Tumor　resistance　to　chemotherapy　is　the　major　obstacle　to　employ　cisplatin,　one　of　the　broadly　used　chemotherapeutic　drugs,　for　effective　treatment　of　various　tumors　in　the　clinic.　Most　acknowledged　mechanisms　of　cancer　resistance　to　cisplatin　focus　on　increased　nuclear　DNA　repair　or　detoxicity　of　cisplatin.　We　previously　demonstrated　that　there　was　a　unique　metabolic　profile　in　cisplatin-resistant　(CP-r)　human　epidermoid　adenocarcinoma　KB-CP　20　and　hepatoma　BEL　7404-CP　20　cancer　cells.　In　this　study,　we　further　defined　hyperpolarized　mitochondrial　membrane　potentials　(δψ　m)　in　CP-r　KB-CP　20　and　BEL　7404-CP　20　cells　compared　to　the　cisplatin-sensitive　(CP-s)　KB-3-1　and　BEL　7404　cells.　Based　on　the　mitochondrial　dysfunction,　mitaplatin　was　designed　with　two　mitochondrial-targeting　moieties　[dichloroacetate　(DCA)　units]　to　the　axial　positions　of　a　six-coordinate　Pt(IV)　center　to　sensitize　cisplatin　resistance.　It　was　found　that　mitaplatin　induced　more　apoptosis　in　CP-r　KB-CP　20　and　BEL　7404-CP　20　cells　than　that　of　cisplatin,　DCA　and　cisplatin/DCA　compared　on　an　equal　molar　basis.　There　was　more　platinum　accumulation　in　mitaplatin-treated　CP-r　cells　due　to　enhanced　transmembrane　permeability　of　lipophilicity,　and　mitaplatin　also　showed　special　targeting　to　mitochondria.　Moreover,　in　the　case　of　treatment　with　mitaplatin,　the　dramatic　collapse　of　δψ　m　was　shown　in　a　dose-dependent　manner,　which　was　confirmed　by　FACS　and　confocal　microscopic　measurements.　Reduced　glucose　utilization　of　CP-r　cells　was　detected　with　specifically　inhibited　phosphorylation　of　pyruvate　dehydrogenase　(PDH)　at　Ser-232,　Ser-293,　and　Ser-300　of　the　E1α　subunit　when　treated　with　mitaplatin,　which　was　indicated　to　modulate　the　abnormal　glycolysis　of　resistant　cells.　The　present　study　suggested　novel　mitochondrial　mechanism　of　mitaplatin　circumventing　cisplatin　resistance　toward　CP-r　cells　as　a　carrier　across　membrane　to　produce　CP-like　cytotoxicity　and　DCA-like　mitochondria-dependent　apoptosis.　Therefore,　mitochondria　targeting　compounds　would　be　more　vulnerable　and　selective　to　overcome　cisplatin　resistance　due　to　the　unique　metabolic　properties　of　CP-r　cancer　cells.　©　2012　American　Chemical　Society.