Graphene oxide (GO) was chosen as the Raman enhancement substrate, and the effect of the number of GO layers and probe molecule water soluble copper phthalocyanine (TSCuPc) films thickness on surface enhanced Raman scattering (SERS) was investigated. In our work, spin-coating method was carried out for preparing GO films with different number of GO layers by using different concentrations, and then different TSCuPc films were prepared by adjusting the speed. The results of the experiments showed that: with the increasing of the layer number of GO films, both the polarizability induced by π—π conjugation and local dipole moment induced by oxygen-containing groups increased, resulting in a stronger Raman enhancement presenting a saturated tendency. Meanwhile, the enhancement of characteristic peaks of TSCuPc linearly increased with the increasing of the thickness of TSCuPc films and compared to the influence of GO layer number, the first-layer effect resulting from the thickness of TSCuPc was less pronounced. The TSCuPc molecule prepared by solution method is generally used for hole injection layer of organic optoelectronic devices and there will be of great significance to improve the hole injection efficiency, transmission efficiency and stability of organic optoelectronic devices by researching the surface enhanced Raman spectrum of GO and TSCuPc.