Previous research has revealed that n3 polyunsaturated fatty acids (PUFAs) exhibit anticancer activities. Lipids from a fish brain contain substantial n3 PUFAs. However, no research has been conducted on the action and mechanism of their potent anticancer activities. In this study, total lipids (TLs) from silver carp brain were isolated into polar lipids (PLs) and neutral lipids (NLs), and the anticancer potential of the lipid fractions (LFs) was investigated using the human cervical carcinoma HeLa cell line. LFs effectively inhibited the cell proliferation of HeLa cells in a time- and dose-dependent manner by cell cycle arrest at the S stage and by inducing apoptosis. Further analyses indicated that the loss of mitochondrial membrane potential could be one of mechanisms of apoptosis induced by LFs. Among the TLs, PLs have proven to be more effective in inducing cervical carcinoma cell death than NLs. This work will play a role in promoting lipids from silver carp brain as a potential preventive and therapeutic agent against human cervical carcinoma.