The microstructure evolution and mechanical properties of GH4169 alloy during cold rolling and heat treatment were investigated by scanning electron microscope, electron backscatter diffraction, and transmission electron microscope. Results show that the grains are elongated into fibers with increasing the cold rolling deformation degree, and no δ phase can be observed in the microstructure. After heat treatment, the original deformed grains are replaced by small recrystallized grains, and the grain size is decreased with increasing the cold rolling deformation degree and decreasing the heat temperature. However, the mixed grain structure appears after cold rolling deformation of 50%. When the heat treatment temperature is 950 and 990 °C, the δ phase is precipitated in the matrix. The content of δ phase is increased with increasing the deformation degree and the morphology is changed from short rods into spherical shapes. When the deformation degree is 70%, the ultimate tensile strength (UTS) reaches 1484.27 MPa, which is 1.92 times higher than that of the cold rolled alloy (772.5 MPa). However, the elongation (EL) decreases to 8.93%, and it increases to 46.47% after heat treatment at 990 °C, which is 5.2 times higher than that of the cold-rolled alloy. The optimal combination of mechanical properties (UTS=943.59 MPa, EL=52.31%) can be achieved when the cold rolling deformation degree is 50% and the heat treatment temperature is 990 °C.