Cone beam digital tomosynthesis (CBDT) is a new imaging technique proposed recently as a rapid approach for creating tomographic images of a patient in the radiotherapy treatment room. The purpose of this work is to investigate the feasibility of performing megavoltage (MV) CBDT clinically. A clinical investigational MV-CBDT system was installed on an existing LINAC. After the installation, the treatment machine can be operated in two distinct modes: (1) normal clinical treatment mode; (2) CBDT mode, in which tomographic images of the patient can be obtained using MV-CBDT. Various calibration and phantom measurements were performed on the system, followed by a patient study. Our phantom measurements have shown that: (1) for the same imaging dose, MV-CBDT has the same signal-difference-to-noise ratio as megavoltage cone beam computed tomography (MV-CBCT); (2) MV-CBDT has a better spatial resolution than MV-CBCT in the planes of reconstruction but a worse spatial resolution in the direction perpendicular to the planes of reconstruction. MV-CBDT patient images were also obtained and compared to that of MV-CBCT. We have demonstrated that it is clinically feasible to perform MV-CBDT in the treatment room for image-guided radiotherapy.