间质液压与涎腺腺样囊性癌恶性表型的关系及相关分子机制

摘要

Objective To explore the effects of stress imposed on adenoid cystic carcinoma (ACC), therefore to clarify the molecular basis and mechanism of ACC’s malignant phenotype under the elevated tumor interstitial fluid pressure. Methods ACC cells were cultured under pressure (103.74 kPa), and were divided into four groups (3 h group, 6 h group, 12 h group, 24 h group) according the pressure time. Untreated ACC2 was as negative control group, untreated ACCM was as positive control group. The level of epidermal growth factor receptor (EGFR) was detected by semiquantitative analysis of immunoche-mistry. Matrix metalloproteinase 9 (MMP9) and EGFR mRNA expression were assessed by reverse transcriptase polymerase chain reaction. EGFR, phosphorylation epidermal growth factor receptor (P-EGFR), MMP9, keratinocyte growth factor (KGF) and phosphorylation extracellular signal-regulated kinase (P-ERK) protein expressions were assessed by Western blot. Results As the extension of pressure time, the expression of EGFR, P-EGFR, MMP9, KGF, P-ERK in ACC2 gradually increased, which were positive correlation with pressure time, and were higher than that of negative control group. Conclusion Under the stimulation of pressure, the mRNA and protein levels of adhesion molecules and metastatic relative molecules in ACC2 were sharply elevated.%目的：探讨腺样囊性癌（ACC）细胞受力后的基因表达及调控物质变化特点，从而阐明间质液压促进ACC细胞恶性表型获得的分子基础及其作用的分子机制。方法加压（103.74kPa）培养ACC2细胞，根据加压时间不同将实验组分为3h组、6h组、12h组、24h组。以未加压培养的ACC2为阴性对照，未加压培养的的ACCM为阳性对照。采用免疫组织化学法半定量检测各组表皮生长因子受体（EGFR）的表达，逆转录聚合酶链反应法检测基质金属蛋白酶（MMP）9和EGFRmRNA表达，Westernblot法检测MMP9、EGFR、磷酸化表皮生长因子受体（P-EGFR）、角质形成细胞生长因子（KGF）、磷酸化细胞外信号调节激酶（P-ERK）蛋白表达。结果随着加压时间的延长，ACC2细胞内EGFR、P-EGFR、MMP-9、KGF、P-ERK的表达逐渐增加，整体呈时间正相关，且均高于阴性对照组。结论在力学刺激下，ACC2细胞中黏附、转移相关分子的mRNA及蛋白表达水平均升高。