Development of screening system for searching cellular differentiation-inducing agents by utlizing human salivary cancer cells

1.从5-氮杂胞苷处理的人唾液癌细胞系HSG中分离到具有丰富的神经丝束和内质网以及微管蛋白α和β链的神经元样细胞（HSG- azall）。发现HSG-AZAll细胞来自神经突，对二丁基环amp反应表达突触素和神经元特异性烯醇化酶，这表明HSG-AZAll细胞可能有助于寻找通过cAMP信号转导起作用的细胞分化诱导因子。2.我们发现，用5-氟脱氧尿苷单磷酸处理HSG细胞可分化为肌上皮细胞并诱导凋亡。3.我们发现，在3,4-二氢-6-[3,4-二甲氧基苯甲酰)-1-哌嗪基]-2 (1H) -喹诺酮（vesnarinone）存在的情况下，用5-氮杂胞苷处理HSG- aza3细胞，培养出具有腺泡细胞表型的HSG- aza3细胞，可分化为软骨细胞。此外，已有研究发现22-oxa-1 α， 25-二羟基维生素D_3处理HSG-AZA3细胞可诱导分化为成骨细胞。4.我们发现，用依托泊苷（DNA拓扑异构酶II的抑制剂）处理HSG细胞可诱导平滑肌细胞表型的细胞，如α -平滑肌肌动蛋白和肌丝的表达，以及诱导凋亡。5.来源于人口腔小唾液腺的腺样鳞状癌形成细胞（TYS）经5-氟尿嘧啶或vesnarinone处理后，发现其诱导细胞凋亡并分化为腺泡细胞。6.22-oxa-1 α, 25 (OH) _2D_3处理裸鼠异种移植的HSG-AZA3细胞，肿瘤生长明显受到抑制，可见骨形成和骨重塑。7.当用vesnarinone处理TYS裸鼠肿瘤时，在处理的肿瘤中检测到肿瘤生长明显抑制，并向角化细胞和腺泡细胞分化。少

1.Neuron-like cells (HSG-AZAll) with ample neurofilament bundles and endoplasmic reticulum as well as alpha-and beta-chain of tubulin were isolated from human salivary cancer cell line HSG treated with 5-azactidine. This HSG-AZAll cells were found to from neurites and express synaptophysin and neuron-specific enolase in response to dibutyryl cyclic AMP.This indicated that HSG-AZAll cells might be useful for searching cellular differentiation-inducing agents which work via cAMP signal transduction. 2.We have found that the treatment of HSG cells with 5-fluorodeoxyuridine monophosphate results in the differentiation into myoepithelial cells and induciton of apoptosis. 3.We have found that the cultivation of HSG-AZA3 cells with an acinar cell phenotype, which were induced by treatment of HSG cells with 5-azacytidine, in the presence of 3,4-dihydro-6-[3,4-dimethoxybenzoyl)-1-piperazinyl]-2 (1H) -quinolnone (vesnarinone) results in the differentiation into chondrocytes.In addition, it has b … More een found that the treatment of HSG-AZA3 cells with 22-oxa-1alpha, 25-dihydroxyvitamin D_3 causes the differentiation into osteoblasts. 4.We have found that the treatment of HSG cells with etoposide, an inhibitor of DNA topoisomerase II,results in the induction of cells with a smooth muscle cell phenotype, such as expression of alpha-smooth muscle actin and myofilaments, as well as in the induction of apoptosis. 5.When adenoid squamous carcinoma-forming cells (TYS) , derived from human minor salivary gland present in the oral cavity, was treated with 5-fluorouracil or vesnarinone, it has been found that the induction of apoptosis and differentiation into acinar cells occurs in the treated cells. 6.The treatment with 22-oxa-1alpha, 25 (OH) _2D_3 of HSG-AZA3 cells grown as xenografts in nude mice resulted in the marked growth inhibition of tumors, in which bone formation and bone remodeling were detected. 7.When TYS nude mouse tumors were treated with vesnarinone, significant suppression of tumor growth as well as differentiation into both keratinocytes and acinar cells were detected in the treated tumors. Less