Let's say your stripline is on layer 3, and layer 4 is a ground plane.
Case 1: Layer 2 has a copper fill above the line
If layer 2 has an unbroken copper region above the stripline with a low impedance connection to power and/or ground, then it doesn't matter if that layer is a "signal layer" or a "plane layer". There is a ground return path on that layer, and your stripline will behave like a stripline with a reference on that layer.
Case 2: Layer 2 does not have copper fill above the line
If layer 2 does not have an unbroken copper region, (whether it's a "signal layer" or a "plane layer" that happens to have a void in that area) then there is no current return path there, and that layer is not providing the return plane for the stripline.
If layer 1 has an unbroken copper region, then you can calculate the stripline characteristics using the total height from layer 3 to layer 1 for the uppper ground-plane distance.
If neither layer 1 nor layer 2 has an unbroken ground plane above your track, then you don't have a stripline; instead you have a buried microstrip, and you should use a buried microstrip calculator to determine the correct geometry instead of a stripline calculator.
In either of these cases, if layer 2 has tracks running nearby and parallel to your line, there is the possibility of coupling signal from the "stripline" to the other track, which will cause loss on the "stripline" and crosstalk on the other line. Try to route nearby tracks on layer 2 perpendicular to the high speed track on layer 3.