This study focusses on investigating the effect of flow orientation on global and local two-phase flow parameters under two-phase flow conditions. Flow visualization, pressure measurement, and conductivity probe measurements are performed in a test facility made of 50.8 mm inner diameter acrylic pipes with flow visualization, pressure measurement, and conductivity probe measurement. Characteristics of flow regimes and interfacial structures, and frictional pressure drop prediction in the vertical upward and vertical downward two-phase flows are compared and analyzed. The results show that unique coring phenomenon and slug bubbles with off-centered noses appear in the vertical downward flow. The flow regime transition boundaries shift to the lower gas superficial velocities in the vertical downward flow compared to that in the vertical upward flow. Furthermore, the distribution and one-dimensional transport of the void fraction, interfacial area concentration, bubble velocity, bubble Sauter-mean diameter, and bubble frequency are acquired and compared to study the effect of flow orientation on interfacial structures. The interfacial structure and its development are found to be mainly affected by the lift force and the turbulence related bubble interactions. The frictional pressure drop is acquired and modeled by the Lockhart–Martinelli correlation. The frictional pressure drop is found to be larger in the vertical upward flow than that in the vertical downward flow. The recommended C value for both vertical upward and vertical downward flows are proposed.