ZBED1促進(jìn)肺腺癌增殖功能初步研究
發(fā)布時(shí)間：2019-01-04 12:40
【摘要】：[目的]肺癌全球高發(fā),發(fā)病率及死亡率居高不下,但是其發(fā)病機(jī)制遠(yuǎn)未闡明。ZBED1是與細(xì)胞增殖相關(guān)的重要轉(zhuǎn)錄因子,然而其在肺癌中的作用尚不明確。本研究擬從細(xì)胞(A549)和人體組織(肺腺癌及癌旁正常肺組織)兩方面,研究ZBED1與肺腺癌的相關(guān)性,證實(shí)ZBED1表達(dá)對(duì)肺腺癌細(xì)胞增殖生物學(xué)行為的影響,為進(jìn)一步研究肺腺癌發(fā)病分子機(jī)理提供實(shí)驗(yàn)依據(jù)。[方法]利用蘇木素-伊紅染色法以及免疫組織化學(xué)染色技術(shù)檢測(cè)肺腺癌及癌旁正常肺組織中ZBED1的表達(dá);并統(tǒng)計(jì)分析ZBED1與臨床病理參數(shù)的相關(guān)性;Westernblotting方法檢測(cè)ZBED1在肺腺癌細(xì)胞株A549、PC-9、NCI-H1975、NCI-H2342、SPC-A-1、XLA-07 以及正常肺上皮細(xì)胞BEAS-2B中的表達(dá)水平;siRNA技術(shù)敲減肺腺癌細(xì)胞A549中ZBED1基因;MTS增殖實(shí)驗(yàn)以及流式細(xì)胞周期檢測(cè)技術(shù)研究ZBED1與肺腺癌細(xì)胞增殖的關(guān)系;Westernblotting檢測(cè)敲減ZBED1后,細(xì)胞周期相關(guān)蛋白的變化;流式細(xì)胞凋亡檢測(cè)技術(shù)研究ZBED1與肺腺癌細(xì)胞凋亡的關(guān)系。[結(jié)果] ZBED1在肺腺癌組織中的表達(dá)顯著高于癌旁正常肺組織; ZBED1在肺腺癌細(xì)胞系中普遍表達(dá);在A549細(xì)胞系中表達(dá)最高;在肺腺癌細(xì)胞系A(chǔ)549中敲減ZBED1后,細(xì)胞增殖能力減弱;在肺腺癌細(xì)胞系A(chǔ)549中敲減ZBED1后,細(xì)胞阻滯在G0/G1期;- Westernblotting檢測(cè)結(jié)果顯示:細(xì)胞周期蛋白CyclinDl、CyclinD2、CyclinE2、CDK2、CDK6、P21 表達(dá)無(wú)明顯變化,CyclinD3、CDK4、CyclinEl表達(dá)降低,P15、P27、P53表達(dá)升高;在肺腺癌細(xì)胞系A(chǔ)549中敲減ZBED1后,細(xì)胞凋亡無(wú)明顯變化。[結(jié)論]ZBED1在肺腺癌中異常高表達(dá);ZBED1參與細(xì)胞周期調(diào)控促進(jìn)肺腺癌細(xì)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.
【學(xué)位授予單位】：昆明醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：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.
【學(xué)位授予單位】：昆明醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：R734.2
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