ZBED1促进肺腺癌增殖功能初步研究
发布时间：2019-01-04 12:40
【摘要】：[目的]肺癌全球高发,发病率及死亡率居高不下,但是其发病机制远未阐明。ZBED1是与细胞增殖相关的重要转录因子,然而其在肺癌中的作用尚不明确。本研究拟从细胞(A549)和人体组织(肺腺癌及癌旁正常肺组织)两方面,研究ZBED1与肺腺癌的相关性,证实ZBED1表达对肺腺癌细胞增殖生物学行为的影响,为进一步研究肺腺癌发病分子机理提供实验依据。[方法]利用苏木素-伊红染色法以及免疫组织化学染色技术检测肺腺癌及癌旁正常肺组织中ZBED1的表达;并统计分析ZBED1与临床病理参数的相关性;Westernblotting方法检测ZBED1在肺腺癌细胞株A549、PC-9、NCI-H1975、NCI-H2342、SPC-A-1、XLA-07 以及正常肺上皮细胞BEAS-2B中的表达水平;siRNA技术敲减肺腺癌细胞A549中ZBED1基因;MTS增殖实验以及流式细胞周期检测技术研究ZBED1与肺腺癌细胞增殖的关系;Westernblotting检测敲减ZBED1后,细胞周期相关蛋白的变化;流式细胞凋亡检测技术研究ZBED1与肺腺癌细胞凋亡的关系。[结果] ZBED1在肺腺癌组织中的表达显著高于癌旁正常肺组织; ZBED1在肺腺癌细胞系中普遍表达;在A549细胞系中表达最高;在肺腺癌细胞系A549中敲减ZBED1后,细胞增殖能力减弱;在肺腺癌细胞系A549中敲减ZBED1后,细胞阻滞在G0/G1期;- Westernblotting检测结果显示:细胞周期蛋白CyclinDl、CyclinD2、CyclinE2、CDK2、CDK6、P21 表达无明显变化,CyclinD3、CDK4、CyclinEl表达降低,P15、P27、P53表达升高;在肺腺癌细胞系A549中敲减ZBED1后,细胞凋亡无明显变化。[结论]ZBED1在肺腺癌中异常高表达;ZBED1参与细胞周期调控促进肺腺癌细胞增殖功能。
[Abstract]:[objective] Lung cancer has a high incidence, high morbidity and high mortality, but its pathogenesis is far from clear. ZBED1 is an important transcription factor associated with cell proliferation, but its role in lung cancer is not clear. In this study, we studied the relationship between ZBED1 and lung adenocarcinoma from two aspects: cell line (A549) and human tissue (lung adenocarcinoma and adjacent normal lung tissue), and confirmed the effect of ZBED1 expression on the biological behavior of proliferation of lung adenocarcinoma cells. To provide experimental basis for further study of the molecular mechanism of lung adenocarcinoma. [methods] the expression of ZBED1 in lung adenocarcinoma and adjacent normal lung tissues was detected by hematoxylin eosin staining and immunohistochemical staining, and the correlation between ZBED1 and clinicopathological parameters was analyzed statistically. Westernblotting method was used to detect the expression of ZBED1 in NCI-H1975NCI-H2342SPC-A-1 XLA-07 and normal lung epithelial cell line A549PC-9NCI-H1975and BEAS-2B in normal lung epithelial cells, siRNA technique knocked down the ZBED1 gene in lung adenocarcinoma cell line A549. MTS proliferation assay and flow cytometry were used to study the relationship between ZBED1 and lung adenocarcinoma cell proliferation. Westernblotting was used to detect the changes of cell cycle associated protein after knockout ZBED1. Flow cytometry was used to study the relationship between ZBED1 and lung adenocarcinoma cell apoptosis. [results] the expression of ZBED1 in lung adenocarcinoma tissue was significantly higher than that in adjacent normal lung tissue, the expression of ZBED1 in lung adenocarcinoma cell line was universal, the highest expression was in A549 cell line, and the proliferation ability of lung adenocarcinoma cell line A549 was decreased after knockout of ZBED1. After knockout of ZBED1 in lung adenocarcinoma cell line A549, the cell was blocked in G0/G1 phase. The results of Westernblotting showed that the expression of cyclin CyclinDl,CyclinD2,CyclinE2,CDK2,CDK6,P21 did not change significantly, but the expression of CyclinD3,CDK4,CyclinEl decreased, and the expression of P15P _ (27) P _ (27) P _ (53) increased. The apoptosis of lung adenocarcinoma cell line A549 did not change after knockout of ZBED1. [conclusion] ZBED1 is overexpressed in lung adenocarcinoma and ZBED1 is involved in the regulation of cell cycle to promote the proliferation of lung adenocarcinoma cells.
【学位授予单位】：昆明医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R734.2
[Abstract]:[objective] Lung cancer has a high incidence, high morbidity and high mortality, but its pathogenesis is far from clear. ZBED1 is an important transcription factor associated with cell proliferation, but its role in lung cancer is not clear. In this study, we studied the relationship between ZBED1 and lung adenocarcinoma from two aspects: cell line (A549) and human tissue (lung adenocarcinoma and adjacent normal lung tissue), and confirmed the effect of ZBED1 expression on the biological behavior of proliferation of lung adenocarcinoma cells. To provide experimental basis for further study of the molecular mechanism of lung adenocarcinoma. [methods] the expression of ZBED1 in lung adenocarcinoma and adjacent normal lung tissues was detected by hematoxylin eosin staining and immunohistochemical staining, and the correlation between ZBED1 and clinicopathological parameters was analyzed statistically. Westernblotting method was used to detect the expression of ZBED1 in NCI-H1975NCI-H2342SPC-A-1 XLA-07 and normal lung epithelial cell line A549PC-9NCI-H1975and BEAS-2B in normal lung epithelial cells, siRNA technique knocked down the ZBED1 gene in lung adenocarcinoma cell line A549. MTS proliferation assay and flow cytometry were used to study the relationship between ZBED1 and lung adenocarcinoma cell proliferation. Westernblotting was used to detect the changes of cell cycle associated protein after knockout ZBED1. Flow cytometry was used to study the relationship between ZBED1 and lung adenocarcinoma cell apoptosis. [results] the expression of ZBED1 in lung adenocarcinoma tissue was significantly higher than that in adjacent normal lung tissue, the expression of ZBED1 in lung adenocarcinoma cell line was universal, the highest expression was in A549 cell line, and the proliferation ability of lung adenocarcinoma cell line A549 was decreased after knockout of ZBED1. After knockout of ZBED1 in lung adenocarcinoma cell line A549, the cell was blocked in G0/G1 phase. The results of Westernblotting showed that the expression of cyclin CyclinDl,CyclinD2,CyclinE2,CDK2,CDK6,P21 did not change significantly, but the expression of CyclinD3,CDK4,CyclinEl decreased, and the expression of P15P _ (27) P _ (27) P _ (53) increased. The apoptosis of lung adenocarcinoma cell line A549 did not change after knockout of ZBED1. [conclusion] ZBED1 is overexpressed in lung adenocarcinoma and ZBED1 is involved in the regulation of cell cycle to promote the proliferation of lung adenocarcinoma cells.
【学位授予单位】：昆明医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R734.2
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