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Testicular germ cell tumour (TGCT) is the most common cancer in young men in large parts of the world, but the aetiology is mainly unknown. Genome-wide association studies have so far identified about 50 susceptibility loci associated with TGCT, including  SPRY4 . SPRY4 has shown tumour suppressor activity in several cancer cells, such as lung and prostate, while it was found to act as an oncogene in ovarian cancer. An intronic region within the  SPRY4  gene produces a long non-coding RNA,  SPRY4-IT1 , which has been reported to act as an oncogene in melanoma, breast cancer, and colorectal cancer, and as a tumour suppressor in lung cancer. The roles of  SPRY4  and  SPRY4-IT1  in TGCT development are yet unknown. We found higher expression levels of  SPRY4 , both mRNA and protein, and of  SPRY4-IT1  in human TGCT than in normal adult testis. Small-interfering RNA (siRNA)-mediated transient knockdown of  SPRY4  and  SPRY4-IT1  in two TGCT cell lines 833 K and NT2-D1 resulted in decreased cell growth, migration, and invasion. Knockdown of  SPRY4  and  SPRY4-IT1  also led to a significant reduction in the phosphorylation of Akt. Our findings indicate that  SPRY4  and  SPRY4-IT1  may act as oncogenes in TGCTs via activation of the PI3K / Akt signalling pathway.

Knockdown of SPRY4 and SPRY4-IT1 inhibits cell growth and phosphorylation of Akt in human testicular germ cell tumours