Possible role of calponin h1 as a tumor suppressor in human uterine leiomyosarcoma
Possible role of calponin h1 as a tumor suppressor in human uterine leiomyosarcoma
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DOI：
10.1093/jnci/91.9.790
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发表时间：
1999-05-05
期刊：
影响因子：
--
通讯作者：
Fujii, S
中科院分区：
文献类型：
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作者：
Horiuchi, A;Nikaido, T;Fujii, S
背景：钙调蛋白h1是一种能够抑制平滑肌收缩的碱性肌动蛋白结合蛋白，是平滑肌细胞的组成元件。然而，正如我们之前报道的，在平滑肌肉瘤（子宫平滑肌肿瘤的一种）中，观察到钙调蛋白 h1 表达减少。在这项研究中，我们试图评估增加平滑肌肉瘤细胞中钙调蛋白 hi 表达的影响（体外和体内）。方法：将含有人钙调蛋白hi互补DNA和细菌新霉素抗性基因的质粒转染人平滑肌肉瘤细胞系SKN和SK-LMS-1，通过电穿孔、Southern印迹、逆转录聚合酶链反应分析、蛋白质印迹和免疫组化来证实钙调蛋白hi蛋白的DNA转移和表达。 新霉素抗性克隆。我们表征了钙调蛋白hl转染细胞的形态，并通过使用3-(4,5-二甲基噻唑-2-基)-2,5-二苯基-2H-四唑溴化物测定、不依赖贴壁的生长测定和裸鼠致瘤性测定评估了它们的增殖活性和致瘤性。结果：培养中转染钙调蛋白h1的细胞的形态与培养的正常子宫肌层平滑肌细胞的形态相似。对于SK-LMS-1细胞，钙调蛋白hl转染细胞的增殖减少至对照的69%；对于 SKN 细胞，calponin hi 转染将增殖降低至对照的 70%。在贴壁依赖性生长和体内致瘤性测定中，钙调蛋白 h1 转染的平滑肌肉瘤细胞的生长和致瘤性均统计显着降低。结论：钙调蛋白 hi 可能在平滑肌肉瘤中发挥肿瘤抑制作用。临床上，将钙调蛋白 h1 互补 DNA 转移到低分化的平滑肌肉瘤细胞中，通过诱导正常的分化细胞表型，可能具有潜在的治疗价值。
Background: Calponin h1, a basic actin-binding protein capable of inhibiting smooth muscle contraction, is a constitutive element of smooth muscle cells. However, in leiomyosarcoma (a type of smooth muscle neoplasm of the uterus), reduced expression of calponin h1 is observed, as we have reported previously. In this study, we sought to assess the effects (in vitro and in vivo) of increasing calponin hi expression in leiomyosarcoma cells. Methods: A plasmid containing a human calponin hi complementary DNA and a bacterial neomycin-resistance gene was transfected into the human leiomyosarcoma cell lines SKN and SK-LMS-1 by electroporation, Southern blotting, reverse transcription-polymerase chain reaction analysis, western blotting, and immunohistochemistry were used to confirm DNA transfer and expression of the calponin hi protein in neomycin-resistant clones. We characterized the morphology of calponin hl-transfected cells, and we evaluated their proliferative activity and tumorigenicity by use of a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide assay, an anchorage-independent growth assay, and a nude mouse tumorigenicity assay. Results: The morphology of calponin h1-transfected cells in culture resembled that of cultured normal myometrial smooth muscle cells. With SK-LMS-1 cells, proliferation of calponin hl-transfection cells was reduced to 69% of control; with SKN cells, calponin hi transfection reduced proliferation to 70% of control. In assays of anchorage-independent growth and in vivo tumorigenicity, both growth and tumorigenicity were statistically significantly reduced in calponin h1-transfected leiomyosarcoma cells. Conclusions: Calponin hi may function as a tumor suppressor in leiomyosarcoma, Clinically, transfer of a calponin h1 complementary DNA into poorly differentiated leiomyosarcoma cells may be of potential therapeutic value through induction of a normal, differentiated cellular phenotype.