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A selective role for CDCA7 in lymphomagenesis
of lymphoma cells. However, high CDCA7 levels in immortal, but non-malignant B cells, are not sufficient to promote anchorage-independent growth because forced expression of CDCA7-1 or CDCA7-2 in non-transformed JY cells or in JY cells transduced with lentivirus encoding constitutively active H-RAS (R12,T59) did not induce their growth in soft agar (Online Supplementary Figure S9). These results support the notion that malignant transfor- mation requires the acquisition of multiple genetic lesions beyond those necessary to proliferate indefinitely.
Figure 6. CDCA7 mediates BL tumor formation. DG-75, BL2 and Ramos cells were transduced with lentivirus encoding sh-Ctl, sh-25 or sh-83 and selected in the presence of puromycin >5 days. A. DG75, B. Ramos, and C. BL2 cells trans- duced with lentiviruses encoding the indicated shRNAs were inoculated subcu- taneously in immunodeficient NOD-SCID mice. Tumors were extracted after 3 weeks. Circles, squares and triangles indicate the weight of individual tumors and horizontal bars indicate the mean (long bar) and s.e.m (short bar). *P<0.05, **P<0.01 and ***P<0.001 vs. sh-Ctl; one-way ANOVA with Bonferroni post-test.
CDCA7 mediates BL lymphoma growth in vivo
We have shown that DG-75, BL2, and Ramos BL cells are tumorigenic in immunodeficient mice, while EBV-immor- talized B-cells are not.30,32 To investigate the contribution of CDCA7 to the in vivo tumorigenicity of BL cells, we trans- duced DG-75, BL2, and Ramos BL cells with sh-Ctl, sh-25 or sh-83 and subcutaneously inoculated them in immun- odeficient NOD-SCID mice. As expected, control-trans- duced DG-75, Ramos and BL cells readily elicited growth of large tumors in these mice (Figure 6). Importantly,
A
B
C
Figure 7. CDCA7 is overexpressed in various types of lymphoid malignancies.
Representative immunoblot analysis of CDCA7 expression in the indicated BL, LCL, Diffuse Large B-Cell lymphoma (DLBCLs), Follicular Lymphoma (FL), Mantle Cell Lymphoma (MCL), uncharacterized non-Hodgkin Lymphoma (N-HL), Acute Lymphocytic B-cell leukemia (B-ALL) and T-cell leukemia (T-ALL), and Myeloid leukemia (ML) cell lines. Tubulin is shown as loading control.
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