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

众所周知，癌间质相互作用在癌症进展过程中起着重要作用。将人胰腺癌细胞系Capan-1皮下异种移植到SCID受者体内，并在第14天和第28天分析基质形成情况。免疫组织化学和免疫荧光研究显示，肿瘤巢内和周围存在bm来源的内皮细胞（X-gal/CD31或H-2b/CD31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）。第14天，脑源性内皮细胞（BMD-VE）和脑源性肌成纤维细胞（BMD-MF）的频率分别为25.3±4.4%和12.7±9.6%。第28天，bm来源的内皮细胞的频率为26.7±9.7%，与第14天的值相似。而第28天bm源性肌成纤维细胞的频率（39.8±17.1%）明显高于第14天（P<0.05）。h -2b阳性肌成纤维细胞的拓扑异构酶α α阳性率为2.2±1.2%，而h -2b阴性肌成纤维细胞的拓扑异构酶α α阳性率仅为0.3±0.4%，bm源性肌成纤维细胞的增殖活性显著（P<0.05）。我们的研究结果表明，bm来源的肌成纤维细胞在肿瘤发展的后期成为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差异对BMD-VE和BMD-MF募集的影响，我们将10种人癌细胞系植入双突变小鼠骨髓移植的免疫缺陷小鼠皮下组织（rag1 -/- b-gal Tg或rag1 -/- GFP Tg）。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）和基质比例（%St）。分析了两者之间的相关性。尽管%BMD-VE和%BMD-MF因癌细胞系而异（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）。%BMD-VE与%BMD-MF也有显著相关（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个不同的部位：皮下组织、腹膜、肝脏、脾脏和肺。肿瘤在皮下组织和腹膜中诱导间质增生（%St分别为22.7%、19.5%），含有BMD-VE （%BMD-VE分别为21.6%、16.5%）和BMD-MF (%BMD-MF分别为29.6%、24.5%)，但在肝脏、脾脏和肺部肿瘤中，间质诱导较弱，未募集BMD-VE或-MF （%St分别为9.7%、9.1%、5.4%）。这些结果表明，在癌症进展过程中，基质的形成需要BMD-VE和-MF的募集，而癌症微环境在BMD-VE和-MF的募集中很重要。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