CAJ | 학술논문

目的研究不同微环境对黑色素瘤局部循环模式的影响及其相关分子机制.方法实验研究.将60只C57BL/6J小鼠随机分皮下组和眼内组,每组30只,将B16细胞悬液分别接种到鼠鼷部皮下与视网膜下腔,接种后14 d取材,检测移植瘤扩散与转移情况及瘤组织局部微循环模式,计数并比较血管生成拟态(MV)、马赛克血管(VM)、内皮依赖性血管(E);检查肿瘤组织的扩散及转移情况.采用实时聚合酶链反应(RT-PCR)及免疫组化技术检测酪氨酸激酶受体-2(EphA2)、基质金属蛋白酶-2(MMP-2)及基质金属蛋白酶-2(MMP-9)蛋白及其mRNA表达.进一步分析移植瘤中VM与EphA2、MMP-2及MMP-9 mRNA表达量的相关性.利用两组独立样本t检验分析眼内组与皮下组肿瘤组织各指标的差别,组内各指标之间的相关性用Pearson方法分析.结果黑色素瘤眼内与皮下移植瘤中3种微循环模式共存,并且眼内组VM较皮下组增多,两组比较具有统计学意义(t=4.188,P=0.000),而MV和E在皮下移植瘤中高于眼内移植瘤,但两组比较无统计学差异(t=1.473,1.805;P=0.146,0.076).眼内组EphA2、MMP-2和MMP-9 mRNA表达量显著增加,高于皮下组(t=3.642,8.109,9.357.;P=0.002,0.001,0.001).在EphA2表达增高的同时VM也相应增多,二者呈正相关(r=0.412,P=0.021);VM增多与MMP-2和MMP-9表达增加也存在一定关系,但差异无统计学意义(P>0.05).眼内移植瘤中发现眶内扩散5例,肺转移6例.结论微环境变化可使黑色素瘤的微循环模式发生改变,眼内移植瘤细胞可通过增加EphA2、MMP-2和MMP-9表达使VM形成增加,使VM成为眼内移植瘤主要供血模式,从而使肿瘤更具有侵袭性.(中华眼科杂志,2009,45:641-646)
목적 연구불동미배경대흑색소류국부순배모식적영향급기상관분자궤제.방법 실험연구.장60지C57BL/6J소서수궤분피하조화안내조,매조30지,장B16세포현액분별접충도서혜부피하여시망막하강,접충후14 d취재,검측이식류확산여전이정황급류조직국부미순배모식,계수병비교혈관생성의태(MV)、마새극혈관(VM)、내피의뢰성혈관(E);검사종류조직적확산급전이정황.채용실시취합매련반응(RT-PCR)급면역조화기술검측락안산격매수체-2(EphA2)、기질금속단백매-2(MMP-2)급기질금속단백매-2(MMP-9)단백급기mRNA표체.진일보분석이식류중VM여EphA2、MMP-2급MMP-9 mRNA표체량적상관성.이용량조독립양본t검험분석안내조여피하조종류조직각지표적차별,조내각지표지간적상관성용Pearson방법분석.결과 흑색소류안내여피하이식류중3충미순배모식공존,병차안내조VM교피하조증다,량조비교구유통계학의의(t=4.188,P=0.000),이MV화E재피하이식류중고우안내이식류,단량조비교무통계학차이(t=1.473,1.805;P=0.146,0.076).안내조EphA2、MMP-2화MMP-9 mRNA표체량현저증가,고우피하조(t=3.642,8.109,9.357.;P=0.002,0.001,0.001).재EphA2표체증고적동시VM야상응증다,이자정정상관(r=0.412,P=0.021);VM증다여MMP-2화MMP-9표체증가야존재일정관계,단차이무통계학의의(P>0.05).안내이식류중발현광내확산5례,폐전이6례.결론 미배경변화가사흑색소류적미순배모식발생개변,안내이식류세포가통과증가EphA2、MMP-2화MMP-9표체사VM형성증가,사VM성위안내이식류주요공혈모식,종이사종류경구유침습성.(중화안과잡지,2009,45:641-646)
Objective To investigate the influence of different micreenvironments on melanoma vasculogenic mimicry, invasiveness and metastasis behavior. Methods It was an experimental study. Sixty C57BL/6J mice were randomly divided into two groups with 30 mice per group. Melanoma B16 cells were injected into the subretinai space and groin area of mice synchronously. The number of each type of microcireulation pattern was counted. The invasion and metastasis were observed. EphA2, MMP-2 and MMP-9 expression and their mRNA levels were detected by immunohistochemical staining and real time RT-PCR and compared between two groups. Results Five invasions and six lung metastases were found in the subretinal group while no invasion and metastasis were found in the groin group. The number of VM channels was significantly higher in subretinal group (t =4. 188, P =0. 000). However, no significant difference of mosaic vessel and endothelium-dependent vessel was observed between two groups (t = 1. 473, 1. 805 ; P = 0. 146, 0.076, respectively). EphA2, MMP-2 and MMP-9 expression was significantly higher in the subretinal group (data not shown). The mRNA levels of EphA2, MMP-2 and MMP-9 were rather higher in the subretinal tumor (t =3.642, 8. 109,9.357;P =0.002, 0.001 and 0.001, respectively). There was a positive association in melanoma cells of the VM between expression of EphA2 (r = 0. 412, P = 0. 021) but no statistically significant correlation between VM and MMP-2 (P > 0. 05), nor between VM and MMP-9. Conclusions Different microenvironments affect invasiveness and blood supply patterns of melanoma. Melanoma cells in intraocular microenvironment increased EphA2 expression which induced the formation of VM channels. Moreover, the expression of MMP-2 and MMP-9 in tumor tissue increased to enhance the invasiveness and metastasis behavior. (Chin J Ophthalmol , 2009,45:641-646)