【摘要】 目的:探讨CD73在乳腺癌细胞中的表达特征及其与乳腺癌细胞侵袭转移的相关性。方法:①采用半定量RT-PCR法和高压液相层析法检测6种乳腺癌细胞株CD73的mRNA表达及CD73的活性。②构建CD73真核表达载体pcDNA-NT5E,采用脂质体转染法将其导入无侵袭力、CD73低表达的乳腺癌细胞T47D,同时以空载体质粒pcDNA和未转染细胞作为对照。利用Transwell体外侵袭力实验和运动力实验,观察转染pcDNA-NT5E对T47D细胞侵袭力和运动力的影响。结果:①高度恶性乳腺癌细胞的CD73mRNA水平和CD73活性均显著高于低度恶性细胞(P<0·05)。②转染后细胞T47D的CD73基因转录、CD73酶活性、细胞的侵袭力和运动力明显高于转染前。结论:乳腺癌细胞的恶性程度与CD73表达呈正相关。CD73可增强人乳腺癌细胞的侵袭力和运动力。提示在乳腺癌转移过程中CD73发挥重要的促进作用。更多还原

【Abstract】 AIM: To understand the expression of ecto-5’-nucleotidase (CD73) and explore its effect on the motility and invasion ability of breast cancer cell lines. METHODS: ① CD73 mRNA expression was analyzed by semiquantitative RT-PCR. Surface CD73 activity in breast carcinoma cell lines was measured by HPLC according to the conversion of E-AMP to E-ADO. ② Plasmid pcDNA-NT5E was transfected into human low metastatic breast cancer cell line T47D with lipofectinamine2000. CD73 mRNA expression was detected by semiquantitative RT-PCR and surface CD73 activity was determined by HPLC before and after transfection. The effect of pcDNA-NT5E transfection on motility and invasion ability was observed by in vitro invasion assay. RESULTS: ① The RNA expression and enzyme activity of CD73 were variable in different breast cancer cell lines. ② The transfect-T47D overexpressed CD73 at mRNA level and with high enzyme activity. Constitutively active CD73 drove cells transmembrane and invasion into matrigel. Promotion effects were statistically significant (P<0.05) between pcDNA-NT5E-T47D cell and negative control. CONCLUSION: The CD73 mRNA expression and its enzyme activity in a panel of breast cancer cell lines differ in invasive and metastatic potential. CD73 increases the motility and invasion abilities sharply in the pcDNA-NT5E-T47D cells.更多还原