Cracks often appear on the inner surface of metal thick-walled cylinders with multiple interference fits. Considering that no relatively accurate model exists for the cracks on the interface of multi-layered, rotating, thick-walled cylinders, in this paper, the stress intensity factor is established for a radial penetrating crack on the interface of a multi-layered, rotating, thick-walled cylinder. The parameters included in the equation are the rotation speed, the wall thickness ratio, and the interference. First, finite element software is used to calculate the stress intensity factors of two thick-walled cylinders under an interference fit with a crack on the interface. Then, the equation of the stress intensity factor is fitted with the parameters of contact pressure, crack depth, and wall thickness ratio. Next, the weight function is used to calculate the stress intensity factor for radial penetrating cracks on the inner surface of the cylinder’s interface. Finally, 2D finite element models of the four-layer cylinder with a crack are established to verify the equation.