摘要
目的: 研究茶叶提取物EGCG对人乳腺癌细胞株MDA-MB-231侵袭转移能力的影响及其可能机制。
方法:以高转移人乳腺癌细胞株MDA-MB-231为研究对象,应用糖基化抑制剂苯甲基-α-N-乙酰半乳糖胺为去糖基化阳性对照,以RPMI-1640为阴性对照。MTT法检测EGCG对肿瘤细胞增殖的抑制作用,求出IC50。免疫细胞化学染色法检测MUC1的表达,MTT法检测EGCG对乳腺癌细胞黏附Matrigel能力的影响,Transwell小室进行人工重组基底膜侵袭实验和运动实验,明胶酶谱法(SDS-PAGE)检测肿瘤细胞用药前后分泌Ⅳ型胶原酶MMP-2、MMP-9的差异。
结果:EGCG对人乳腺癌细胞株MDA-MB-231的细胞增殖抑制作用随药物浓度增大和时间延长而增强,其IC50为72.45μg/ml ,用其半量(36μg/ml)作为实验用药浓度。EGCG实验组与未处理组相比,MUC1表达降低,阳性率分别为64%和100%,具有显著性差异(P<0.01),与苯甲基-α-N-乙酰半乳糖胺组的MUC1抑制效应一致。EGCG实验组肿瘤细胞对基底膜的黏附能力下降(P<0.01),未处理组、
苯甲基处理组和EGCG处理组侵袭迁移能力分别为86.5±14.9、64.7±1.2、40.9±10.8和109.0±8.30、76.2±10.69、50.8±5.30。其分泌Ⅳ型胶原酶MMP-2、MMP-9的能力亦明显降低。
结论: 体外实验中,EGCG对MDA-MB-231细胞株的增殖有抑制作用,与时间和剂量呈依赖性;EGCG具有抑制人乳腺癌细胞黏附、侵袭、迁移能力,抑制IV型胶原酶MMP-2、MMP-9的分泌,其机理可能是通过去糖基化干预MUC1的生成。
Objective: To study the effects of tea extract EGCG on invasion and metastasis of breast cancer line MDA-MB-231 and the possible mechanisms.
Methods: Our study was conducted on a highly metastatic human breast cancer cell line MDA-MB-231. Taking benzyl-α-GalNAc(a glycosilation inhinbitor) and RPMI-1640 group as positive and negative control respectively. MTT is performed to determine the growth inhibiting effect of EGCG for getting its IC50. Expression of MUCI was observed by immunocytochemistry. Adhesion of MDA-MB-231 to Matrigel was measured with MTT assay. Invasion and metastatic potential were evaluated with transwell chamber. Collagenase-Ⅳ secretion was dectected
with SDS-PAGE before and after the administration of EGCG .
Results: Our study demonstrated that EGCG had a time/dose- dependent growth inhibiting effect on MDA-MB-231 and the IC50 was 72.45μg/ml. Compared with untreated group, MDA-MB-231 treated with 36μg/ml EGCG displayed decreased MUCI expression (64% vs 100%), while MUC1 expression was not significantly different between benzyl-α-GalNAc group and EGCG group. Compared with untreated group,Adhesion is remarkably decreased. Invasive and metastatic abilities of three groups are 85.5±14.9、64.7±1.2、40.9±10.8 and 109.0±8.30、76.2±10.69、50.8±5.30 respectively,the Collagenase-Ⅳ secretion of MDA-MB-231 cells was also decreased.
Conclusion: In vitro, EGCG has a time/dose-dependent growth inhibiting effect on MDA-MB-231 cells. EGCG may suppress adhesion , metastasis, Invasion, and Collagenase-Ⅳsecretion of MDA-MB-231 cells by inhibiting the production of MUC1 .
引文
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