The invention discloses a method for preparing an electrochemical luminescence paper chip and a method for measuring content of signal molecule hydrogen sulfide in MCF-7 by using an electrochemical luminescence sensor. The method is characterized by preparing a hydrophobic region, a semi-hydrophobic region, a hydrophilic region and a hollow channel on paper by utilizing a wax printing technology, cutting the hollow channel by virtue of a laser cutting machine, preparing proper printing ink, printing a corresponding reference electrode and a working electrode on the paper, functionalizing the working area, dripping stock solution in a pretreatment region, and fixing cancer cells; and folding the prepared paper chip, forming a three-electrode system, dropwise adding electrolyte solution in a reaction region, connecting an electrochemical workstation, thus realizing high-sensitivity detection of the low-content signal molecules in the cancer cells.