2D nanomaterials are widely investigated for biomedical applications, attributed to their large specific surface area, high therapeutic loading capacity, and unique optical, thermal, and/or electronic characteristics. Lattice defects affect the theranostic performance of 2D nanomaterials significantly by altering their electronic properties and chemical binding. Recent investigations have shown that defect-rich 2D nanomaterials are capable of enhancing tumor treatment through efficient drug delivery, photothermal and photodynamic therapies (PTT and PDT), and improving diagnostics via computed tomography (CT), photoacoustic and magnetic resonance imaging. This review summarizes recent progresses, including synthesis, characterization approach, and applications of defect-engineered 2D nanomaterials that are potentially useful in cancer treatment. The expert opinions are also proposed as the conclusion.