Hishinuma A, Yamanaka T, Kasai K, So S, Tseng C C, Bamba N, Ohtake H, Shimoda S
Department of Endocrinology, Dokkyo University School of Medicine, Tochigi, Japan.
Thyroid. 1995 Feb;5(1):41-6. doi: 10.1089/thy.1995.5.41.
To study the growth control of human thyroid cells in different stages of differentiation, we established two human thyroid cell lines of adenomatous goiter and papillary carcinoma. A 59-year-old female patient with adenomatous goiter was operated in September 1991, and a 27-year-old female patient with papillary carcinoma in May 1990. The thyroid cell lines were established by successive passage without cellular or genetic manipulations such as fusing other cell lines or oncogenic viral infection. These cell lines, human adenomatous goiter cells (hAG) and human papillary thyroid carcinoma cells (hPTC), exhibited a flattened polygonal shape and proliferated as a monolayer in cell culture. The doubling time of the hAG cells was 60 h in Ham's F12 medium supplemented with 10% fetal bovine serum, and that of the hPTC cells, 18 h in the same medium. Both cell lines expressed mRNA for TSH receptor and secreted cAMP into the medium during incubation with thyrotropin (TSH) at concentrations as low as 0.01 mU/mL. The effects of activators of protein kinase A (PKA), protein kinase C (PKC), tyrosine kinase (TK), and estradiol (E2) on proliferation of the hAG cells and the hPTC cells were assessed by measuring cellular DNA content in 24-well plates with diaminobenzoic acid. TSH stimulated proliferation of the hAG cells, but it inhibited proliferation of the hPTC cells. Since TSH activates two signaling pathways, the adenyl cyclase-PKA system and phospholipase C-PKC system, we tested effects of dibutylyl cAMP (dBC) and phorbol myristate 13-acetate (PMA), separately. dBC stimulated proliferation of the hAG cells, but it inhibited that of the hPTC cells.(ABSTRACT TRUNCATED AT 250 WORDS)
为研究人甲状腺细胞在不同分化阶段的生长调控,我们建立了两株来源于腺瘤性甲状腺肿和乳头状癌的人甲状腺细胞系。1991年9月,对一名59岁患腺瘤性甲状腺肿的女性患者进行了手术,1990年5月,对一名27岁患乳头状癌的女性患者进行了手术。通过连续传代建立甲状腺细胞系,未进行诸如与其他细胞系融合或致癌病毒感染等细胞或基因操作。这些细胞系,人腺瘤性甲状腺肿细胞(hAG)和人乳头状甲状腺癌细胞(hPTC),呈扁平多边形,在细胞培养中以单层形式增殖。在补充有10%胎牛血清的Ham's F12培养基中,hAG细胞的倍增时间为60小时,hPTC细胞在相同培养基中的倍增时间为18小时。两株细胞系在与低至0.01 mU/mL的促甲状腺激素(TSH)孵育期间均表达TSH受体的mRNA并向培养基中分泌cAMP。通过用二氨基苯甲酸在24孔板中测量细胞DNA含量,评估蛋白激酶A(PKA)、蛋白激酶C(PKC)、酪氨酸激酶(TK)和雌二醇(E2)的激活剂对hAG细胞和hPTC细胞增殖的影响。TSH刺激hAG细胞增殖,但抑制hPTC细胞增殖。由于TSH激活两条信号通路,即腺苷酸环化酶 - PKA系统和磷脂酶C - PKC系统',我们分别测试了二丁酰环磷腺苷(dBC)和佛波醇12 - 肉豆蔻酸酯13 - 乙酸酯(PMA)的作用。dBC刺激hAG细胞增殖,但抑制hPTC细胞增殖。(摘要截短于250字)