微环境中肿瘤新生血管巨噬细胞和α平滑肌动蛋白的表达及其与肝癌患者预后的关系

The prognostic analysis of hepatocellular carcinoma based on the tumor neo-vessels, macrophages and a-SMA in tumor microenvironment

目的探讨肝癌微环境中肿瘤新生血管、肿瘤相关巨噬细胞和成纤维细胞α平滑肌动蛋白（α－SMA）的表达及其与肝癌患者预后的关系。方法收集101例肝癌患者的临床病理资料，采用免疫组化法检测肝癌组织新生血管、巨噬细胞和α－SMA的表达，采用配备有Olympus－DP72图像采集系统和CRi Nuance多光谱成像系统的Olympus－BX51正置显微镜进行拍照和定量分析。结果101例肝癌患者的中位无进展生存时间（DFS）为5个月，其中98例患者的肿瘤新生血管数量为51-429个，中位值为218个；91例患者的巨噬细胞数量为110-555个，中位值为259个；100例患者的肿瘤间质厚度为35.61-555.35 μm，中位值为246.98 μm。肿瘤新生血管低密度组和高密度组患者的中位DFS分别为7和5个月，差异无统计学意义（P＝0.197）；巨噬细胞低密度组和高密度组患者的中位DFS分别为8和4个月，差异有统计学意义（P＝0.022）；肝癌间质成纤维细胞低密度组和高密度组患者的中位DFS分别为12和3个月，差异有统计学意义（P〈0.001）。联合低表达组、联合中表达组和联合高表达组（新生血管、巨噬细胞和成纤维细胞3种间质成分均呈高表达为联合高表达组，3种间质成分均呈低表达为联合低表达组，其余为联合中表达组）患者的中位DFS分别为17、5和3个月，差异有统计学意义（P＝0.001）。Cox回归分析结果显示，肝硬化、肿瘤复发、巨噬细胞密度和成纤维细胞密度均为肝癌患者的独立预后因素（均P〈0.05）。结论肝癌间质微环境中肿瘤新生血管、巨噬细胞和α－SMA存在异质性，对肝癌的侵袭转移有重要作用。

Objective To analyze the quantitative expression and prognostic significance of tumor neo-vessels, macrophages and fibroblasts in tumor microenvironment of hepatocellular carcinoma （HCC）. Methods The clinic-pathological features and tissue samples for 101 HCC cases were collected. Immunohistochemistry was used to stain the tumor neo-vessels, macrophages and fibroblasts on tumor tissue. The distribution results and quantitative data of above key components were acquired by inverted microscopy equipped with CRi Nuance multispectral analysis system. The number of tumor neo-vessels and macrophages on HCC tissue were counted and the thickness of cancer stroma based on the expression of fibroblasts was measured. The clinic-pathological characteristics and overall survival were analyzed. Results The median disease free survival （DFS） of 101 HCC cases was 5 month. The quantitative analysis of tumor neo-vessels,macrophages and fibroblasts showed that the expression range was 51-429 with median 218, 110-555 with median 259, 35.61-555.35 with median 246.98, respectively. To take the median as cutoff, all the cases could be classified into high and low expression group. The survival analysis demonstrated that the high density group of maerophages （P = 0.022） and fibroblasts （P〈 0.001 ） has shorter DFS than low density group, with statistical significance. The high tumor neo-vessels group has shorter DFS with median 5 month than low density group with median 7 month. However, there was no statistical significance between these two group （ P = 0.19＂7 ）. Combined with above the three stromal components, all the cases could be classified into low, middle and high group. The survival analysis demonstrated that the high density group of stromal components has shorter DFS than the other two groups with median 3 month （ P = 0.001 ）. Multivariate analysis by Cox regression indicated that cirrhosis, metastasis status, macrophages and fibroblasts density were the independent prognostic factors. Conclusion The key elements in tumor mieroenvironment including tumor neo-vessels, macrophages and fibroblasts were heterogenic in HCC tissues and played significant roles in HCC invasion and metastasis.