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目的 :本研究旨在评价反义寡聚脱氧核糖核苷酸 (ODN)对促卵泡刺激素受体 (FSHR)mRNA表达的调节作用。 方法 :转染FSHR的CHO细胞系的mRNA水平用反转录 竞争性PCR方法进行定量测定。 结果 :结果显示用 10 μmol/L反义ODN可导致mRNA表达显著上升 ,2 4h后mRNA水平从 0 89± 0 0 6pg/ μg总RNA升至 1 6 4±0 0 8pg/ μg总RNA(P <0 0 5 )。用 0 33μmol/L反义ODN进行脂转染也得到相同结果 ,mRNA水平从 0 95± 0 0 8pg/ μg总RNA升高至 1 5 3± 0 0 7pg/ μg总RNA。这可能是由于FSHR蛋白翻译被抑制后上行调节mRNA合成所致 ,无义ODN对FSHRmRNA的表达没有任何作用。 结论 :本文结果提示针对猪FSHR翻译起始位点的反义ODN能上行调节猪FSHRmRNA的表达水平。
Aims: This study aimed to evaluate the regulatory effect of antisense oligodeoxyribonucleotides (ODNs) on follicle stimulating hormone receptor (FSHR) mRNA expression. Methods: The mRNA levels of CHO cell lines transfected with FSHR were quantitatively determined using a reverse transcription competitive PCR method. Results: The results showed that mRNA expression increased significantly with 10 μmol / L antisense ODN. After 24 h, the mRNA level increased from 0 89 ± 0 6 pg / μg total RNA to 1 64 ± 0 0 8 pg / μg total RNA (P <0 0 5). The same result was obtained by lipofection with 0 33 μmol / L antisense ODN, with mRNA levels increasing from 0 95 ± 0 0 pg / μg total RNA to 153 ± 0 0 7 pg / μg total RNA. This may be due to up-regulated mRNA synthesis after FSHR protein translation is inhibited, and nonsense ODN has no effect on FSHR mRNA expression. CONCLUSIONS: Our results suggest that antisense ODN directed against the translation initiation site of porcine FSHR can upregulate the expression of porcine FSHR mRNA.