Objective: To evaluate the impact of GOLPH3 expression in cumulus granulosa cells on the outcomes of intracytoplasmic sperm injection (ICSI).
Methods: A total of 119 women receiving ICSI due to male infertility at our center between April, 2012 and June, 2014 were enrolled in the study. Cumulus granulosa cells were collected from the women for detection of GOLPH3 expressions using immunocytochemistry, Western blotting, and real-time PCR. GOLPH3 expression rate was compared between women with and without clinical pregnancy following ICSI, and the associations of GOLPH3 expression with the laboratory indicators of ICSI outcomes were assessed.
Results: Immunocytochemistry showed that GOLPH3 expression was located mainly in in the plasma of the cumulus granulosa cells. The rate and intensity of GOLPH3 expression in the cumulus granulosa cells differed significantly between women with and without clinical pregnancy following ICSI (P<0.05). GOLPH3 expression was found to positively correlate with the numbers of punctured follicles, grade III oocyte cumulus complex, ICSI oocytes, fertilized oocytes, cleavage, high quality embryos, blastocysts, high quality blastocysts, and frozen embryos (all P<0.01). The results of RTPCR and Western blotting revealed significant differences in GOLPH3 expressions at both the mRNA and protein levels in the cumulus granulosa cells between the pregnant and non-pregnant groups after ICSI (t=14.560, P=0.000). Western blot analysis revealed significant difference of GOLPH3 protein expression in cumulus granulosa cells between women with and without clinical pregnancy following ICSI.
Conclusion: GOLPH3 expression in the cumulus granulosa cells plays an important role in the development of oocytes and promotion of conception to affect the outcomes of ICSI.
目的: 探讨卵丘颗粒细胞高尔基磷酸化蛋白-3（GOLPH3）表达对单精子卵胞浆内注射（ICSI）助孕结局的影响。
方法: 收集2012年4月~2014年6月，因男性不育症行ICSI助孕的119例女性患者的卵丘颗粒细胞，采用细胞免疫化学，Western bloting方法检测颗粒细胞GOLPH3蛋白的表达，采用real-time荧光定量PCR方法检测GOLPH3基因表达。比较分析ICSI妊娠组与非妊娠组病例卵丘颗粒细胞GOLPH3表达率差异，并分析它们与ICSI实验室结局各项指标：穿卵泡数、获卵数、获卵率；受精数、受精率；卵裂数、卵裂率；优质胚胎数、囊胚形成数及囊胚冷冻数之间的相关关系。
结果: 对119份卵丘颗粒细胞涂片进行免疫细胞化学分析GOLPH3的表达，结果发现GOLPH3主要表达在细胞质中，妊娠组与未妊娠组卵丘颗粒细胞GOLPH3表达率与表达强度组间差异均有统计学意义（P均 < 0.05）。卵丘颗粒细胞GOLPH3表达率与ICSI实验室结局指标：穿卵泡数，Ⅲ级OCCs数，ICSI卵子数，受精数，卵裂数，平均优质胚胎数，平均囊胚数，优质囊胚数，平均冷冻胚胎数等呈显著正相关关系（P均 < 0.01）。Real-Time PCR，Western blot检测妊娠组与未妊娠组颗粒细胞GOLPH3蛋白与基因表达组间差异也均有统计学意义（P均 < 0.05）。
结论: GOLPH3在卵丘颗粒细胞的表达，可能影响卵母细胞的成熟，卵子的发育能力，进而与ICSI助孕结局有关。