乳腺癌MMP-2、MMP-9的体外阻断研究

目的:探讨乳腺癌MMP-2、MMP-9表达的临床病理联系及直接阻断MMP-2、MMP-9对乳腺癌细胞增殖、侵袭及细胞周期的影响。方法:应用免疫组织化学方法检测48例乳腺癌MMP-2、MMP-9的表达。明胶酶谱法检测乳腺癌MCF-7细胞MMP-2、MMP-9的表达。MCF-7细胞接种于预铺BME的Tran-swell小室上室,与MMP-2、MMP-9的阻断剂CTT共孵育16h,观察阻断MMP-2、MMP-9对MCF-7细胞侵袭能力的影响。MCF-7细胞与CTT共孵育12h后,MTT法检测阻断MMP-2、MMP-9对细胞增殖能力的影响。MCF-7细胞与CTT共孵育24h后,PI染色流式法检测阻断MMP-2、MMP-9对细胞凋亡和细胞周期的影响。结果:48例乳腺癌组织有MMP-2表达者45例(93.75%),MMP-9表达者38例(79.17%),MMP-2和MMP-9在发生淋巴结转移组明显高表达(P<0.05);组织学分级越高,MMP-2和MMP-9的表达越显著(P<0.05)。明胶酶谱法检测:在MCF-7培养上清中检测到MMP-2、MMP-9的表达。在浓度为100μg/ml和200μg/ml时CTT对MCF-7细胞的侵袭抑制率达到39.5%和61.9%。在终浓度为50、100、200μg/ml时,CTT对MCF-7细胞的增殖率均无明显下降(P>0.05)。CTT处理组MCF-7细胞经流式法检测未见凋亡峰,细胞周期各期未见明显阻滞(P>0.05)。结论:MMP-2、MMP-9与乳腺癌的淋巴结转移密切相关,阻断MMP-2、MMP-9可有效抑制乳腺癌的浸润和转移。

The study of blocking MMP-2/-9 on breast cancer cells

Objective：To study the clinicoPathological significance of MMP-2/-9 exPressions in breast cancer, and the blocking MMP-2/-9 on breast cancer cells. Methods：The exPressions of MMP-2/-9 were examined in 48 cases of breast cancer by immunohistochemistry. The exPressions of MMP-2/-9 in MCF-7 cells were detected by means of gelatin zymogram. MCF-7 cells were incubated with different concentrations of CTT（MMP-2/-9 blocker）in transwell Precoated with BME for 16h,the invasion of MCF-7 cells was detected. MCF-7 cells were in-cubated with different concentrations of CTT in 96 - microwell Polystyrene Plates for 12h,the Proliferation of MCF-7 cells was detected by means of MTT. Incubated with 200μg/ ml of CTT for 24h,aPoPtosis and cell cycle of MCF-7 cells were detected by means of PI staining flow cytometry. Results：The result of immunohistochemistry：Among 48 breast cancer examPles,there were 45 cases exPressing MMP - 2（93. 75% ）and 38 cases exPressing MMP - 9 （79. 17% ）. The exPressions of MMP-2/-9 in breast cancer with lymPh node metastasis grouP was higher than no metastasis grouP（P ﹤ 0. 05）. The result of gelatin zymogram：MMP-2/-9 could be detected in MCF-7 cell cul-ture suPernatant. Cultured with 100μg/ ml and 200μg/ ml of PePtide CTT,the inhibition ratio of cellular invasion of MCF-7 were 39. 5% and 61. 9% . At all concentrations,the Proliferation ratio of MCF-7 had no significant differ-ence between CTT and blank grouP（P ﹥ 0. 05）. The result of flow cytometry showed that no aPoPtotic Peak could be found in MCF-7 cells treated with CTT,and the whole cell cycle had no significant difference between CTT and blank grouP（P ﹥ 0. 05）. Conclusion：In breast cancer,the excessive exPressions of MMP-2/-9 may indicate axil-lary lymPh node metastasis,blocking the function of MMP-2/-9 can inhibit the metastasis of breast cancer.