肿瘤细胞治疗研究
英文原题：Human urine stem cells protect against cyclophosphamide-induced premature ovarian failure by inhibiting SLC1A4-mediated outflux of intracellular serine in ovarian granulosa cells.
我们的研究表明，基于 hUSC 的细胞疗法或简单补充丝氨酸可能为临床上预防和治疗 CTX 诱导的 POF 提供一种高效的治疗途径。
环磷酰胺（CTX）是治疗乳腺癌的一线药物，但它可能导致严重的卵巢功能障碍，甚至卵巢早衰（POF）。然而，CTX 诱导 POF 的机制仍不清楚。基于间充质干细胞的疗法已被广泛用于治疗多种疾病。因此，我们的研究旨在阐明 CTX 诱导 POF 的潜在机制，并探讨人尿源干细胞（hUSCs）在 POF 中的治疗作用。
本研究通过体外和体内实验，采用hUSCs及其外泌体处理CTX诱导的POF或卵巢颗粒细胞（GCs）凋亡。使用多种方法检测形态学、组织学和功能改变。通过卵巢差异表达分析确定hUSC来源外泌体（hUSC-Exo）的效应分子。通过GCs转录组测序获取miRNA的靶基因，并进一步阐明其潜在机制。
hUSCs 分别显著抑制了 CTX 诱导的凋亡并促进了 GCs 的增殖。此外，我们观察到 miR-27b-3p 在 hUSC-Exo 中高表达，并通过特异性抑制卵巢 GCs 中丝氨酸转运体 SLC1A4 的表达，显著抑制 CTX 诱导的 GC 凋亡，进而通过抑制细胞丝氨酸外流提高细胞内丝氨酸浓度。更重要的是，敲低 SLC1A4 或单纯补充丝氨酸均可抑制 CTX 诱导的 GCs 凋亡。最后，我们证明，CTX 通过上调 SLC1A4 的表达降低细胞内丝氨酸浓度，从而诱导卵巢 GCs 凋亡，而这对 POF 至关重要；而 hUSCs 则通过 miR-27b-3p/SLC1A4/丝氨酸轴介导的 PI3K/AKT/mTOR 信号通路激活，对 CTX 诱导的 POF 发挥保护作用。
BACKGROUND: Cyclophosphamide (CTX) is the first-line medication for the treatment of breast cancer, although it potentially leads to severe ovarian dysfunction and even premature ovarian failure (POF). However, the mechanism of CTX-induced POF remains unclear. Mesenchymal stem cell-based therapy has been wildly used for treating numerous diseases. Therefore, our study aims to elucidate the underlying mechanism of CTX-induced POF and to explore the therapeutic effect of human urine stem cells (hUSCs) in POF. METHODS: CTX-induced POF or ovarian granulosa cell (GCs) apoptosis were treated with hUSCs and their exosomes in vitro and in vivo. Morphological, histological, and functional alternations were examined using multiple approaches. The effector molecules of hUSC-derived exosomes (hUSC-Exo) were determined by differential expression analysis in the ovaries. The target genes of miRNA were accessed by transcriptome sequencing in GCs, and the underlying mechanisms were further elucidated. RESULTS: hUSCs remarkably inhibited CTX-induced apoptosis and promoted the proliferation of GCs, respectively. In addition, we observed that miR-27b-3p was highly expressed in hUSC-Exo and markedly suppressed CTX-induced GC apoptosis by specifically inhibiting the expression of SLC1A4, a serine transporter, in ovarian GCs, which, in turn, elevated the concentration of the intracellular serine by inhibiting the outflux of cellular serine. More importantly, the knockdown of SLC1A4 or simple supplementation of serine suppressed CTX-induced apoptosis of GCs. Finally, we demonstrated that CTX-induced apoptosis of ovarian GCs was essential for POF by reducing the intracellular serine concentration via elevating the expression of SLC1A4, whereas hUSCs protected against CTX-induced POF via miR-27b-3p/SLC1A4/serine axis-mediated activation of the PI3K/AKT/mTOR signaling pathway. CONCLUSIONS: Our study suggests that hUSC-based cell therapy or simple supplementation of serine may provide an efficient therapeutic approach for the prevention and treatment of CTX-induced POF clinically.
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