Institute of Animal Physiology and Genetics, Academy of Sciences of the Czech Republic, Libechov, Czech Republic.
Fertil Steril. 2013 May;99(6):1739-48. doi: 10.1016/j.fertnstert.2012.12.040. Epub 2013 Jan 30.
To determine whether inhibition of epidermal growth factor (EGF) receptor tyrosine kinase with lapatinib affects oocyte maturation, expression of the cumulus expansion-associated genes such as tumor necrosis factor alpha-induced protein 6 (TNFAIP6) and prostaglandin-endoperoxide synthase 2 (PTGS2), and synthesis of hyaluronan (HA) and progesterone (P) by porcine oocyte cumulus complexes (OCC).
Our work focuses on lapatinib, an orally active small molecule that selectively inhibits the tyrosine kinase domain of both EGF receptor and human EGF receptor 2, and downstream signaling.
A reproductive biology laboratory.
PATIENT(S): Not applicable.
INTERVENTION(S): Porcine OCC were cultured in vitro in a medium with FSH/LH in the presence/absence of lapatinib.
MAIN OUTCOME MEASURE(S): Methods performed: real-time reverse transcriptase-polymerase chain reaction (PCR), immunofluorescence, RIA.
RESULT(S): In FSH/LH-stimulated and expanded cumulus oophorus extracellular matrix, HA was detected with biotinylated HA-binding proteins. However, weaker HA- and weaker cytoplasmic TNFAIP6 were detected were detected in lapatinib-pretreated OCC. The expression of the two cumulus expansion-associated gene transcripts was significantly decreased and synthesis of HA by cumulus cells was reduced. Lapatinib (10 μM) inhibited FSH/LH-induced oocyte meiotic maturation. Progesterone production increased after OCC stimulation with FSH/LH and was significantly decreased by lapatinib (10 μM).
CONCLUSION(S): Lapatinib inhibits oocyte maturation and reduces expression of cumulus expansion-associated transcripts, and synthesis of HA and P in OCC cultured in vitro in FSH/LH-supplemented medium.
研究表皮生长因子（EGF）受体酪氨酸激酶抑制剂拉帕替尼（lapatinib）对猪卵丘-卵母细胞复合体（OCC）中卵母细胞成熟、肿瘤坏死因子α诱导蛋白 6（TNFAIP6）和前列腺素内过氧化物合酶 2（PTGS2）等与卵丘扩展相关基因的表达以及透明质酸（HA）和孕酮（P）合成的影响。
本研究聚焦于拉帕替尼，这是一种口服活性的小分子，能选择性抑制 EGF 受体和人表皮生长因子受体 2 的酪氨酸激酶结构域及其下游信号转导。
生殖生物学实验室。
患者/人群：无。
在含有 FSH/LH 的培养基中，用或不用 lapatinib 对猪 OCC 进行体外培养。
实时逆转录-聚合酶链反应（PCR）、免疫荧光、放射免疫分析。
在 FSH/LH 刺激和扩展的卵丘-卵母细胞细胞外基质中，用生物素化的 HA 结合蛋白检测到 HA。然而，在 lapatinib 预处理的 OCC 中，HA 及细胞质中 TNFAIP6 表达减弱。两种与卵丘扩展相关的基因转录物的表达显著降低，且卵丘细胞合成的 HA 减少。拉帕替尼（10 μM）抑制 FSH/LH 诱导的卵母细胞减数分裂成熟。FSH/LH 刺激 OCC 后，P 生成增加，而 lapatinib（10 μM）显著降低了 P 的生成。
拉帕替尼抑制卵母细胞成熟，并降低了在 FSH/LH 补充培养基中体外培养的 OCC 中与卵丘扩展相关的基因转录物的表达，以及 HA 和 P 的合成。
