Yokomuro S, Tsuji H, Lunz J G, Sakamoto T, Ezure T, Murase N, Demetris A J
Thomas E Starzl Transplantation Institute, Division of Transplantation, University of Pittsburgh Medical Center, Pittsburgh, PA 15261, USA.
Hepatology. 2000 Jul;32(1):26-35. doi: 10.1053/jhep.2000.8535.
A well characterized human cholangiocarcinoma (CC) cell line, SG231, was compared with primary cultures of normal human biliary epithelial cells (BECs) for alterations in interleukin 6 (IL-6) and hepatocyte growth factor (HGF)-mediated stimulation and transforming growth factor beta1 (TGF-beta1) and activin A-mediated inhibition of growth. Results were compared with immunolabeling of the original tumor and after injection of SG231 into the liver of BALB/cByJ-scid mice. In vitro, both BECs and CCs expressed met, gp80, and gp130 messenger RNA (mRNA) and protein, but the levels of expression were higher in the CCs than in the BECs. In both the CCs and BECs, exogenous HGF or IL-6 induced phosphorylation of met or gp130, respectively, and a concentration-dependent increase in DNA synthesis. However, the CCs but not BECs, continued to grow in basal serum-free medium (SFM) and spontaneously produced both IL-6 and HGF under these conditions, which resulted in auto-phosphorylation of gp130 and met, respectively; and neutralizing anti-HGF or anti-IL-6 alone inhibited CC growth, indicative of autocrine growth control circuits. Conversely, activin A inhibits the growth of both BECs and CCs, but does not significantly increase apoptosis. Activin-A-induced growth inhibition of both CCs and BECs can be reversed by 100 ng/mL exogenous IL-6, but not by 10 to 100 ng/mL HGF. TGF-beta1 inhibited the growth of BECs but had no mitoinhibitory or proapoptotic effects on CCs. Immunolabeling of the original tumor and after inoculation into scid mice showed positive staining for met, gp130, gp80, and IL-6. This study contributes to a further understanding of BEC growth control and derangements that can occur during cholangiocarcinogenesis.
将一种特征明确的人胆管癌细胞系SG231与正常人胆管上皮细胞（BEC）的原代培养物进行比较，观察白细胞介素6（IL-6）和肝细胞生长因子（HGF）介导的刺激以及转化生长因子β1（TGF-β1）和激活素A介导的生长抑制方面的变化。将结果与原发肿瘤以及将SG231注射到BALB/cByJ-scid小鼠肝脏后的免疫标记结果进行比较。在体外，BEC和胆管癌细胞（CC）均表达met、gp80和gp130信使核糖核酸（mRNA）及蛋白质，但CC中的表达水平高于BEC。在CC和BEC中，外源性HGF或IL-6分别诱导met或gp130磷酸化，并导致DNA合成呈浓度依赖性增加。然而，CC在基础无血清培养基（SFM）中持续生长，且在这些条件下自发产生IL-6和HGF，分别导致gp130和met的自磷酸化；单独使用中和性抗HGF或抗IL-6可抑制CC生长，表明存在自分泌生长控制回路。相反，激活素A抑制BEC和CC的生长，但不会显著增加细胞凋亡。激活素A诱导的CC和BEC生长抑制可被100 ng/mL外源性IL--6逆转，但不能被10至100 ng/mL HGF逆转。TGF-β1抑制BEC生长，但对CC无促有丝分裂抑制或促凋亡作用。原发肿瘤以及接种到scid小鼠后的免疫标记显示met、gp130、gp80和IL-6呈阳性染色。本研究有助于进一步了解BEC生长控制以及胆管癌发生过程中可能出现的紊乱情况。
