoneally injected with 4.0 mg/kg cisplatin or an equivalent volume of PBS every 7 days. Thirty-five days after inoculation, mice underwent a bioluminescence test and were sacrificed. The primary tumors in the right lung and the metastatic foci in the left lung were subjected to HE staining or IHC staining. Bioluminescence imaging is shown in (a), and quantitation of bioluminescence signals is shown in (b). (c) Representative IHC images of E-cadherin and Vimentin expression in paraffin-embedded serial sections of the primary lesion (right lung) of orthotopically implanted mice from Figure 3A. Scale bar: 200 μm (left) and 50 μm (right). (d) Western blot assay to confirm the effect of DCBLD2 on EMT in cisplatin treatment. Cells with stable DCBLD2 knockdown or scramble control were cultured in the presence or absence of 2 μg/mL cisplatin for 24 h. (e,f) Dynamic imaging analyses of cell migration in stable DCBLD2 knockdown cells and scramble control cells pretreated with 2 μg/mL cisplatin or vehicle control for 24 h. Cell migration was measured over 16 h by HCS. Representative images recording the single-cell motion track (left panel) and single-cell ultimate displacement (right panel) are shown in (e), and the average speed was measured by Harmony in (f). * p < 0.05, ** p < 0.01, t-test. DDP, refers to cis-diamminedichloridoplatinum (II), also known as cisplatin or cisplatinum.