Mucin O-glycosylation plays important roles in many carcinogenic events, and it is initiated by the enzymes: UDP-GalNAc∶polypeptide N-acetylgalactosaminyltransferases (ppGalNAc-Ts). The association of ppGalNAc-T2 expression and Jurkat cells proliferation and migration was investigated. The RNAi and negative control shRNA were synthesized and then inserted into lentivirus vector YH1. After enzyme digestion and sequencing confirmation, each of the recombinant vectors and packaging vectors were cotransducted into 293T cells, and the recombinant lentivirus were packaged. Then Jurkat cells were infected by purified lentivirus, RT-PCR and Western blot showed that ppGalNAc-T2 mRNA and protein expression were remarkably decreased after RNA interference. Furthermore, the proliferation and migration abilities of Jurkat cells were detected. In conclusion, the recombinant lentivirus interfering vector was constructed targeting on ppGalNAc-T2 gene, and decreased expression of ppGalNAc-T2 gene inhibit proliferation and migration of Jurkat cells.