摘要
目的明确不同微环境对肿瘤细胞侵袭能力和微循环模式的影响,并阐述其相关分子机制。方法将恶性黑色素瘤单细胞悬液 B16分别接种至 C57小鼠腹腔和后肢肌肉组织中,于接种肿瘤后第18天后处死动物,取腹腔内肿瘤组织和小鼠后肢肿瘤组织,免疫组织化学(SP 法)CK18染色对后肢组和腹腔组肿瘤细胞恶性侵袭表型进行比较,并比较在不同微环境生长下肿瘤细胞缺氧诱导因子(HIF)-1α的表达,免疫组织化学染色和 Real time PCR 方法从蛋白水平和 mRNA 水平比较腹腔组和后肢组肿瘤细胞基质金属蛋白酶(MMP)-2和 MMP-9的表达差别;并对肿瘤组织内不同微循环模式进行计数。结果免疫组织化学染色结果显示与腹腔组相比,后肢组肿瘤细胞 CK18、HIF-1α表达明显增高(t=8.142,t=3.645,P 均<0.05),同时也表达更高的肿瘤侵袭相关蛋白,肿瘤细胞表达 MMP-2和 MMP-9均明显高于腹腔组(t=4.916,t=7.782,P 均<0.05),Real time PCR 检测结果也显示后肢组 MMP-2和 MMP-9 mRNA 表达也明显高于腹腔组(t=36.814,t=26.025,P 均<0.05)。在后肢组肿瘤组织中,血管生成拟态是肿瘤细胞获得营养的主要方式,而腹腔组肿瘤细胞获得营养供应方式主要通过内皮依赖性血管。结论不同生长微环境影响黑色素瘤细胞的恶性表型转化,致使肿瘤细胞分泌更多肿瘤侵袭转移相关蛋白,促进肿瘤细胞向周围组织浸润,肿瘤细胞侵袭能力的增高促进了肿瘤血管生成拟态的形成。
Objective To investigate the influence of different microenvironments on tumor microcirculation patterns and invasive capability. Methods Melanoma B16 cells were injected into the peritoneal cavity and skeletal muscle of C57 mice synchronously. CK18 expression in melanoma was assessed to distinguish the malignant phenotype of tumors in the peritoneal cavity from that in the skeletal muscle. HIF-lot, MMP-2 and MMP-9 protein and mRNA expression were compared in the two microenvironments. Cells positive for each immunohistochemical stain and the vessels representative of each type of microcirculation pattern were evaluated in two microenvironments. Results CK18 and HIF-1α expression in melanoma were significantly higher in the skeletal muscle than in the peritoneal cavity （t = 8. 142 ,t = 3. 645, P 〈 0. 05 ）. Compared with the peritoneal cavity, melanoma cells in the skeletal muscle overexpressed MMP- 2 and MMP-9（t =4. 916, t =7. 782, P〈0. 05）. Real time-PCR results also showed that MMP-2 and MMP- 9 mRNA levels in melanoma were higher in the skeletal muscle than in the peritoneal cavity （t = 36. 814 ,t = 26. 025,P 〈 0. 05 ）. Vasculogenic mimicry channels and endothelium-dependent vessels were the major microcirculation patterns in the skeletal muscle and in the peritoneal cavity respectively. Conclusions Different microenvironments affect invasiveness and blood supply patterns of melanoma. Different microenvironment induced tumor cell secretion of more invasion-related proteins and affect invasiveness and blood supply patterns of melanoma.
出处
《中华病理学杂志》
CAS
CSCD
北大核心
2007年第12期832-837,共6页
Chinese Journal of Pathology
基金
国家自然科学基金(30370554)
