Hypoxia is a critical event in solid tumor development, invasion, and metastasis. Cellular adaptation to hypoxic microenvironment is essential for tumor progression and is largely mediated by hypoxia‐inducible factor‐1α (HIF‐1α) through coordinated regulation of hypoxia‐responsive genes. In this study, we found that membrane type‐2 matrix metalloproteinase (MT2‐MMP), one of the matrix metalloproteinase (MMP) family members, was a novel hypoxia‐responsive gene and was upregulated by HIF‐1α under hypoxia. When cancer cells were subjected to hypoxia (1% O₂) treatment, the mRNA and protein levels of MT2‐MMP were significantly increased in a time‐dependent manner in all three tested cancer cell lines including pancreatic cancer cells (PANC‐1), nonsmall cell lung cancer cells (A‐549), and cervix cancer cells (HeLa). Further analyses indicated that there were two hypoxia‐responsive elements (HREs) in the MT2‐MMP promoter, and HRE1 but not HRE2 was essential for MT2‐MMP transcriptional activation under hypoxia. HIF‐1α specifically and directly bound to MT2‐MMP promoter was analyzed by HIF‐1α binding/competition and chromatin immunoprecipitation (ChIP) assays. Furthermore, we found that upregulation of MT2‐MMP under hypoxia could confer resistance to hypoxia‐induced apoptosis and increase invasiveness of cancer cells. These findings provided a new insight into how cancer cells overcome hypoxic stress and trend to survive and invade, demonstrated a new regulatory mechanism of MT2‐MMP expression in caner cells, and also revealed that MT2‐MMP was a novel hypoxia‐responsive gene and was upregulated by HIF‐1α under hypoxia. © 2011 Wiley‐Liss, Inc.