A major problem in treating oesophageal squamous cell carcinoma (ESCC) with cisplatin is the development of drug resistance. In order to determine whether phosphatidylinositol 3-kinase (PI3K)-C2β (encoded by the PIK3C2B gene) reduced the sensitivity of ESCC to cisplatin, transfected Eca109 cells that overexpressed PIK3C2B were produced. Additionally, PI3K-C2β-siRNA was used to silence endogenous PI3K-C2β in EC9706 cisplatin-resistant cells. The relationship between PIK3C2B expression and clinicopathological characteristics was also investigated in samples from 61 patients. The overexpression of PIK3C2B in Eca109 cells significantly inhibited cisplatin-induced apoptosis and cleavage of caspase-3. Knockdown of PI3K-C2β enhanced cisplatin-induced apoptosis in EC9706 cells. PIK3C2B expression was associated with an increased level of phosphorylated Akt. Based on the tumour samples, expression of PIK3C2B was associated with tumour metastasis and in vitro assay suggested that it mediated cell migration. These results indicated that PI3K-C2β, via the Akt signalling pathway, might play a key role in cisplatin resistance and that targeting this pathway might be useful in treating cisplatin-resistant tumours.