Characterization of bone-marrow derived fibroblast recruited into cancer induced stroma.

众所周知，癌症-基质相互作用在癌症进展过程中发挥重要作用。将人胰腺癌细胞系Capan-1皮下异种移植到SCID受体中，并在第14天和第28天分析基质形成。免疫组织化学和免疫荧光研究显示，BM来源的内皮细胞（X-gal/CD 31或H-2b/CD 31双阳性细胞）和肌成纤维细胞（X-gal/α-平滑肌肌动蛋白或H-2b/α-平滑肌肌动蛋白双阳性细胞）存在于癌巢内和癌巢周围。第14天，BM来源的内皮细胞（BMD-VE）和BM来源的肌成纤维细胞（BMD-MF）的频率分别为25.3±4.4%和12.7± 9.6%。在第28天，BM衍生的内皮细胞的频率为26.7± 9.7%，这与第14天的值相似。但28 d时骨髓源性肌成纤维细胞的频率（39.8±17.1%）显著高于14 d（P<0.05）。拓扑异构酶Ⅱ α阳性率为2.2±1.2%， ...更多信息 与H-2b阴性肌成纤维细胞的0.3±0.4%相比，BM来源的肌成纤维细胞具有显著的增殖活性（P<0.05）。我们的研究结果表明，BM来源的肌成纤维细胞成为肿瘤发展后期癌症诱导的基质细胞的主要组成部分。此外，为了评估癌细胞类型的差异对BMD-VE和BMD-MF募集的影响，将10种人癌细胞系植入移植有双突变小鼠（RAG-1-/- b-gal Tg或RAG-1-/- GFP Tg）骨髓的免疫缺陷小鼠的皮下组织中。测量BMD-VE（%BMD-VE）和BMD-MF（%BMD-MF）的募集频率以及肿瘤相关参数：肿瘤体积（TV）、微血管密度（MVD）和间质比例（%St）。分析了它们之间的相关性。虽然%BMD-VE和%BMD-MF因细胞系不同而不同（分别为0 ~ 21.6%，0 ~ 29.6%），但这两个参数均与%St显著相关（P<0.005）。此外，%BMD-VE与%BMD-MF也有显著相关性（P<0.005）。为了评估肿瘤生长部位对目标细胞募集的影响，将人胰腺癌细胞系Capan-1移植到5个不同部位：皮下组织、腹膜、肝、脾和肺。皮下和腹膜肿瘤诱导结缔组织增生性间质（%St分别为22.7%，19.5%），含有BMD-VE（%BMD-VE= 21.6%，16.5%）和BMD-MF（%BMD-MF分别为29.6%，24.5%），但在肝、脾和肺的肿瘤中观察到弱的间质诱导，而无BMD-VE或-MF的募集（%St分别为9.7%，9.1%，5.4%）。这些结果表明，招聘的BMD-VE和-MF是需要在癌症进展过程中的基质形成，和癌症微环境是重要的招聘BMD-VE和-MF。少

Cancer-stromal interaction is well known to play important roles during cancer progression. Human pancreatic cancer cell line Capan-1 was subcutaneously xenotransplanted into SCID recipients and stromal formation was analyzed on day 14 and on day 28. Immunohistochemical and immunofluorescence studies revealed that BM-derived endothelial cells (X-gal/CD31 or H-2b/CD31 double-positive cells) and myofibroblasts (X-gal/α-smooth muscle actin or H-2b/α-smooth muscle actin double-positive cells) were present within and around the cancer nests. On day 14,the frequencies of BM-derived endothelial cells (BMD-VE) and BM-derived myofibroblasts (BMD-MF) were 25.3±4.4% and 12.7±9.6%, respectively. On day 28,the frequency of BM-derived endothelial cells was 26.7±9.7%, which was similar to the value on day 14. However, the frequency of BM-derived myofibroblasts was significantly higher (39.8±17.1%) on day 28 than on day 14 (P<0.05). The Topoisomerase IIα-positive ratio was 2.2±1.2% for the H-2b-positi … More ve myofibroblasts, as opposed to only 0.3±0.4% for the H-2b-negative myofibroblasts, significant proliferative activity was observed in the BM-derived myofibroblasts (P<0.05). Our results indicate that BM-derived myofibroblasts become a major component of cancer-induced stromal cells in the later stage of tumor development. Furthermore, to assess the effect of the difference in cancer cell types on the recruitment of BMD-VE and BMD-MF,10 kinds of human cancer cell line were implanted into the subctaneous tissue of immunodeficient mice transplanted with bone marrow of double-mutant mice (RAG-1-/- b-gal Tg or RAG-1-/- GFP Tg). The recruitment frequency of BMD-VE (%BMD-VE) and BMD-MF (%BMD-MF), and tumor-associated parameters : tumor volume(TV), microvessel density(MVD), and stromal proportion (%St) were measured. The correlation among them was analyzed. Although %BMD-VE and %BMD-MF varied (from 0 to 21.6%,0 to 29.6%, respectively) depending on the cancer cell line, both parameters significantly correlated with %St (P<0.005). Furthermore %BMD-VE and %BMD-MF also significantly correlated (P<0.005). In order to assess the effect of tumor growth sites on the recruitment of the cells of interest, a human pancreatic cancer cell line, Capan-1,was transplanted into 5 different sites : subcutaneous tissue, peritoneum, liver, spleen, and lung. Tumors in the subcutaneous tissue and peritoneum induced desmoplastic stroma (%St=22.7%,19.5%, respectively) and contained BMD-VE (%BMD-VE=21.6%,16.5% respectively) and BMD-MF (%BMD-MF=29.6%,24.5%, respectively), but weak stromal induction without recruitment of BMD-VE or -MF was observed in the tumors of the liver, spleen and lung (%St=9.7%,9.1%,5.4%, respectively). These results indicate that recruitment of BMD-VE and -MF is required for stromal formation during cancer progression, and that the cancer microenvironment is important in the recruitment of BMD-VE and -MF. Less