Metallothioneins (MTs) are proteins known to be involved in defense mechanisms against heavy metals and reactive oxygen species. In human, more than ten MT isoform genes have been identified, in contrast to much fewer isoforms in other mammalian species. The increased number of isoforms in human may have some biological significance; for example, isoforms may have been functionally differentiated to deal with various environmental factors in the evolutional process. However, we know little about the functions of the individual MT isoforms. To clarify functional differences between human MT isoforms, we developed a method to determine individual isoform mRNA levels using real-time polymerase chain reaction (PCR), and studied responses of the isoform genes against heavy metals (Zn, Cd, Cu) and As in HeLa cells. These metals induced all MT isoforms except for MT-1A by Cu, though their induced levels were different. Furthermore, these metals preferentially induced isoforms MT-2A and MT-1X suggesting that these isoforms may be important in protecting from their cytotoxicity.