Abstract:

Objective To construct the chronic myeloid leukemia （CML） K562 cell line with overexpression of human tongue cancer drug resistance related gene （TCRP1） by lentivirus vector and to test its biological function. Methods The recombinant plasmid of TCRP1 and the packaging plasmid were co-transfected into 293T cells， and the K562 cells were infected after the titer of the virus was determined. Puromycin was used to screen TCRP1 overexpression cell line K562/TCRP1， and the expression of TCRP1 was detected by fluorescence quantitative PCR and Western blot. The proliferation of K562/TCRP1 and its control cell line was detected by continuous cell counting method and CCK-8 method respectively. The drug sensitivity of the two cell lines to different concentrations of imatinib （IM） was analyzed. Results The TCRP1 lentivirus expression vector was successfully transfected into K562 cells. The results of fluorescence quantitative PCR and Western blot showed that the expression of TCRP1 in K562/TCRP1 cell line was significantly higher than that in the control group at the level of mRNA and protein. The results of continuous cell counting and CCK-8 methods showed that the proliferation of K562/TCRP1 cells were enhanced， and the IC50 value of IM-treated K562/TCRP1 cells was significantly higher than that of the control cells （P < 0.05）. Conclusion The K562 cell line with overexpression of TCRP1 was successfully constructed using lentivirus vector， and we found that the overexpression of TCRP1 might enhance the proliferation and IM resistance of K562 cells， which provided a basis for further exploring the possible role of TCRP1 in the pathogenesis of chronic myeloid leukemia.

Key words: tongue cancer drug resistance related gene 1, overexpression, K562 cell line, chronic myelocytic leukemia