Three extraction methods in terms of organic solvent extraction (OSE), microwave -assisted extraction (MAE) and ultrasonic-assisted extraction (UAE) extraction of anthocyanins from blueberry pomace (BP) were optimized by single factor test and orthogonal test, and anthocyanins components and anti-tumor activity were comparative analysised. The results showed that MAE is superior to OSE and UAE in obtaining anthocyanins content, and the optimum extracting parameters to achieve the highest anthocyanins contents of 46.880.63 mg C3G/g from BP was obtained under the microwave power of 300 W, extraction time of 60 s and solid-to-liquid ratio of 1:40 g/mL. Under the optimal combination of parameters, six anthocyanins components were obtained by the three extraction methods, namely, delphinidin-3-glucoside, cyanidin-3-glucoside, cyanidin-3-rutinoside, petunidin-3-glucoside, malvidin-3-galactoside and peonidin-3,5-two hexoside. The BP extracts obtained by three extraction methods inhibited the growth and the invasion ability of HepG2 hepatoma cells and A549 lung cancer cells, and the inhibition effect of A549 lung cancer cells was stronger.