[Effects of down-regulating Spag6L expression on the proliferation and apoptosis of mouse spermatocytes].

ABSTRACT

OBJECTIVE: To explore the expression of the Spag6L gene during spermatogenesis and the effects of Spag6L silencing on the proliferation and apoptosis of mouse GC-2 spd cells. METHODS: Using reverse-transcription PCR and real-time qPCR, we detected the expression of the Spag6L gene in the testis tissue collected from the mice at 8, 16, 20, 28 and 42 postnatal days. We prepared lentiviral particles inhibiting the expression of Spag6L and transfected them into the GC-2 spd cells. Then we screened the stably transfected cell lines with the Spag6L expression effectively down-regulated by real-time qPCR, analyzed the effects of Spag6L silencing on the proliferation, activity, cell cycle and apoptosis of the GC-2 spd cells by cell counting and flow cytometry, and on the expression levels of pro-apoptotic Bax and anti-apoptotic Bcl-2 by Western blot. RESULTS: The Spag6L gene was slightly expressed in the testis tissue of the mice at 8 postnatal days and gradually up-regulated with the development of the testis. Inhibition of the Spag6Lexpression significantly decreased the activity of the GC-2 spd cells (P < 0.01), leading to cell arrest in the G1 phase. The expression of the Bax protein was dramatically up-regulated (P < 0.01) while that of Bcl-2 remarkably down-regulated (P < 0.01) in the Spag6L shRNA- transfected cells, inducing the apoptosis of the cells. CONCLUSIONS: The Spag6L gene is involved in the spermatogenesis of mice by regulating the cell cycle, proliferation and apoptosis of spermatocytes.