人肝癌裸鼠皮下移植瘤早期血液供应模式的定量研究

目的 定量评估不同时间点人肝癌裸鼠皮下移植瘤内血液供应模式的变化。方法 将1×10⁷/mL的MHCC97-H细胞悬液同一时间接种于40只裸鼠（0.2 mL/鼠）的左侧腹股沟皮下，分别于种瘤后第2、3、4、5周取瘤行HE染色和免疫组化染色（CD34和CD34/PAS双染色），定量分析血管参数，包括微血管面积（microvascular area，MVA）、微血管密度（microvessel density，MVD）、血管生成拟态（vasculogenic mimicry，VM），评估肿瘤内血管供应模式的变化。结果 随着肿瘤生长时间的延长，MVA先减小后增大，第3周肿瘤MVA小于第2周（13 720.8±2 375.6）μm² vs.（28 795.5±3 412.7）μm²，P=0.013]，第4周肿瘤MVA大于第3周（29 221.3±3 958.5）μm² vs.（13 720.8±2 375.6）μm²，P=0.025]；VM百分比呈明显升高趋势，第4周肿瘤VM百分比高于第2周（84.2%±15.4% vs.50.1%±14.6%，P=0.015）。结论 在人肝癌裸鼠皮下移植瘤新生血管形成的早期阶段，随着肿瘤生长时间的延长，VM逐渐成为肿瘤的主要血液供应模式。

Quantitative study on blood supply patterns in the early stage of human hepatocellular carcinoma subcutaneous xenografts in nude mice

Objective To quantitatively evaluate the changes of blood supply patterns in the early stage of human hepatocellular carcinoma subcutaneous xenografts in nude mice at different time points. Methods 0.2 mL of the MHCC97-H cell suspension, which was at the density of 1×10⁷/mL,was injected into the subcutaneous tissue of 40 nude mice left inguinal region respectively at the same time.The tumors were dissected at the 2nd,3rd,4th and 5th weeks after injection.The paraffin sections were stained with hematoxylin-eosin (HE) and immunohistochemistry method,including CD34,CD34 and acid-Schiff (PAS) dual-staining.The microvascular parameters,which included microvascular area (MVA),microvessel density (MVD),vasculogenic mimicry (VM) parameters,were quantitatively analyzed at different time points of tumor growth to evaluate the changes on blood supply pattern. Results The MVA was lower at the 3rd week than the 2nd(13 720.8±2 375.6) μm² vs.(28 795.5±3 412.7) μm²,P=0.013],and higher at the 4th week than the 3rd week(29 221.3±3 958.5) μm² vs.(13 720.8±2 375.6) μm²,P=0.025].The VM percentage was lower at the 2nd week than the 4th week (50.1%±14.6% vs. 84.2%±15.4%,P=0.015). Conclusion In the early stage of neovascularization in human hepatocellular carcinoma subcutaneous xenografts of nude mice,VM gradually became the main blood supply mode as time went on.