Chemoresistance is a main cause for the failure of cancer management and intensive investigation is on-going to control chemoresistant (CR) cancers. Although NF-κB has been suggested as one of the potential targets to alleviate chemoresistance of epithelial ovarian cancer (EOC), direct targeting of NF-κB may result in an unexpected effect due to the complex regulatory network via NF-κB. Here we show that AIMP2-DX2, a splicing variant of tumor suppressor AIMP2, can be a therapeutic target to control CR EOC. AIMP2-DX2 was often highly expressed in CR EOC both in vitro and in vivo. AIMP2-DX2 compromised the tumor necrosis factor alpha-dependent pro-apoptotic activity of AIMP2 via the competitive inhibition of AIMP2 binding to TRAF2 that plays a pivotal role in the regulation of NF-κB. The direct delivery of siRNA against AIMP2-DX2 into abdominal metastatic tumors of ovarian cancer using a microneedle converged on microendoscopy significantly suppressed the growth rate of tumors. The treated cancer tissues showed an enhanced apoptosis and the decreased TRAF2 level. Thus, we suggest that the downregulation of AIMP2-DX2 can be a potent adjuvant therapeutic approach for CR EOC that resulted from an aberrant activity of NF-κB.