Emerging　evidence　suggests　that　Nanog　is　involved　in　cervical　tumorigenesis.　However,　the　regulating　role　of　Nanog　in　tumorigenesis　and　chemosensitivity　are　still　poorly　understood.　In　this　study,　Nanog　was　disrupted　by　transcription　activator-like　effector　nucleases　(TALEN)　in　Hela　cells　and　its　expression　was　significantly　decreased　in　a　single-cell　derived　sub-clone　with　biallelic　mutations.　The　disruption　of　Nanog　not　only　induced　down　regulation　of　some　other　core　transcription　factor　genes　for　cell　self-renewal,　such　as　Oct4,　Sox2　and　FoxD3,　but　also　led　to　the　down　regulation　of　some　mesenchymal　representative　genes,　vimentin　and　N-adherin,　and　up　regulation　of　the　epithelial　gene,　E-cadherin.　In　addition,　the　invasiveness　and　clonogenicity　of　the　Hela　cells　were　obviously　affected,　and　surprisingly　their　sensitivities　to　anti-cancer　drugs　were　also　significantly　increased　in　vitro.　After　Xenograft　into　nude　mice,　the　growth　volumes　of　the　neoplasms　from　the　Nanog　disrupted　Hela　cells　were　significantly　smaller　compared　with　those　from　wild　type　ones.　In　conclusion,　these　results　suggest　that　disruption　of　Nanog　may　reverse　the　status　of　epithelial-mesenchymal　transition,　which　is　critical　in　tumorigenesis,　and　alleviate　chemoresistance,　as　well　as　their　invasiveness,　in　cervical　cancer　cells.