Ⅳ型胶原酶对胶质瘤增殖、迁移及侵袭的影响

Effects of type IV collagenase on proliferation, migration and invasion of glioma

目的观察Ⅳ型胶原酶在人脑胶质瘤组织中的表达及其对C6胶质瘤细胞增殖、迁移及侵袭的作用。方法利用Westernblot法比较人脑胶质瘤组织和正常组织中Ⅳ型胶原酶的表达量及含量，将体外培养的C6胶质瘤细胞分为3组，分别加入正常培养液（对照组）、IV型胶原酶（实验组）及Ⅳ型胶原酶＋抑制剂GM6001（抑制组），采用噻唑蓝（MTT）实验、细胞迁移实验和细胞侵袭实验，观察Ⅳ型胶原酶对X6胶质瘤细胞增殖、迁移及侵袭的作用。结果胶质瘤组织中Ⅳ型胶原酶的含量（87．5％）显著高于正常脑组织（20．0％）。吸光度比值半定量分析结果两组间差异也有统计学意义（0．81±0．12比0．04±0．01，P〈0．01）。MTY实验显示实验组、对照组和抑制组3组细胞各个时间点吸光度差异无统计学意义（P〉0．05），3组迁移细胞数目分别为（137．49±14．53）、（65．35±6．57）、（67．42±7．92）个，实验组同对照组和抑制组差异均有统计学意义（P〈0．05），3组侵袭细胞数目分别为（33．67±7．47）、（20．45±4．89）、（21．67±5．83）个，实验组同对照组和抑制组差异均有统计学意义（P〈0．05）。结论Ⅳ型胶原酶在人脑胶质瘤组织中表达率及含量明显增高，对于体外培养的大鼠C6胶质瘤细胞，Ⅳ型胶原酶可显著促进其迁移和侵袭，并通过添加抑制剂GM6001后可有效逆转其迁移和侵袭趋势。

Objective To study the expression of type IV collagenase in human glioma tissue, and the effects of type IV collagenase on proliferation, migration and invasion of Glioma cells, so as to provide basis for clinical treatment. Methods Western blotting was used to compare the content of type IV colla- genase between malignant Glioma tissues and normal tissues. Glioma cell C6 were divided into three groups, respectively added with cell culture medium （control group）, type IV collagenase （study group）, type IV collagenase and specific inhibitor （inhibitor group）. Use the methyl thiazol tetrazolium （MTr） as- say, transwell migration and invasion assay to observe the effects of type IV collagenase on proliferation, migration and invasion of Glioma cells. Results The expression rate of type IV collagenase in malignant glioma tissues （87.5%） is higher than it in normal tissues （20. 0% ）. There was significant difference in relative absorption ratio between two groups （0. 81 ±0. 12 vs. 0.04 ±0. 01 ,P 〈0.01）. The result of MTT assay indicate the cell proliferation ability of the three groups have no statistical significance at every point. The cell count in migration assary of study group （ 137.49 ± 14. 53 ） was significantly more than control group （65.35 ± 6. 57, P 〈 0. 01 ） and inhibitor group （ 67.42 ± 7.92, P 〈 0.01 ）. Similarly, the cell count in invasion assary of study group （ 33.67 ± 7.47 ） was significantly more than control group （20.45±4. 89, P〈0.01） and inhibitor group （21.67 ±5.83, P〈0.01）Conclusion The content and the expression rate of type IV collagenase in Malignant Glioma tissues is higher than it in normal tissues. Type IV collagenase promotes cell migration and invasion, and could be blocked by specific inhibitor, pro- viding new ways in the treatment of malignant glioma.