子宮肌瘤是好發於生育年齡婦女的良性腫瘤，其為造成婦女接受切除子宮手術之主要病因之一。肌瘤表現大量的細胞外基質及轉化生長因子，轉化生長因子能促進子宮肌瘤細胞的細胞外基質表現以及細胞增生。前人研究發現子宮肌瘤細胞經長時間培養後可形成三維立體的球狀聚集。本研究藉由長時間細胞培養方式來探討乙型轉化生長因子對子宮肌瘤細胞形成球狀聚集之影響。子宮肌瘤細胞取自於進行子宮全切除術病患之肌瘤組織。肌瘤細胞經培養七天或十四天會形成不同程度之細胞聚集。子宮肌瘤細胞聚集程度可分為Type-A、Type-B和Type-C。Type-A為低密度而交錯之多層細胞聚集，Type-B為高密度多層細胞聚集，Type-C為高密度球狀細胞聚集。子宮肌瘤細胞比子宮平滑肌細胞較易形成細胞聚集。我們的實驗結果顯示子宮肌瘤細胞於0.5 %和2 % FBS-DMEM培養下，轉化生長因子皆可以促進細胞聚集。過去文獻指出子宮肌瘤細胞會分泌轉化生長因子，我們因此利用第一型轉化生長因子受體抑制劑(SB431542)來抑制轉化生長因子接受器的磷酸化，進而抑制轉化生長因子所引發之Smad-3活化，SB431542可抑制子宮肌瘤細胞聚集的過程。U0126是一種抑制ERK 磷酸酶的抑制劑，它抑制肌瘤細胞ERK的磷酸化，並且會抑制子宮肌瘤細胞形成聚集的過程。總結，在細胞聚集的過程中，轉化生長因子可以誘導Smad-3的磷酸化進而促進子宮肌瘤細胞的聚集，此外ERK所傳遞的訊息也參與在細胞聚集的過程。

Leiomyoma, a benign tumor derived from myometrium, is the most common gynecological problem to cause hysterectomy. Previous studies have shown that leiomyoma expresses higher levels of TGF-beta and extracellular matrix compared to myometrium. In addition, leiomyomal cells form three dimensional aggregates after long-term culture. This study aimed to investigate the role of TGF-βin leiomyomal cell aggregation in long-term culture. Leiomyomal cells from patients underwent hysterectomy were cultured for 7 or 14 days. Multilayered cell aggregates were classified according to the cell density detected under phase-contrast optics: type A, low-density aggregates; type B, high-density aggregates; type C, high-density spheroids. TGF-β enhanced the aggregation of leiomyomal cells cultured in DMEM containing 0.5% or 2% fetal bovine serum. SB431542, an inhibitor for TGF-βreceptor kinase, inhibited TGF-β-enhanced aggregation in leiomyomal cells. The phosphorylation of smad-3 was stimulated by TGF-βin a time-dependent manner, with peak activation at 1 hr. SB431542 eliminated the TGF-β-induced smad-3 phosphorylation. U0126, an inhibitor for ERK kinase, inhibited ERK phosphorylation and aggregate formation in leiomyomal cells. These results suggested that TGF-β/smad 3and ERK signaling pathways are involved in leiomyomal cell aggregation.

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