Human leiomyoma smooth muscle cells show increased expression of transforming growth factor-β3 (TGFβ3) and altered responses to the antiproliferative effects of TGFβ
Human leiomyoma smooth muscle cells show increased expression of transforming growth factor-β3 (TGFβ3) and altered responses to the antiproliferative effects of TGFβ
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DOI：
10.1210/jc.86.2.913
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发表时间：
2001-02-01
影响因子：
5.8
通讯作者：
Nowak, RA
中科院分区：
文献类型：
--
作者：
Lee, BS;Nowak, RA
转化生长因子- β (TGF β)是调节多种细胞生长和分化的多功能肽。本研究的目的是比较正常子宫肌层和良性子宫平滑肌瘤肿瘤中TGF β亚型的表达，并在体外研究TGF β对这些细胞增殖和胶原生成的影响。子宫内膜和平滑肌瘤组织来自择期子宫切除术患者。对组织进行核糖核酸（RNA）处理，并建立原代细胞培养物。Northern blot分析显示，TGF - beta1 mRNA在肌瘤和子宫内膜之间表达水平相似，而平滑肌瘤的TGF - beta3 mRNA表达水平比自体子宫内膜高5倍。免疫组化检测TGF β 3蛋白在平滑肌瘤组织中的表达明显高于相应的肌层。在体外培养的平滑肌瘤和子宫肌平滑肌细胞中，TGF β 1和TGF β 3水平随细胞密度的增加而升高。通过氚化胸腺嘧啶掺入法测定DNA合成变化，评估TGF β 1和TGF β 3对细胞增殖的影响。TGF β的剂量分别为0、0.1、1.0、10.0 ng/mL。三种剂量的TGF β 1和TGF β 3对肌层平滑肌细胞DNA合成的抑制作用均为31-54%。与TGF β 1-3免疫中和抗体同时治疗可逆转这种抑制作用。TGF beta1对平滑肌瘤平滑肌细胞无影响，而TGF beta3增加平滑肌瘤细胞的DNA合成。与免疫中和抗体联合治疗可防止这种增加。用TGF β免疫中和抗体治疗平滑肌瘤和子宫肌瘤细胞24小时，可使I型和III型胶原mRNA水平降低45-60%，提示内源性TGF β对胶原生成很重要。这些结果支持了TGF β系统的改变导致对TGF β抗增殖作用的敏感性丧失的假设，TGF β 3表达的增加可能有助于这些肿瘤的生长。
Transforming growth factor-betas (TGF betas) are multifunctional peptides that regulate growth and differentiation in a variety of cells. The goals of this study were to compare expression of the TGF beta isoforms in normal myometrium and benign leiomyoma tumors of the uterus and to examine the effects of TGF betas on cell proliferation and collagen production by these cells in vitro. Myometrium and leiomyoma tissues were obtained from patients undergoing elective hysterectomies. Tissues were processed for ribonucleic acid (RNA) and were also established as primary cell cultures. Northern blot analysis showed that the levels of TGF beta1 messenger RNAs (mRNAs) were similar between leiomyoma and myometrium, whereas leiomyoma showed 5-fold higher levels of expression of TGF beta3 mRNA than autologous myometrium. Expression of TGF beta3 protein detected by immunohistochemistry was much more intense in leiomyoma tissues than in corresponding myometrium. Levels of both TGF beta1 and TGF beta3 increased with increasing cell density for leiomyoma and myometrium smooth muscle cells cultured in vitro. Effects of TGF beta1 and TGF beta3 on cell proliferation were assessed by measuring changes in DNA synthesis with the tritiated thymidine incorporation assay. The doses of TGF betas tested were 0, 0.1, 1.0, and 10.0 ng/mL. All three doses of TGF beta1 and TGF beta3 inhibited DNA synthesis in myometrium smooth muscle cells by 31-54%. Concomitant treatment with an immunoneutralizing antibody to TGF beta1-3 reversed this inhibitory effect. In contrast, TGF beta1 had no effect on leiomyoma smooth muscle cells, whereas TGF beta3 increased DNA synthesis by leiomyoma cells. Combined treatment with the immunoneutralizing antibody prevented this increase. Treatment of leiomyoma and myometrial cells with the TGF beta immunoneutralizing antibody for 24 h caused a 45-60% reduction in collagen type I and type III mRNA levels, suggesting that endogenous TGF betas are important for collagen production. These results support the hypothesis that alterations in the TGF beta system produce loss of sensitivity to the antiproliferative effects of TGF beta, and increased expression of TGF beta3 may contribute to the growth of these tumors.