MTA1基因对宫颈癌细胞HeLa侵袭转移影响.docVIP

MTA1基因对宫颈癌细胞HeLa侵袭转移影响[摘要] 目的 探索MTA1基因与宫颈癌细胞HeLa侵袭迁移能力的关系。 方法 以脂质体LipofectamineTM2000介导的方法将含有MTA1全长基因的质粒pEGFP-C1/MTA1转染HeLa细胞，应用Western blotting检测转染效果，细胞黏附实验检测细胞黏附能力，细胞划痕实验检测细胞迁移能力的变化，Transwell小室侵袭实验检测细胞侵袭能力的变化。结果 MTA1质粒转染的宫颈癌HeLa细胞中，MTA1蛋白的表达、黏附能力、迁移及侵袭能力均明显高于转染空载体组及未转染组（P＜0.05）。结论 MTA1基因与宫颈癌细胞的侵袭迁移能力有关，MTA1基因可能成为宫颈癌治疗的一个新的靶点。 [关键词] 转移相关基因1；宫颈癌；肿瘤转移 [中图分类号] R737.33[文献标识码] A[文章编号] 1673-9701（2011）20-15-03 The Influence of MTA1 Gene in Invasion and Metastasis of Cervical Cancer ZHU YimingYANG Zhengyan Department of Gynecology Oncology，Zhejiang Cancer Hospital，Hangzhou 310022，China [Abstract] Objective To study the relativity between MTA1 and invasion and migration of cervical cancer cells. Methods The plasmid pEGFP-C1/MTA1 inserted by the full length cDNA encoding human MTA1 gene was transiently transfected into HeLa cells by LipofectamineTM2000. The expression of MTA1 protein was investigated by Western blotting. Cell adhesion ability change was detected by cellular adhesive experiment. Migration rate was measured by wound healing assay. Furthermore，the abilities of cell invasion and metastasis were investigated using Transwell chambers. Results The cell adhesion，invasion and migration ability was increased after MTA1 over-expression mediated by pEGFP-C1/MTA1 transfection in HeLa cells（P＜0.05）. Conclusion Over-expression of MTA1 gene significantly increased the cell invasion，migration，and adhesion ability. MTA1 could serve as a potential therapeutic target in cervical cancer. [Key words] Metastasis-associated 1；Cervical cancer；Tumor migration 宫颈癌是最常见的女性恶性肿瘤之一，其发病率及病死率呈逐年上升的趋势[1]。侵袭、转移是影响宫颈癌病死率及预后的主要因素[2-4]。研究表明，MTA1在许多肿瘤中都表达上调。MTA1基因属于组蛋白去乙酰基酶（HDAC）家族的一个成分，能够通过改变组蛋白的乙酰化状态进行调控基因的表达，使得一些癌细胞向着更具有侵袭性的表型转化。然而，MTA1在宫颈癌中的研究却极少，因此我们选取了宫颈癌细胞HeLa检测其MTA1的表达情况，并且观察MTA1对其转移、侵袭的影响。 1材料与方法 1.1 材料 人宫颈癌细胞系HeLa购自美国典型物种保存中心（ATCC），置于含10%的胎牛血清、100U/mL链霉素及100U/mL青霉素的DMEM培养基（购自Gibco公司），在37℃、5%CO2培养箱中培养，24h更新新鲜培养基。0.25%胰蛋白酶消化，收集并传代。选取对数生殖期细胞用于实验。羊抗人MTA1多克隆抗体