CORELATION BETWEEN HEPATIC SINUSOIDAL ENDOTHELIAL CELLS AND HUMAN COLORECTAL CARCINOMA CELLS IN LIVER METASTASIS

Liver是彩色肉瘤（CRC）的Metastasis的主要站点，并且可能有不同的机制来抑制CRC增长。我们已经使用了一种肿瘤细胞-金黄色内皮细胞（SEC）的Coculture系统，以评估SEC对细胞毒性较弱或较强的metastatic CRC细胞。SEC从正常瑞士老鼠的幸存者中分离出来，并被一种门户血管酶增流方法作为单体安装在96孔微滴定板上。Confluent SEC单体包含93%的内皮细胞（由低密度脂蛋白（LDL）受体抑制）和7%的Kupffer细胞。当CRC细胞预先标记为维生素颗粒的红胺-右旋糖胺和钙胺，然后用不饱和的SEC单体体进行鉴定的可行性，克隆细胞的%代谢活动，一个脆弱的Metastatic CRC，显著地低于CX-1细胞，一个高度Metastatic CRC，在培养4小时后（p<0.05）。Pretreatment of SEC gadolinium chloride （GaCl-D23-D2）， an inhibitor of KupOracle cell function， did not block the effect of SEC on Clone A cell。在24小时后，Coculture with Clone A， SEC生产了114±5.0 nM nitrite vs 123±5.0和8±2.0的SEC和Clone A细胞单独培养，因此，相应地。Furthermore，当10岁D1-6岁D1至1 mM N-D1 G-D1-甲基-L-精氨酸（NMMA）添加到SEC和克隆的混合物中，克隆一个细胞的毒性被阻断为硝酸生产被抑制超过10岁D1-2岁D1 mM。未刺激的骨髓SEC对较弱的金属克隆一个细胞具有毒性。因此， Hepatic SEC可能是一个主要宿主效应器细胞的种群，即通过较弱的Metastatic CRC，通过硝酸氧化物的生产。

The liver is the major site for metastasis by colorectal carcinoma (CRC) and may have different mechanisms that inhibit CRC growth. We have used a tumor cell-sinusoidal endothelial cell (SEC) coculture system to evaluate whether SEC are cytotoxic to weakly or highly metastatic CRC cells. SEC were isolated from the livers of normal Swiss mice by a portal vein enzymatic infusion method and established as monolayers in 96 well microtiter plates. Confluent SEC monolayers contained 93% endothelial cells (by low density lipoprotein (LDL) receptor staining) and 7% Kupffer cells. When CRC cells were prelabeled with the vital fluorescent dyes rhodamine-dextran and calcein AM and then cocultured with confluent SEC monolayers to assess viability, the % metabolic activity of Clone A cells, a weakly metastatic CRC, was significantly lower than that of CX-1 cells, a highly metastatic CRC, after 4 hrs of coculture (p<0.05). Pretreatment of SEC gadolinium chloride (GaClィイD23ィエD2), an inhibitor of Kupffer cell function, did not block the effect of SEC on Clone A cell. After 24 hrs of coculture with Clone A, SEC produced 114±5.0 nM nitrite vs 123±5.0 and 8±2.0 for SEC and Clone A cells cultured alone, respectively. Furthermore, when 10ィイD1-6ィエD1 to 1 mM NィイD1GィエD1-methyl-L-arginine (NMMA) was added to SEC and Clone A cocultures, toxicity to Clone A cells was blocked as nitrite production was inhibited by dose greater than 10ィイD1-2ィエD1 mM. Unstimulated murine SEC are more toxic to the weakly metastatic Clone A cells than highly metastatic CX-1 cells. Thus, hepatic SEC may be a major host effector cell population that eliminates weakly metastatic CRC through the production of nitric oxide.