Figure 6

Knockdown of SPINK1 reduces cell intravasation, tumor growth and the lung metastases. (a) Mean weight of the tumor mass collected from pre-fertilized eggs (n=8) implanted with shSCRM and shSPINK1-1 cells. (b) Genomic DNA extracted from the lower CAM was used to measure the intravasated human cells by qPCR using human-specific Alu primers. (c) Upper CAM harvested 3 days post engraftment of the shSPINK1-1 and shSCRM tumor cells, and representative images show the frozen sections stained for hematoxylin and eosin (top panel) and human-specific cytokeratin-18 by immunohistochemistry (bottom panel). (d) Genomic DNA extracted from the lungs and liver of the chick embryo was used to measure the intravasated shSCRM and shSPINK1-1 cells by qPCR using human-specific Alu primers. (e) Mean tumor growth in NOD/SCID mice subcutaneously implanted with shSPINK1-1 or shSCRM cells. Tumor growth was monitored weekly up to 4 weeks. (f) Relative SPINK1 expression measured by qPCR in the xenografts excised from shSPINK1-1 and shSCRM control groups. (g) Genomic DNA extracted from the lung and liver of the tumor-bearing mice was used to measure the intravasated shSCRM and shSPINK1-1 cells by qPCR using human-specific Alu primers. Error bars represent mean±s.e.m. P-values derived from two-sided Student’s t-test, *P<0.05, **P<0.005.