Tamai T, Patsch W, Schonfeld G
Department of Medicine, Washington University School of Medicine, St. Louis, MO 63110.
Atherosclerosis. 1988 Jan;69(1):29-37. doi: 10.1016/0021-9150(88)90286-9.
The rat H-35 cultured hepatoma cell line expresses receptors for homologous lipoproteins. In previously reported experiments distinct receptors were identified for chylomicron remnants, HDL and LDL, by direct binding studies that yielded distinctive binding constants, cross competition assays, and by differential inhibitory effects of EDTA and suramin. In the present experiments, the regulation of expression of these receptors was assessed by growing cells either in the presence or absence of lipoproteins in the media and by growing cells to different densities (50-800 micrograms cell protein/dish). LDL binding to cells was increased by lipoprotein deprivation at all cell densities. LDL binding was inversely related to cell density when cells were grown in lipoprotein deficient serum (LPDS) but cell density did not affect LDL binding by cells grown in newborn calf serum (NBCS). By contrast HDL binding was not appreciably different whether cells were grown in NBCS or in LPDS. However, HDL binding was inversely related to cell density by cells grown either in LPDS or in NBCS. Binding of chylomicron remnants was increased by growth in LPDS at all densities, but altering growth density in either culture medium had little effect on the cellular binding of chylomicron remnants. The distinctive effects of these experimental perturbations on the binding of the 3 lipoprotein classes tend to confirm the presence of 3 separate receptor activities. The experiments also demonstrate that the responses at least of some of the receptors of the hepatoma cells in culture resemble those of hepatocytes in vivo and in vitro.(ABSTRACT TRUNCATED AT 250 WORDS)
大鼠H-35培养肝癌细胞系表达同源脂蛋白受体。在先前报道的实验中,通过直接结合研究确定了乳糜微粒残粒、高密度脂蛋白(HDL)和低密度脂蛋白(LDL)的不同受体,该研究得出了独特的结合常数、交叉竞争试验,以及乙二胺四乙酸(EDTA)和苏拉明的差异抑制作用。在本实验中,通过在培养基中存在或不存在脂蛋白的情况下培养细胞,以及将细胞培养至不同密度(50 - 800微克细胞蛋白/培养皿)来评估这些受体表达的调节。在所有细胞密度下,脂蛋白剥夺均增加LDL与细胞的结合。当细胞在脂蛋白缺乏血清(LPDS)中生长时,LDL结合与细胞密度呈负相关,但细胞密度不影响在新生小牛血清(NBCS)中生长的细胞的LDL结合。相比之下,无论细胞在NBCS还是LPDS中生长,HDL结合均无明显差异。然而,在LPDS或NBCS中生长的细胞,HDL结合均与细胞密度呈负相关。在所有密度下,在LPDS中生长可增加乳糜微粒残粒的结合,但在任何一种培养基中改变生长密度对乳糜微粒残粒的细胞结合影响不大。这些实验干扰对这三类脂蛋白结合的独特作用倾向于证实存在三种独立的受体活性。实验还表明,培养的肝癌细胞中至少某些受体的反应类似于体内和体外肝细胞的反应。(摘要截短于250字)