Abstract:

Objective: To explore the potential of BB-94, a matrix metalloproteinase-9 (MMP-9) inhibitor, in alleviating malignant biological behavior of human oral squamous cell carcinoma (OSCC) cell line CAL27 and its possible mechanism. Methods: CAL27 cells in logarithmic growth phase were cultured in BB-94 culture medium with different concentrations (0, 0.125, 0.25, 0.5, 1.0 μg/mL) for 48 hours; the activity of MMP-9 in cell culture supernatant was analyzed by gelatin zymogram; the cell survival rate was detected by cell counting kit-8 (CCK-8) method; the apoptosis, migration, and invasion of CAL27 cells were detected by flow cytometry, wound healing test and Transwell test; real-time quantitative polymerase chain reaction (RT-qPCR) was used to detect the mRNA expression levels of phosphatidylinositol 3-kinase (PI3K) and protein kinase B (Akt); the expressions of PI3K, phosphorylated protein kinase B (p-Akt), MMP-9, vimentin (Vim), N-cadherin (N-cad) and E-cadherin (E-cad) proteins were detected by Western blotting. Results: The relative activity of MMP-9, the survival rate of CAL27 cells, the wound healing rate, the number of cell invasion, the relative expression level of PI3K and Akt mRNA and the relative expression level of PI3K, p-Akt, Vim, N-cad and MMP-9 protein decreased with the increase in the concentration of BB-94 (P<0.05). The apoptosis rate of CAL27 cells and the relative expression level of E-cad protein increased as the concentration of BB-94 went up (P<0.05). Conclusion: These results indicate that MMP-9 inhibitor could inhibit the malignant biological behavior of human OSCC cell line CAL27, and its mechanism may be related to preventing the activation of PI3K/Akt signaling pathway and the progress of epithelial mesenchymal transition (EMT).

Key words: matrix metalloproteinase-9 inhibitor, human oral squamous cell carcinoma, malignant biological behavior, phosphatidylinositol 3-kinase, protein kinase B

摘要：

目的：探究基质金属蛋白酶-9（matrix metalloproteinase-9，MMP-9）抑制剂（BB-94）对人口腔鳞状细胞癌（oral squamous cell carcinoma，OSCC）细胞系CAL27恶性生物学行为的影响及可能的作用机制。方法：用含不同浓度（0、0.125、0.25、0.5、1.0 μg/mL）BB-94培养液培养对数生长期CAL27细胞48 h，明胶酶谱实验分析细胞培养上清液中MMP-9的活性；细胞计数试剂盒-8（cell counting kit‐8，CCK-8）实验检测细胞存活率；流式细胞术、伤口愈合实验及Transwell实验检测CAL27细胞凋亡、迁移及侵袭情况；实时定量聚合酶链反应（real-time quantitative polymerase chain reaction，RT-qPCR）技术检测磷脂酰肌醇3-激酶（phosphatidylinositol 3-kinase，PI3K）、蛋白激酶B（protein kinase B， PKB，又称Akt）mRNA表达水平；蛋白质印迹法（Western blotting）检测PI3K、磷酸化蛋白激酶B（phosphorylated protein kinase B，又称p-Akt）、MMP-9、波形蛋白 （vimentin，Vim）、神经钙黏素 （N-cadherin，N-cad）、上皮钙黏素 （E-cadherin，E-cad）表达情况。结果：CAL27细胞MMP-9相对活性、存活率、划痕愈合率、细胞侵袭数、PI3K和Akt mRNA相对表达水平及PI3K、p-Akt、Vim、N-cad、MMP-9蛋白相对表达水平均随BB-94作用浓度增大而降低（P<0.05），CAL27细胞凋亡率、E-cad蛋白相对表达水平均随BB-94作用浓度增大而升高（P<0.05）。结论：MMP-9抑制剂可抑制人口腔鳞状细胞癌细胞系CAL27恶性生物学行为，其作用机制可能与阻止PI3K/Akt信号通路激活及上皮间质转化（epithelial mesenchymal transition，EMT）进展有关。

关键词: 基质金属蛋白酶-9抑制剂, 人口腔鳞状细胞癌, 恶性生物学行为, 磷脂酰肌醇3-激酶, 蛋白激酶B

CLC Number: 

• R739.5