Wearable sensors play a key role in point-of-care testing (POCT) for their flexible and integration capability for sensitive physiological and biochemical sensing. Here, we present a multifunction wearable silk patch with both electronic channels and microchannels by utilizing matrix-assisted sacrificial 3D printing methods. Owing to the unique properties of a composite silk film (polyvinyl alcohol (PVA) and silk fibroin (SF)), the wearable sensors possess excellent tensile properties, self-healing ability and biocompatibility. Multi-layer channel (microfluidics and microcircuit)-integrated silk wearable sensors were then fabricated for simultaneous sensitive sensing of human cancer markers (carcinoembryonic antigen (CEA) and alpha-fetoprotein (AFP)) and motion monitoring. These features of the silk wearable sensors indicate their potential value for sensitive sensing, which will enable them to find broader applications in many fields in POCT, artificial skin and organ-on-a-chip systems.