Objective To investigate the regulatory effect and molecular mechanism of solute carrier family 1 member 5 (SLC1A5) -tetraspanin superfamily member 1 (TM4SF1) complex on cisplatin resistance in esophageal squamous cell carcinoma (ESCC) cells. Methods SLC1A5-overexpressing Eca109 cells were constructed using lentiviral vectors, and the effect of SLC1A5 on cisplatin sensitivity was assessed through cell viability assays. Western blotting (WB) was employed to detect SLC1A5 expression in Eca109 cells and cisplatin-resistant Eca109 cells (Eca109-R). SLC1A5 expression was knocked down in Eca109-R cells using lentiviral vectors, and cisplatin sensitivity was examined thereafter. Quantitative real-time polymerase chain reaction (qRT-PCR) was utilized to analyze the influence of SLC1A5 knockdown on the expression of key genes involved in DNA damage repair in Eca109-R cells. In SLC1A5-knockdown Eca109-R cells, cell viability assays were performed to evaluate the sensitivity to cisplatin after RAD50 overexpression. Additionally, Eca109 cells were separately or co-overexpressed with SLC1A5 and TM4SF1 using lentiviral vectors, and the effect of the SLC1A5-TM4SF1complex on RAD50 expression and cisplatin resistance was examined through cell viability assays. Results Compared with control cells, Eca109 cells overexpressing SLC1A5 exhibited enhanced cisplatin resistance (P < 0.05). Eca109-R cells showed increased SLC1A5 protein expression, and knockdown of SLC1A5 cells were more sensitivity to cisplatin (P < 0.05). RAD50 gene expression was significantly downregulated upon SLC1A5 knockdown under cisplatin treatment (P < 0.05). Knockdown of SLC1A5 inhibited RAD50 protein expression, and overexpression of RAD50 in SLC1A5-knockdown cells significantly restored cisplatin resistance (P < 0.05). Co-overexpression of SLC1A5 and TM4SF1 can further up-regulate the expression of RAD50, and the drug resistance of the cells to cisplatin was enhanced compared with the control cells(P < 0.05). Conclusion The SLC1A5-TM4SF1 complex promotes cisplatin resistance in ESCC cells by upregulating RAD50 expression.