子宮肌瘤在生育年齡的婦女中是最常見的良性腫瘤，相關的症狀包括子宮大量不正常的出血，不孕，以及反覆性的流產等等。先前的研究指出相較於正常的子宮肌膜層，子宮肌瘤表現出較高的 TGF-β3。 TGF-β 可參與調控細胞外基質的產生以及細胞增生。先前的研究者觀察到從子宮肌瘤組織中所得到的子宮肌瘤細胞，在經過三個星期的培養後，會形成一個圓球狀的結構 (ball-like structure)。這些圓球狀的結構與人體內的子宮肌瘤十分相像，並且正常的子宮肌膜層無法形成這樣的結構。我們利用這樣的結構當作子宮肌瘤的體外研究模型，來探討TGF-β 在子宮肌瘤形成過程中所扮演的角色。子宮肌瘤組織是從在成大醫院進行子宮全切術的病患身上取得，而後子宮肌瘤細胞會自子宮肌瘤組織中取得，並培養於含有2% 胎牛血清的培養液。經過約四星期的培養後，細胞會形成不同類型的圓球狀結構，我們將其稱為spheroid。利用phase-contrast顯微鏡技術以及Hoechst staining免疫螢光染色，我們根據spheroid的外型和細胞聚集的程度將spheroid區分為1-3三種類型。結果顯示 TGF-β3會促進後期 (type 3) spheroid的形成。SB431542是一種TGF-β訊息傳遞的阻斷者，它則會抑制type 1-3 spheroid的形成。另外，我們更進一步檢測細胞的排列是否會影響到spheroid的形成。藉由肌動蛋白的免疫螢光染色，我們發現 TGF-β3 會使細胞的排列更加不規則，相對的， SB431542則會使細胞以相互平行的形式排列。由以上我們可得知TGF-β 對於spheroid的形成扮演重要的角色，但對於它如何影響細胞的排列，進而影響spheroid的生成則需要更進一步的探討。

Uterine leiomyomas are the most common benign tumors in women of reproductive age. The symptoms associated with leiomyomas include excessive uterine bleeding, infertility and repetitive pregnancy loss. Previous studies have shown that uterine leiomyomas express higher level of TGF-β3 and extracellular matrix compared with normal myometrium. TGF-βs are shown to participate in extracellular matrix production and cell proliferation. Previous investigators have observed that leiomyoma cells from tissue explants formed ball-like aggregates after 3-week culture. These ball-like aggregates resembled the nodules of leiomyoma in vivo. Myometrial cells did not form such structures. Using this model, we have investigated the roles of TGF-β in leiomyoma development. Leiomyomal tissues were obtained from the patients who underwent hysterectomy at the NCKU hospital. Cells were isolated from tissue explants and cultured in 2% FCS. After 4 weeks, different types of ball-like aggregates formed. We called them spheroids. The spheroids were classified as type 1 to type 3 according to the morphology of spheroids and the extent of cell aggregation by phase-contrast microscopy and Hoechst staining. The quantitative results showed that growth rate of spheroids varied in leiomyomal cells from different tumors. Our results showed that TGF-β3 treatment promoted the formation of advanced types of spheroids. However, SB431542, a TGF-β signalling blocker reduced the formation of type 1-3 spheroids. We further investigated whether cellular arrangement was involved in spheroid development. The immunostaining of F-actin revealed that TGF-β3 promoted disorganized arrangement of leiomyomal cells. In contrast, cells grew in parallel under SB431542 treatment. All together, our results suggest that TGF-β may play a role in spheroid development. The mechanism by which SB431452 reduces spheroid formation remains further investigation.

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