Piperine (PP), an alkaloid from black and long peppers (Piper nigrum Linn & Piper longum Linn), exhibits antitumor activities in vitro and in vivo. We investigated the ability of piperine (PP) to reverse the drug resistance of human cervical cancer cells. In our study, the cervica cancer cells resistant to mitomycin-C (MMC) treatment were used. We found the growth inhibitory effects of piperine on human cervical cancer cell, which were resistant to MMC. Piperine and MMC co-treatment resulted in a dose-dependent suppression of the cell proliferation. Decreasing of phosphorylated-signal transducer and activator of transcription (p-STAT3) was linked to the suppression of p65 by PP and MMC combinational treatment. Additionally, the presence of PP potentiated the effects of MMC on apoptosis induction in cervical cancer cells with drug resistance, which was dependent on Bcl-2 inhibition. The pro-apoptotic proteins of Bax and Bid were up-regulated, accompanied with Caspase cleavage. Moreover, in mice xenograft models, the combined therapy inhibited tumor growth compared to the PP or MMC mono-therapy group. Our data indicated a novel therapeutic strategy of PP to potentiate MMC-induced anti-tumor effect on cervical cancer cells with drug resistance through blocking p-STAT3/p65 and Bcl-2 activation.