Abstract
Objective
The study was to explore chemokine receptor CXCR3 influence on invasion of T lymphocytic leukemia cell.
Methods
Mouse CXCR3 gene was amplified from T vector containing mouse CXCR3 by PCR, and constructed the mouse CXCR3 overexpressing lentivirus carrying GFP & Puromycin (puro). Jurkat cell line was infected by the constructed CXCR3 overexpression lentivirus. Puromycin was added to screene the successfully infected lymphoblastic leukemia cells. CXCR3 expression on Jurkat cells was analyzed by flow cytometre. Constructed CXCR3-LV5-Jurkat cells were added into the Transwell upper chamber covered with Matrigel. Ligand of CXCR3(CXCL10) was added to the lower chamber. Cell number in the upper chamber and lower chamber which used to calculate the invasion rate was counted by flow cytometry respectively.
Results
GFP+ Jurkat cells was observed less than <10% after lentivirus infection 96h. GFP+Jurkat cells achieved to 90% by Puromycin selection. The expression of CXCR3 on CXCR3-LV5-Jurkat cell group was 90% higher than LV5-Jurkat cell group. CXCR3-LV5-Jurkat cell line stably overexpressed CXCR3 was successfully constructed. The invasion rate of CXCR3-LV5-Jurkat cell lines was significantly higher than LV5-Jurkat control group (12.71 %± 1.03% vs 6.82% ± 0.49%, p <0.0001).
Conclusion
Overexpression of CXCR3 on T lymphocytic leukemia cell line (Jurkat cell) can enhance the invasion of leukemia cells in vitro, CXCR3/CXCL10 path way may induce the T lymphoblastic leukemic cell invading the central nervous system.
No relevant conflicts of interest to declare.
Author notes
Asterisk with author names denotes non-ASH members.
