Objective To observe the expression of sperm protein (SP) 32/OY-TES-1 in retinoblastoma (RB).Methods Thirty paraffin specimens of pathologically confirmed RB eyeballs were investigated in this study. The SP32/OY-TES-1 mRNA expression of 15 RB tissues, 12 non-tumor retinal tissues and 22 normal eye tissues were detected by reverse transcription polymerase chain reaction (RT-PCR). The SP32/OY-TES-1 protein expression of 30 RB tissues and 24 normal retinal tissues were examined by immunohistochemistry. The relationship between SP32/OY-TES-1 mRNA, protein expression and age, gender, tumor size, tumor differentiation, clinical stage of RB children were analyzed. Results The expression rate of SP32/OY-TES-1 mRNA in RB tissues, non-tumor retinal tissues and normal eye tissues were 86.7%, 16.7% and 36.4% respectively. Compared the expression rate of SP32/OY-TES-1 mRNA in different gender (chi;2=0.744)，age（chi;2=1.178），clinical stage(chi;2=1.188)，tumor size (chi;2=0.216)，tumor differentiation(chi;2=1.885)，the differences were not statistically significant (P＞0.05). The expression rate of SP32/OY-TES-1 protein in RB tissues was 73.3% and no expression in normal retinal tissues. Compared the expression rate of SP32/OY-TE-1 protein in different gender (chi;2=1.000)，agewith le;two years and ＞two years（chi;2=0.403），tumor size (chi;2=2.274)，tumor differentiation(chi;2=0.138), the differences were not statistically significant (P＞0.05); but in clinical stage (chi;2=6.193)，with or without optic nerve infiltration (chi;2=4.535), the differences were statistically significant(P＜0.05). Conclusions SP32/OY-TES-1 is highly expressed in RB. The expression rate of SP32/OY-TES-1 is related to optic nerve infiltration and clinical stage of RB.