摘要
目的探讨Ⅰ型胶原α1链(COL1A1)基因沉默对人乳腺癌MDA-MB-231细胞侵袭与转移的影响。方法设计2条shRNA干扰载体及1条阴性对照载体,分别稳定转染乳腺癌MDA-MB-231细胞。用Western blot法筛选抑制效率最高的细胞进行细胞平板集落形成实验、细胞黏附实验、细胞迁移及侵袭实验,观察COL1A1基因对MDA-MB-231细胞黏附、运动及侵袭能力的影响。结果转染pshRNA-COL1A1-2干扰载体的MDA-MB-231细胞COL1A1蛋白表达降低最明显,抑制率达66.98%±2.08%,选择该组细胞作为有效干扰组进行后续实验。pshRNA-COL1A1转染组细胞的集落形成率显著低于空白对照组和阴性质粒pshRNA-scramble转染组(P<0.05);细胞黏附实验中pshRNA-COL1A1转染组细胞的吸光度值较对照组明显降低(P<0.001);转染pshRNA-COL1A1质粒组细胞的穿膜细胞数也显著低于空白对照组和阴性质粒pshRNA-scramble转染组(P<0.001)。结论 COL1A1基因沉默可在体外抑制人乳腺癌MDA-MB-231细胞的黏附、侵袭和迁移。
Objective To investigate the effects of shRNA-mediated human collagen type 1 alpha 1 （ COL1A1 ） gene silence on the migration and invasion of Breast Cancer Cell Line MDA-MB-231. Methods Two shRNA vectors and one negative control vector were designed and stably transfected into MDA-MB-231 ceils. Western blot analysis was used to screen for the group which had the highest inhibitory rate. Monolayer colony formation assay was performed to assess a single cell proliferation ability. The cell adhesion, migration and invasion potencies were observed by cell-matrix adhesion assay, and Transwell assay. Results Cells transfected with pshRNA-COL1A1-2 vector had the stronger inhibition of COL1A1 protein, with the inhibition rate being 66.98%±2. 08%, and cells in this group were used for the subsequent experiments. Compared with control group and pshRN er colony formation rate decreased obviously （P 〈 0. 05） . The optical absorbance A-scramble group, the monolay- value of adherent cells and the number of invading and migrating ceils which moved across the matrix barrier for pshRNA-COL1 A1 group were less than that of control group and pshRNA-scramble group （ P 〈 O. 001 ）. Conclusions COL1A1 gene silencing of hu- man MDA-MB-231 cells may inhibit cell matrix adhesion, migration and invasion potencies in vitro.
出处
《基础医学与临床》
CSCD
北大核心
2013年第8期981-985,共5页
Basic and Clinical Medicine
基金
广东省医学科研基金(B2012203)
广东省领军人才专项基金(C1030925)
