趋化因子MDC/CCL22-CCR4反应轴在小鼠胃癌腹膜乳斑转移中的作用

目的 探讨趋化因子MDC/CCL22-CCR4反应轴在小鼠胃癌腹膜乳斑转移中的作用.方法 用RT-PCR和Western blot方法检测615小鼠前胃癌细胞(MFC)CCR4的mRNA和蛋白质的表达;在615实验小鼠腹腔内注射0.2 ml Dil荧光颗粒标记的MFC悬液(含癌细胞1×104个)制备腹膜乳斑转移模型,观察MFC转移过程及部位,取大网膜分别做CCL22和CCR4的免疫组化染色,在扫描电镜下观察乳斑区结构.实验动物被分为MFC组和生理盐水组,在腹腔注入MFC后第6、8、10天用ELISA方法分别检测MFC组和生理盐水对照组腹水中的CCL22浓度. 结果 MFC初期转移在大网膜乳斑区,在乳斑区可检测到CCR4和CCL22表达.在扫描电镜下可见乳斑区主要由巨噬细胞、淋巴细胞和不连续的内皮细胞构成.CCL22在盐水对照组平均值是43 pg/ml,MFC组平均值是364 pg/ml.结论 MDC/CCL22-CCR4反应轴在小鼠胃癌大网膜乳斑转移中可能发挥重要作用.

Role of MDC/CCL22-CCR4 axis in mouse milky spots with peritoneal carcinomatosis of gastric cancer

Objective To study role of MDC/CCI22-CCR4 axis in mouse milky spots with peritoneal carcinomatosis of gastric cancer. Methods We examined the expression of CCR4 in 615 Mouse gastric cancer cell (MFC) lines by RT-PCR and Western-blot; Peritoneal metastasis model on the 615mouse was established by intraperitoneal injection of 0.2 ml MFC cells(1×104 cells). Dil fluorescence was used to observe the transfer process and section of MFC. Immunohistochemistry was conducted to detect the expression of CCR4 and CCL22 in omental milky spot; the structure of Milky spot was observed by scanning electron microscopy. Mice were randomly divided into 2 groups, namely, the saline control group (received saline) and MFC group. The concentration of CCL22 in ascitic fluid was measured in the 615 mice injected MFC after 6,8,10 days and in the saline group. Results MFC first metastasizes to the milky spot on the omentum, the expression of CCR4 and CCL22 were observered in the milky spot. The surface layer cells in milky spot consisted of discontinuous mesothelial cells and mainly macrophages and lymphocytes. The average value of CCL22 was 43 pg/ml and 364 pg/ml respectively in saline control group and MFC group.Conclusions MDC/CCL22-CCR4 axis plays an important role in the development of peritoneal carcinomatosis in mouse gastric cancer.