Crossfire will technically work with a second card on a PCIe 2 x4 slot, but it will be an issue in some games. You really want a slot running at PCIe 2.0 x8 at least, or ideally PCIe 3 x8 for crossfire.
Nvidia SLI will ONLY work on slots of x8 or greater, the SLI option doesn't show up if you put the card in a x4 slot. AMD don't impose that same restriction so you can actually enable crossfire... but it's questionable as to whether it's worth it.
I have a motherboard that's limited to x4 on the second slot and spent some time researching the impact I'd have going crossfire. I never found any articles with a definitive answer, but the bits and pieces I found suggested that it will have an impact.