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Trichostatin A in vitro effects of HeLa cells Spectroscopy

Author: YangZhanGuo
Tutor: LuoRongHui
School: Zhengzhou University
Course: Optics
Keywords: Trichostatin A HeLa cells Ultraviolet absorption spectrum Raman spectroscopy
CLC: R73-3
Type: Master's thesis
Year: 2009
Downloads: 17
Quote: 1
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Abstract

Spectroscopy is a highly sensitive means of detection, accurate analysis and understanding of the structure of matter, the components and their contents effective method suitable for the state of cells and biological molecules and molecular microenvironment of the physical and chemical characteristics research. Relative to other traditional research methods of biology, spectroscopy technique has the advantages of non-destructive, non-invasive, without reagents and a high degree of automation. HeLa cells are widely used in cancer research in laboratories around the world. The cell lines from normal cervical cells by the transformation of a human papilloma virus (HPV18: Human Papillomavirus18) from cancer cells and other cancer cells, HeLa cells abnormal rapid proliferation can be an unlimited number of split go on. Streptomycin trichostatin A (trichostatin A, TSA) metabolites nanomolar concentrations of specific inhibition of histone deacetylase, is an internationally recognized research acetylated reliable mode of cell. Trichostatin A for histone acetylation in tumor and normal tissue, transformed cells induced differentiation or apoptosis, so as to achieve the role of anti-tumor. Trichostatin A trichostatin A role in HeLa cells after UV absorption spectroscopy and Raman spectroscopy, designed from the molecular level to explore the mechanism of cancer cells for future clinical trichostatin A The application of the foundation to provide new ideas, and also for other potential anti-cancer drug screening. The papers carry out experimental research in three main areas, draw the following important conclusions: 1. Different doses 24h, 48h TSA treated HeLa cell morphology and proliferation changes. 200nmol / L TSA results show that the treated cells, compared with the form and amount of the control cells were significantly different. The study of different doses of TSA treatment 24h, 48h HeLa cells UV absorption spectra. The results showed that 200nmol / L of TSA HeLa cells can be maximum absorption value at 260nm. Studied different doses of TSA treatment 24h, 48h HeLa cells Raman spectroscopy. The results showed that the protein secondary structure and side-chain amino acids, bases, phosphate backbone of the lipids of the spectrum there is a certain relationship with the TSA dose: (1) TSA for 24h causing the vested intracellular protein secondary structure amide I of α-helical of 1658 cm ? ^{-1 and 938cm ^{-1 peak position of strength decline, but 200nmol / L dose of TSA is a relatively small impact. Characterization amide III rules curl 1241cm ^{-1 intensity of the peaks in the dosing samples show a downward trend, while 200nmol / L TSA handle relatively small influence. Tyrosine bending vibration in 200nmol / L TSA treated sample reaching the strongest, but also tends to decrease and then increase with TSA dose. An indole ring of tryptophan residues in 200nmol/LTSA treated samples Raman peak intensity of the highest peak is sharp, is \trend. (2) TSA treatment of cells 24h after cell 1094cm ^{-1 peak intensity was significantly reduced, while 200nmol/LTSA treated sample Raman peak intensity increase, when the dose is further increased, the peak position of the sample relative intensity reduced. Characterization of A and G 1342cm ^{-1 vibration 50nmol / L TSA treated samples no significant change 100nmol / L the TSA sample Raman peak intensity reaches the highest, with the further increase of the dose , Raman peak intensity will decrease. Vesting A and G, 1584 cm ^{-1 peak, TSA dose of 50nmol / L and 100nmol / L treated sample spectra, respectively redshift 3cm ^{-1 and 5cm ^{-1 , to 200nmol / L TSA treated samples back to 1585cm ^{-1 . Description low dose the TSA role of the DNA strand breaks, vibration weakened while larger doses TSA may make the interaction between the HeLa cell DNA and histones weakened DNA easier depolymerization. 1067cm ^{-1 (3) TSA-treated cells 24h after vesting lipids CC stretching vibration of the Raman characteristic frequency peak intensity decreased, particularly evident in the sample of large doses of TSA treatment. Lipids by the TSA, to reduce the degree of order, CC stretching vibration weakened. (4) cells after the TSA role 48h Raman spectral trend with 24h particular, relative to the control sample, the 100nmol/LTSA at 48h samples and 200nmol/LTSA of samples for 24h consistent spectral changes. Note the increase due to the the TSA role of time, the original higher doses to achieve the effect, at the lower doses of the treatment can occur. (5) 200nmol / L of TSA-treated cells 24h and 48h Raman spectra there are differences, that deal with the intensity of the peak in the 48-hour samples are generally lower than the samples for 24 hours, but 1314cm ^{-1 the peak intensity is enhanced. Because TSA role in the increase of time, the cell morphology becomes more irregular, so that the cells as a main component to the lipid membrane area increases, an increase in lipid content, coupled with the conformational change, leading to no rules Characterization CH bond in the lipid curl vibration of the Raman peaks intensity increases.}}}}}}}}}}}

Trichostatin A in vitro effects of HeLa cells Spectroscopy

Abstract

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