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[MADD在肺腺癌组织中的表达及其对肺腺癌A549细胞增殖和凋亡的影响]

[Expression of MADD in lung adenocarcinoma tissues and its effects on proliferation and apoptosis of lung adenocarcinoma A549 cells].

作者信息

Wei Yu-Ping, Wu Jin-Xiang, Guo Yuan-Fang, Sun Gao-Ying, Zhang Qiang, Bi Wen-Xiang, Dong Liang

机构信息

Institute of Biochemistry and Molecular Biology, School of Medicine, Qilu Hospital, Shandong University, Jinan 250012, China.

出版信息

Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi. 2012 Apr;28(4):354-7.

Abstract

AIM

To investigate the expression of MAPK-activating death domain protein (MADD) in lung adenocarcinoma tissues and its effects on proliferation and apoptosis of lung adenocarcinoma A549 cells.

METHODS

Immunohistochemistry was used to detect the expression of MADD in lung normal and tumor tissues. The expression of IG20 gene in A549 cells was measured by reverse transcription polymerase chain reaction. A549 cells were transfected with pEYFP-MADD plasmids carrying MADD gene or pNL-SIN-GFP-MID lentiviral vectors used for RNA interference. MADD expression and cell proliferation and apoptosis were determined by Western blot, MTT assay, and flow cytometry.

RESULTS

The expression levels of MADD were higher in lung adenocarcinoma and squamous cell carcinoma tissues than that in lung normal tissues, and lung adenocarcinoma tissues expressed more MADD than lung squamous cell carcinoma tissues. The transcript encoding MADD was expressed in A549 cells. The transfection of pEYFP-MADD plasmids could increase MADD expression and cell proliferation of A549 cells, while the A549 cells transfected with pNL-SIN-GFP-MID lentiviral vectors showed significantly decreases in the MADD level and proliferation. It is shown that MADD overexpression could inhibit A549 cell apoptosis, and knock down of MADD could promote apoptosis of them.

CONCLUSION

The expression of MADD increases obviously in lung adenocarcinoma, and MADD can promote survival of lung adenocarcinoma cells by inhibiting apoptosis.

摘要

目的

探讨丝裂原活化蛋白激酶激活死亡结构域蛋白(MADD)在肺腺癌组织中的表达及其对肺腺癌A549细胞增殖和凋亡的影响。

方法

采用免疫组织化学法检测MADD在肺正常组织和肿瘤组织中的表达。通过逆转录聚合酶链反应检测A549细胞中IG20基因的表达。用携带MADD基因的pEYFP - MADD质粒或用于RNA干扰的pNL - SIN - GFP - MID慢病毒载体转染A549细胞。通过蛋白质免疫印迹法、MTT法和流式细胞术检测MADD表达及细胞增殖和凋亡情况。

结果

MADD在肺腺癌和肺鳞癌组织中的表达水平高于肺正常组织,且肺腺癌组织中MADD的表达高于肺鳞癌组织。编码MADD的转录本在A549细胞中表达。转染pEYFP - MADD质粒可增加A549细胞中MADD的表达和细胞增殖,而用pNL - SIN - GFP - MID慢病毒载体转染的A549细胞中MADD水平和增殖明显降低。结果表明,MADD过表达可抑制A549细胞凋亡,敲低MADD可促进其凋亡。

结论

MADD在肺腺癌中的表达明显增加,且MADD可通过抑制凋亡促进肺腺癌细胞存活。

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