中国肺癌杂志
中國肺癌雜誌
중국폐암잡지
Chinese Journal of Lung Cancer
2015年
11期
668-673
,共6页
张翠翠%李月雅%王晶%李凯
張翠翠%李月雅%王晶%李凱
장취취%리월아%왕정%리개
Anip973%HUVEC%细胞增殖%迁移%成管%CD105%CD31%Annexin V
Anip973%HUVEC%細胞增殖%遷移%成管%CD105%CD31%Annexin V
Anip973%HUVEC%세포증식%천이%성관%CD105%CD31%Annexin V
Anip973%HUVEC%Proliferation%Migration%Tube formation%CD105%CD31%Annexin V
背景与目的肿瘤微血管内皮细胞与正常组织来源的微血管内皮细胞相比，具有对生长因子反应性高、粘附因子表达高等特点，造成肿瘤血管通透性高且新生旺盛，导致肿瘤的快速生长和转移。因此了解肿瘤微环境中内皮细胞发生、形态及功能上的异质性特点，对进行合理的、个性化的、以血管内皮细胞为靶点的抗血管新生治疗有一定的指导作用。本实验旨在研究高转移肺腺癌Anip973细胞培养上清液对人脐静脉内皮细胞（human umbilical vein endothelial cell, HUVEC）表面标记及其生物学行为的影响。方法以含有不同浓度Anip973细胞上清液的培养基（RPMI-1640+10%胎牛血清）培养的HUVEC为实验组、单纯培养基培养的HUVEC为对照组，以CCK-8检测各组细胞增殖率、基质胶成管实验检测成管能力、Transwell检测迁移能力、流式细胞术检测细胞表面标记CD105、CD31以及凋亡标记Annexin V的表达，并分析细胞表面标记的表达与生物学行为之间的关系。结果与对照组相比，经Anip973细胞上清液培养的HUVEC增殖更多、且在浓度为250μL/mL作用24 h最明显（P=0.002）；成管及迁移能力亦均较对照组增强，分别在浓度为125μL/mL （P<0.001）、250μL/mL（P=0.002）时达最强；细胞表面标记CD105表达阳性率升高、浓度为250μL/mL （P=0.028）最明显；CD31表达阳性率呈浓度依赖性升高；凋亡细胞比例降低。相关性分析显示：CD105的表达阳性率与细胞增殖及迁移能力呈正相关关系、CD31的表达与生物学行为并无明显相关性。结论 Anip973细胞上清液能促进HUVEC细胞增殖、迁移、血管形成，CD105也随之变化、并可在一定程度上反应其生物学行为。
揹景與目的腫瘤微血管內皮細胞與正常組織來源的微血管內皮細胞相比，具有對生長因子反應性高、粘附因子錶達高等特點，造成腫瘤血管通透性高且新生旺盛，導緻腫瘤的快速生長和轉移。因此瞭解腫瘤微環境中內皮細胞髮生、形態及功能上的異質性特點，對進行閤理的、箇性化的、以血管內皮細胞為靶點的抗血管新生治療有一定的指導作用。本實驗旨在研究高轉移肺腺癌Anip973細胞培養上清液對人臍靜脈內皮細胞（human umbilical vein endothelial cell, HUVEC）錶麵標記及其生物學行為的影響。方法以含有不同濃度Anip973細胞上清液的培養基（RPMI-1640+10%胎牛血清）培養的HUVEC為實驗組、單純培養基培養的HUVEC為對照組，以CCK-8檢測各組細胞增殖率、基質膠成管實驗檢測成管能力、Transwell檢測遷移能力、流式細胞術檢測細胞錶麵標記CD105、CD31以及凋亡標記Annexin V的錶達，併分析細胞錶麵標記的錶達與生物學行為之間的關繫。結果與對照組相比，經Anip973細胞上清液培養的HUVEC增殖更多、且在濃度為250μL/mL作用24 h最明顯（P=0.002）；成管及遷移能力亦均較對照組增彊，分彆在濃度為125μL/mL （P<0.001）、250μL/mL（P=0.002）時達最彊；細胞錶麵標記CD105錶達暘性率升高、濃度為250μL/mL （P=0.028）最明顯；CD31錶達暘性率呈濃度依賴性升高；凋亡細胞比例降低。相關性分析顯示：CD105的錶達暘性率與細胞增殖及遷移能力呈正相關關繫、CD31的錶達與生物學行為併無明顯相關性。結論 Anip973細胞上清液能促進HUVEC細胞增殖、遷移、血管形成，CD105也隨之變化、併可在一定程度上反應其生物學行為。
배경여목적종류미혈관내피세포여정상조직래원적미혈관내피세포상비，구유대생장인자반응성고、점부인자표체고등특점，조성종류혈관통투성고차신생왕성，도치종류적쾌속생장화전이。인차료해종류미배경중내피세포발생、형태급공능상적이질성특점，대진행합리적、개성화적、이혈관내피세포위파점적항혈관신생치료유일정적지도작용。본실험지재연구고전이폐선암Anip973세포배양상청액대인제정맥내피세포（human umbilical vein endothelial cell, HUVEC）표면표기급기생물학행위적영향。방법이함유불동농도Anip973세포상청액적배양기（RPMI-1640+10%태우혈청）배양적HUVEC위실험조、단순배양기배양적HUVEC위대조조，이CCK-8검측각조세포증식솔、기질효성관실험검측성관능력、Transwell검측천이능력、류식세포술검측세포표면표기CD105、CD31이급조망표기Annexin V적표체，병분석세포표면표기적표체여생물학행위지간적관계。결과여대조조상비，경Anip973세포상청액배양적HUVEC증식경다、차재농도위250μL/mL작용24 h최명현（P=0.002）；성관급천이능력역균교대조조증강，분별재농도위125μL/mL （P<0.001）、250μL/mL（P=0.002）시체최강；세포표면표기CD105표체양성솔승고、농도위250μL/mL （P=0.028）최명현；CD31표체양성솔정농도의뢰성승고；조망세포비례강저。상관성분석현시：CD105적표체양성솔여세포증식급천이능력정정상관관계、CD31적표체여생물학행위병무명현상관성。결론 Anip973세포상청액능촉진HUVEC세포증식、천이、혈관형성，CD105야수지변화、병가재일정정도상반응기생물학행위。
Background and objectiveUnlike normal tissue-derived microvascular endothelial cells, tumor mi-crovessel endothelial cells are highly reactive to growth factors and exhibit more adhesion molecules. hTus, vascular tumors are highly permeable and grow vigorously; this occurrence results in rapid growth and metastasis cancer cells. hTerefore, under-standing the characteristics of endothelial cells in the tumor microenvironment guides anti-angiogenic therapy. To this end, we explore the effect of the supernatant obtained from cultured Anip973 cells (high-metastatic human lung adenocarcinoma cells) on the biological behavior and on the cell surface markers of the human umbilical vein endothelial cell (HUVEC).Methods hTe HUVEC that was cultured in a medium (RPMI-1640 + 10% fetal bovine serum) containing various concentrations of Anip973 supernatants was categorized into experimental groups. hTe HUVEC cultured in a medium without Anip973 super-natants served as the control group. Proliferation was determined with CCK-8; blood vessel formation was investigated with three-dimensional culture techniques in vitro; and HUVEC migration was observed via transwell assay. At the same time, the expressions of CD105, CD31, and the apoptotic marker of Annexin V were detected through lfow cytometry for analyzing the relationship between the expression of cell surface markers and biological behavior.Results Following incubation with the supernatant obtained from cultured Anip973 cells, HUVEC proliferated more than the control group did, and the prolifera-tion rate was maximized when incubated in a supernatant concentration of 250 μL/mL for 24 h (P=0.002). In addition, the experimental groups exhibited varying degrees of migration and forms of vascular lumen sample structure, especially at super-natant concentrations of 125 μL/mL (P<0.001) and 250 μL/mL (P=0.002), respectively. CD105 expression was optimized at 250 μL/mL (P=0.028), and CD31 expression also increased with an increase in concentration. However, the percentage of apoptotic cells decreased. Correlation analysis results showed that cell proliferation, migration, and CD105 expression were signiifcantly and positively correlated with one another. By contrast, no signiifcant correlation was detected between CD31 expression and biological behavior.ConclusionAnip973 supernatants can promote HUVEC proliferation and migration, as well as angiogenesis. In addition, cell surface markers can change concurrently and relatively. To a certain extent, changes in CD105 expression can be attributed to shitfs in its biological behavior.
