仇静 张月 陈书兰 刘岚 刘峤峤 张广耘 袁晓△.腺病毒携带shFOXM1 对ACC-2 细胞作用机制的研究[J].,2016,16(7):1249-1252 |
腺病毒携带shFOXM1 对ACC-2 细胞作用机制的研究 |
Research on the Mechanismof ACC-2 Cell Regulation by AdenovirusHarboring shFOXM1 |
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DOI: |
中文关键词: 叉头框蛋白M1 人涎腺腺样囊性癌 细胞周期 |
英文关键词: Forkhead box protein M1 Adenoid cystic carcinoma Cell cycle |
基金项目:国家自然科学基金项目(31170891) |
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中文摘要: |
目的:研究携带特异性抑制叉头框蛋白M1(Forkhead box protein M1,FOXM1)表达的腺病毒(Ad5.shFOXM1)对人涎腺腺样
囊性癌(Adenoidcystic carcinoma,ACC)细胞周期的调控、细胞凋亡的影响及其作用机理。方法:根据人FOXM1 mRNA的序列,设
计合成针对FOXM1 基因的三对shRNA,转染ACC-2 细胞系,利用实时荧光定量PCR、Western blot 筛选获得最佳干扰片段,并构
建入腺病毒载体,利用流式细胞术、MTT、Western blot等检测其对ACC-2 细胞周期、细胞凋亡的调控及分子作用。结果:①成功筛
选获得了抑制FOXM1 表达的shRNA,并构建了特异性抑制ACC-2 细胞中FOXM1 表达的腺病毒Ad5-shFOXM1;②
Ad5-shFOXM1 能明显抑制ACC 细胞的增殖,并引起ACC-2 细胞S期的阻滞(P<0.05);②Ad5-shFOXM1 通过下调c-Myc 蛋白的
表达抑制了细胞的增殖,通过下调P21Cip1,P27Kip1 蛋白的表达抑制了ACC-2 细胞周期(P<0.05)。结论:下调FOXM1的表达能
够明显抑制ACC-2 细胞周期和ACC-2 细胞的增殖。 |
英文摘要: |
Objective:Research on the specific inhibition of forkhead box protein M1 (FOXM1) expression by adenovirus (Ad5.
shFOXM1) and the mechanismof cell cycle and cell apoptosis regulation in human adenoidcystic carcinoma (ACC).Methods:According
to the sequence of human FOXM1 mRNA, we designed and synthesis three FOXM1 shRNA, which was transfected into ACC-2 cells.
We identified the most potently suppressed FOXM1 mRNA by Real-time PCR and Western blot. We constructed adenovirus vector harboring
the best interference fragment, and detected the cell cycle and cell apoptosis regulation and mechanism by flow cytometry, MTT,
Western blot in ACC-2.Results:We identified the most potent shRNA targeting FOXM1. After constructing of Ad5.shFOXM1, the Ad5.
shFOXM1 can significantly inhibit the proliferation of ACC-2 cells, and caused the block of ACC-2 cells in the S phase (P<0.05);
Ad5-shFOXM1 inhibited cell proliferation through down regulating the expression of c-Myc, and down regulating the expression of
P21Cip1, P27Kip1 protein expression in ACC-2 cell (P<0.05).Conclusion:Down-regulation of FOXM1 expression can inhibit the cell
cycle and cell proliferation of ACC. |
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