RASSF1A基因对乳腺癌细胞恶性生物学特征的作用研究  

Effect of RASSF1A gene on the malignant biological characteristics of breast cancer cells

作　　者：郑璐[1] 汤铜[1] 杜旺 贾文俊 张磊[1] 钱波[1] 李佳[1] 黄赞 丁一博 Zheng Lu;Tang Tong;Du Wang(Dept of General Surgery,The Second Affiliated Hospital of Anhui Medical University,Hefei 230601)

机构地区：[1]安徽医科大学第二附属医院普外科

出　　处：《安徽医科大学学报》2019年第6期882-886,共5页Acta Universitatis Medicinalis Anhui

基　　金：安徽省自然科学基金(编号:1308085QH152);安徽医科大学校科研基金联合资助项目(编号:2015xkj111、2018xkj038);安徽省高等学校省级质量工程项目(编号:2014jyxm110)

摘　　要：目的通过脂质体介导细胞转染,构建Ras相关区域家族1(RASSF1A)基因过表达的乳腺癌MCF-7细胞株,研究RASSF1A基因对乳腺癌MCF-7细胞株增殖、克隆形成、凋亡的作用。方法构建RASSF1A过表达载体并鉴定,通过脂质体介导细胞转染将空载体及含有目的基因的重组载体转染到乳腺癌MCF-7细胞株中。分别通过荧光显微镜观察、实时荧光定量PCR(RT-PCR)、Western Blot观察转染效率及确定最优转染条件。并通过四甲基偶氮唑盐法(MTT)、平板克隆、流式细胞术分别检测RASSF1A重组基因转染组、空载体转染组及空白细胞对照组细胞增殖、细胞克隆形成以及细胞凋亡的情况。结果转染RASSF1A基因后的乳腺癌MCF-7细胞株出现该基因mRNA及蛋白质水平显著提升,证明RASSF1A基因过表达的MCF-7细胞株构建成功。MTT、平板克隆及流式细胞术检测结果显示:与对照组相比,RASSF1A基因过表达的MCF-7细胞株增殖、克隆形成能力受到抑制,而细胞凋亡率增加。结论成功构建了RASSF1A基因过表达的MCF-7细胞株,RASSF1A基因可能参与了乳腺癌MCF-7细胞株增殖、凋亡的调控,其机制可能与抑制BC细胞增殖、促进凋亡有关。Objective To construct a breast cancer MCF-7 cell line overexpressing Ras-related region family 1(RASSF1A) gene,and study the effect of RASSF1A gene on proliferation,colony formation and apoptosis of breast cancer MCF-7 cell line. Methods RASSF1A over expression vector was constructed and identified.The empty vector and recombinant vector containing the target gene was transfected into breast cancer MCF-7 cell line by liposome mediated transfection.The transfection efficiency was observed by fluorescence microscopy,real-time quantitative PCR and Western blot,and the optimal transfection conditions were determined.MTT,Plate clone formation assay and Flow cytometry were used to detect the cell proliferation,cell clone formation and apoptosis of RASSF1A recombinant gene transfection group,empty vector transfection group and blank control group. Results After transfection of RASSF1A gene,the mRNA and protein levels of breast cancer MCF-7 cell line were significantly increased,which proved that the RASSF1A gene over expression of MCF-7 cell line was successfully constructed.MTT,Plate clone formation assay and Flow cytometry showed that compared with the control group,the proliferation and cloning ability of MCF-7 cell line overexpressing RASSF1A gene was inhibited,and the apoptosis rate was increased. Conclusion The MCF-7 cell line overexpressing RASSF1A gene is successfully constructed.RASSF1A gene may be involved in the regulation of proliferation and apoptosis of breast cancer cell line MCF-7.The mechanism may be related to the inhibition of proliferation and apoptosis of breast cancer cells.

关 键 词：RASSF1A基因 乳腺癌 MCF-7 增殖凋亡 

分 类 号：R346[医药卫生—基础医学]