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
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