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Studies on the Effects and Mechanism of Phe-DDPT on A549 Cells Apoptosis

Author: LiJiaWei
Tutor: ZhaoZhiGuang
School: Lanzhou University
Course: Botany
Keywords: Phe - ddpt Lung cancer Tubulin Apoptosis Bax Caspase3
CLC: R285.5
Type: Master's thesis
Year: 2010
Downloads: 52
Quote: 0
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Abstract


The observation of this thesis compares the novel podophyllotoxin derivative of N - ( 1 - oxo - 2 ,2,6,6 - tetramethyl -4 - piperidin- butyloxycarbonyl ) - L - phenylalanine 4'- demethyl - 4 - deoxy the podophyllotoxin ( Phe - ddpt ) and etoposide the glycosides ( VP - 16 ) induced apoptosis of human lung adenocarcinoma cell line A549 and its regulation mechanism discussed . The results the following conclusions : ( 1 ) the Phe - ddpt can inhibit the proliferation of A549 cells and concentration dependent , with the same group 36h comparison , the Phe - ddpt with the time the OD significantly reduced ( P lt ; 0.01 ) , 12h and 48h no significant changes in VP - 16 groups increased with time still significantly inhibited proliferation ( P lt ; 0.01 ) . At the same point in time , the Phe - ddpt compared with VP-16 , and each group was significantly inhibited cell proliferation, and the inhibitory ability of increasing concentrations , the OD value was significantly lower ( P lt ; 0.05 , P lt ; 0.01 ) ; (2) flow cytometry test Phe - ddpt treated cells, A549 cells in the G1 phase large block, indicating that the cell cycle arrest and apoptosis . (3) Immunofluorescence staining showed that presented by the the Phe - ddpt processing nuclear chromatin condensation, and fragmentation and apoptotic bodies . Through statistical analysis found that the apoptosis rate gradually increased with the drug concentration increases . ( 4 ) The results of agarose gel electrophoresis after the Phe-DDPT processing cells , cells of different sizes of the DNA fragment , Phe-DDPT effectively induce A549 cell apoptosis . Cells tubulin polymerization generates a large number of micro - tube ( 5 ) in A549 cells induced fluorescence microscopy electron microscope Phe-DDPT induced by the Phe - ddpt can microtubules generating inhibition of cell proliferation and apoptosis . ( 6 ) Western blot detected the Phe - ddpt can induce cell Bax , Caspase3 protein expression increased , decreased Bcl-2 protein expression . These results suggest that : good inhibitory effect of new podophyllotoxin derivatives the Phe-DDPT of A549 cells through regulation of Bax , Caspase3 , Bcl - 2 protein gene expression induced A549 cell cycle arrest in G1 . This inhibitory activity against tumor cells and low toxicity effects in the the pilot compound of antitumor screening has important significance.

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