DEVELPOMEN TO NEW TREATMENT AND DETAILS OF THE GENETIC CHANGES UNDERLYING THE NEOPLASTIC TRANSFORMATION OF UTERINE SMOOTH MUSCLE

平滑肌肉瘤仅占子宫恶性肿瘤的1.3%和子宫肉瘤的四分之一，但它是重要的，因为它是一种非常恶性的肿瘤，具有高的局部复发和远处转移率。由于放疗和化疗均无效，因此迫切需要一种新的治疗子宫平滑肌肉瘤的方法。然而，子宫平滑肌细胞肿瘤性转化的遗传变化的细节尚未得到充分的表征。Calponin hl是一种肌动蛋白结合蛋白，能够抑制平滑肌收缩，是平滑肌细胞（SMC）的组成成分。在子宫平滑肌肿瘤中，平滑肌肉瘤表现出钙调蛋白h1表达减少，正如我们以前报道的。因此，通过将其基因转染平滑肌肉瘤细胞来评估钙调蛋白hl的作用。将人钙调蛋白h1 cDNA构建体转染人平滑肌肉瘤细胞， ...更多信息 SKN和SK-LMS-1细胞系。为了证实新霉素抗性克隆中的基因转移和calponin h1蛋白的表达，通过Southern blotting、RT-PCR、Western blotting和免疫组织化学检测转染的细胞。通过MTT法、非贴壁生长实验和裸鼠成瘤实验，观察转染细胞的形态学变化，评价转染细胞的增殖活性和成瘤性。钙调蛋白hl转染细胞的细胞排列与培养的正常子宫肌层平滑肌细胞相似。转染人calponin h1基因的平滑肌肉瘤细胞在SK-LMS-1中的增殖抑制率为对照组的69%，在SKN中为70%。在锚定非依赖性生长和裸鼠致瘤性试验中，转钙调蛋白h1基因的平滑肌肉瘤细胞的生长和致瘤性均显著降低。钙调蛋白h1可能在平滑肌肉瘤中起抑癌作用。在临床上，通过将calponin h1基因导入低分化平滑肌肉瘤细胞中诱导正常分化表型可能具有潜在的治疗价值。少

Leiomyosarcoma constitutes only about 1.3% of uterine malignancies and one-quarter of uterine sarcomas, but it is important because it is an extremely malignant neoplasm with high rates of local recurrence and distant metastasis. The introduction of a new therapy for leiomyosarcoma of the uterus is urgently needed since neither radiotherapy nor chemotherapy is effective. However, the details of the genetic changes underlying the neoplastic transformation of uterine smooth muscle cells have not been fully characterized. Calponin hl, an actin-binding protein capable of inhibiting smooth muscle contraction, is a constitutive element of smooth muscle cells (SMCs). Among smooth muscle neoplasms of the uterus, however, leiomyosarcomas exhibit a reduced expression of calponin h1, as we reported previously. Therefore, the effect of calponin hl was assessed by transfecting its gene into leiomyosarcoma cells. A human calponin h1 cDNA construct was transfected into the human leiomyosarcoma cell l … More ines SKN and SK-LMS-lby electroporation. To confirm the gene transfer and expression of calponin h1 protein in the neomycin-resistant colonies, transfected cells were investigated by Southern blotting, RT-PCR, Western blotting and immunohistochemistry. We observed the morphology of calponin hl-transfected cells and evaluated their proliferative activity and the tumorigenicity by 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyl-2H-tetrazolium bromide (MTT) assay, anchorage-independent growth assay, and nude mice tumorigenicity assay. The cellular arrangement of calponin hl-transfected cells came to resemble that seen in cultured normal myometrial smooth muscle cells. The proliferation of human calponin h1-transfected leiomyosarcoma cells was inhibited to 69% of control in SK-LMS-1, and to 70% in SKN. In assays of anchorage-independent growth and nude mice tumorigenicity, growth and tumorigenicity were both significantly reduced in calponin h1-transfected leiomyosarcoma cells. Calponin h1 may function as a tumor suppressor in leiomyosarcoma. Clinically, induction of the normal differentiated phenotype by transferring the calponin h1 gene into poorly differentiated leiomyosarcoma cells may be of potential therapeutic value. Less