【摘要】 目的 :锌α2糖脂蛋白(zinc-alpha-2-glycoprotein 1,ZAG)是新近发现的具有复杂功能的蛋白,ZAG参与受精、脂代谢及免疫调节等多种细胞生物学行为,本研究探讨ZAG在大肠癌演进过程中的作用。方法:本研究构建ZAG干扰质粒,并转染入HT-29结肠癌细胞株,免疫印迹与逆转录聚合酶链式反应(RT-PCR)方法检测转染前后ZAG表达的变化;检测转染前后细胞三磷酸腺苷(adenosine triphosphate,ATP)含量的变化,以了解ZAG对细胞代谢的影响;利用小室实验检测转染前后HT-29细胞侵袭能力的变化。结果:ZAG干扰质粒成功转染入HT-29细胞并筛选出稳定转染细胞株;转染后的HT-29细胞ATP水平上调,且转染后的HT-29侵袭能力增强。结论:研究结果提示ZAG可能通过调节ATP水平增强大肠癌HT-29细胞的侵袭能力,为大肠癌治疗提供了有价值的新靶点。更多还原

【Abstract】 Objective:Zinc-alpha-2-glycoprotein 1(ZAG) is a protein with multiple functions,such as participating in lipometabolism,saccharometabolism,energy metabolism,immunoreaction and fertilization. It is observed whether ZAG involved in the regulation of human colorectal cancer cell line(HT-29) in this study. Methods:ZAG-RNAi plasmids were constructed and transfected into human colorectal cancer HT-29 cells. The expressions of target proteins and gene were detected by Western blot and RT-PCR,respectively. Adenosine triphosphate(ATP) was assessed to investigate the activation of energy metabolism. The capacity of HT-29 cell migration was investigated using the Transwell migration assay. Results:ZAG interference plasmids were successfully transfected into HT-29 cells,and stably transfected cell lines were screened out. ATP level and the migration of ZAG inhibitor-treated cells were increased. Conclusion:The down-expression of ZAG promoted the expression of ATP and up-regulated the malignant phenotype ofHT-29 cells.更多还原