乌司他丁抑制人乳腺癌细胞的侵袭和转移及其分子机制

The molecular mechanism of ulinastatin as a inhibitor to the invasion and metastasis of human breast cancer cell

目的通过测定不同浓度的乌司他丁(UTI)对体外乳腺癌细胞MCF-7(雌激素受体阳性)和MDA-MB-231(雌激素受体阴性)干预后细胞中乙酰肝素酶(Hpse)、1型透明质酸酶(HYAL1)及细胞黏附分子CD44v6蛋白的表达情况,探讨其对癌细胞侵袭、转移抑制的分子机制。方法将体外培养的MCF-7及MDA-MB-231细胞分别随机分为:(1)对照组;(2)UTI低剂量干预组;(3)中剂量干预组;(4)高剂量干预组;(5)UTI中剂量+泰索帝(TXT)组。蛋白质印迹法(Western blot)检测各组细胞Hpse、HYAL1和CD44v6蛋白的表达;Matrigel穿膜侵袭实验检测细胞侵袭能力;Transwell小室细胞跨膜实验测定细胞的迁移能力。结果癌细胞中Hpse、HYAL1和CD44v6蛋白的表达随UTI浓度增加而降低,UTI与TXT联合用药抑制作用更加显著(P<0.05);乳腺癌MCF-7和MDA-MB-231细胞的侵袭和迁移能力显著下降,UTI与TXT联合处理抑制作用更加明显(P<0.05)。结论 UTI抑制乳腺癌细胞的侵袭和转移可能与Hpse、HYAL1、CD44v6蛋白的表达降低有关。

Objective To observe the effect of ulinastatin at different concentration on the expression of Heparanase（Hpse）, Hyaluronidase-1 and CD44v6 and the invasion and metastasis of human breast cancer cell in vitro, and to explore its molecular mechanism. Methods The breast cancer cell line MCF-7 and MDA-MB-231 cultured in vitro were treated in randomly divided： 1. Control, 2. UTI at the low-dosage group, 3. medium-dosage group, 4. high-dosage group, 5. moderate dosage of UTI with Taxotere （TXT）. The proliferation ability of cells was tested by MTT assay. Western blotting applications were used to measure the expression of Hpse, HYAL1 and CD44v6 ;The invasion ability of the cell lines were tested by Transwell chamber assay with Matrigel;The migration of the two cell lines were examined by Transwell chamber. Results The expression of Hpse, HYAL1 and CD44v6 was significantly decreased compared with the control group,and the effect of that received two medicament simultaneously was superior to that UTI alone（P;0.05） ;compared with the control cells, UTI inhibited the invasion and migration of the breast cancer cell MCF-7 and MDA-MB-231,more significant joint UTI and TXT（P;0.05）. Conclusion UTI can inhibit the invasion and metastasis of the breast cancer cell,the mechanism may be associated with UTI reduced the expression of Hpse, HYAL1 and CD44V6.