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Figure 9 | BMC Cancer

Figure 9

From: N-benzyladriamycin-14-valerate (AD 198) exhibits potent anti-tumor activity on TRAF3-deficient mouse B lymphoma and human multiple myeloma

Figure 9

Constitutive c-Myc expression counteracted the effects of AD 198 on the proliferation and survival of human MM cells. The human MM cell line 8226 cells were transduced with a lentiviral expression vector of FLAG tagged c-Myc (pUB-FLAG-c-Myc-Thy1.1, labeled as pUB-FLAG-Myc in the figure). Cells transduced with an empty lentiviral expression vector (pUB-Thy1.1) were used as control in these experiments. (A) Transduction efficiency analyzed by Thy1.1 immunofluorescence staining and flow cytometry. Gated population (Thy1.1+) indicates cells successfully transduced with each lentiviral expression vector. Untransduced 8226 cells were used as negative control. (B) Time-dependent effects of AD 198 on endogenous c-Myc and transduced FLAG-c-Myc protein levels. Transduced 8226 cells (8226/pUB-Thy1.1 or 8226/pUB-FLAG-Myc) were cultured in the absence or presence of 4 μM AD 198. Total cellular lysates were prepared at indicated time points, and were immunoblotted for FLAG, c-Myc, followed by actin. Immunoblots of actin were used as loading control. Results are representative of three independent experiments. (C) Decreased anti-proliferative/survival-inhibitory effects of AD 198 on human MM cells with constitutive c-Myc expression. Transduced 8226 cells (8226/pUB-Thy1.1 or 8226/pUB-FLAG-Myc) were treated with various concentrations of AD 198 for 24 h. Total viable cell numbers were subsequently determined by MTT assay. The graph depicts the results of three independent experiments with duplicate samples in each experiment (mean ± SEM). Similar results were also observed in another human MM cell line LP1 cells.

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