目的：观察宫颈癌HCE1多细胞球体中整合素β1的表达与其浸润内皮细胞的关系，研究抗整合素β1单克隆抗体P5D2对HCE1多细胞球体浸润人脐静脉内皮细胞的抑制作用。方法：建立宫颈鳞癌细胞HCE1多细胞球体浸润单层人脐静脉内皮细胞HUVEC模型(简称为HCE1浸润模型)。倒置显微镜下观察P5D2干预致HCE1浸润模型的形态学改变,并用Motic Med医学图像软件计算浸润面积的改变。免疫细胞化学SABC法检测HCE1单层细胞、HCE1多细胞球体、HUVEC单层细胞以及P5D2干预前后HCE1浸润模型中整合素β1的表达。结果：成功建立HCE1浸润模型，随着培养时间延长肿瘤多细胞球体生长迅速，培养7 d后HCE1浸润模型中浸润面积是实验开始时的40.42倍。P5D2干预1、4、7 d后的HCE1浸润模型浸润面积明显小于对照组（P<0.05或P<0.01），第7天的浸润面积较对照组减少(8468±0.08)%。HCE1多细胞球体中整合素β1阳性高表达率高于HCE1单层细胞（P<0.01），HUVEC细胞中整合素β1阴性表达； P5D2干预后HCE1浸润模型中整合素β1的高表达率明显低于对照组（P<001）。结论：整合素β1在HCE1多细胞球体及其浸润模型中表达的上调可能与细胞黏附力、侵袭力增强有关；抗整合素β1单克隆抗体P5D2能够部分阻断HCE1 多细胞球体对HUVEC单层细胞的浸润。

Objective:To probe into the relationship between integrin β1 expression in human cervical carcinoma HCE1 multicellular spheroids (HCE1/MCS) and their invasion into human umbilical vein endothelium cells (HUVEC), so as to assess the inhibitory effect of antiintegrin β1 monoclonal antibody P5D2 on the invasion of HCE1/MCS into HUVEC. Methods:A model of HUVEC monolayer invaded by HCE1/MCS was established (in brief, the HCE1 invading model). Morphology changes of the HCE1 invading model were observed under inverted microscope and analyzed by Motic Med System after P5D2 treatment. The expressions of β1 integrin on HCE1 monolayer cells, HCE1/MCS, HVUEC monolayer cells, and P5D2treated HCE1 invading models were measured by immunocytochemistry SABC assay. Results:A HCE1 invading model was successfully established. The HCE1/MCS proliferated rapidly after culture, and on the 7th day the invading area of HCE1/MCS was 40.42 folds larger than the original HCE1/MCS. Invading areas of P5D2treated HCE1/MCS were significantly smaller than that of the control group after 1, 4, 7 day (P<0.05, or P<0.01), with the invading area after 7 days reduced by (84.68±0.08) % compared with the control group. Integrin β1 expression in HCE1/MCS was significantly higher than that in HCE1 monolayer cells (P<0.01), and the expression was negative in HUVEC. Integrin β1 expression in P5D2treated HCE1/MCS was significantly lower than that the in untreated HCE1/MCS (P<0.01). Conclusion:Upregulated expression of integrin β1 in HCE1/MCS and HCE1 invading model may be associated with their enhanced adhesion and invasion abilities. Antiintegrin β1 monoclonal antibody P5D2 can partially block the invasion of HCE1/MCS into HUVEC.

湖南省科技厅科技发展计划重点项目（No.2007SK3043）