ABSTRACT: Objective To establish and authenticate an oxaliplatin(L-OHP)resistant human colon cancer cell line,and to explore the mechanism of L-OHP resistance. Methods Resistance to oxaliplatin was induced in HCT-8(HCT-8/L-OHP)colorectal adenocarcinoma cell lines by exposing cells to increasing concentrations of L-OHP. MTT, Flow cytometry(FCM), RT-qPCR, and Western-blot were used to evaluate the resistance to L-OHP. Results Built the stable HCT-8/L-OHP cell model. The drug resistance index(RI)of HCT-8/L-OHP cell line to the L-OHP, 5-FU, CPT-11, VCR, and CDDP were 10.87, 5.35, 3.23, 1.28, and 6.64, respectively. Increased expression of multi-drug resistant genes ABCC1, ABCB1, GSTP1, and ERCC1 were detected in HCT-8/L-OHP cell line. Conclusion We successfully established the L-OHP resistant HCT-8/L-OHP cell line. In addition, the elevated expression of ABCC1, ABCB1 and ERCC1 genes may be associated with molecular mechanism in oxaliplatin resistance. And HOXB8 overexpression is involved in L-OHP resistance mechanism of human colon cancer.