Electric field measurement techniques based on Rydberg atoms have proven to have unique advantages in high sensitivity and have drawn wide attention in microwave electric field metrology and communication. The Rydberg states employed for microwave sensing are usually excited by two- or three-step laser irradiation. It leads to the Rydberg atoms being populated by some specific magnetic quantum number states determined by the laser polarization configurations, whose further coupling to another Rydberg state is also constrained by the microwave polarization. Based on a three-laser optical excitation, we demonstrate this effect on the microwave spectra of the 85Rb transitions 41F7/2 → 42D5/2 and 41F7/2 → 41G7/2 at various combining polarization configurations. It shows a strong polarization dependence on the microwave spectrum and then affects the sensitivity of Rydberg atom-based microwave field sensing. Advisable polarization schemes for higher sensitivity are suggested by the two types of transitions.