| Objective:To observe the surface structure of epithelial cells from normal breast and breast hyperplasia and breast cancer by atomic force microscope,to study the mechanism of breast carcinoma,to obtain the parameters and find out the regularity of membrane protein distribution of mammary epithelial cells,and to guide the clinical diagnosis of early breast cancer.Methods:Specimens of mammary hyperplasia,breast cancer tissue and normal breast tissue were collected from general hospital of Chinese People's Armed Police Force,15 cases in each group.Each specimen was fixed with 2%formalin and made into print slices,then was dried in clean space.The prepared samples were put on platform of AFM,were observed under Nikon inverted microscope for the dispersion of cells.10 cells of each specimen were chosen and 10 parts of each cell were scanned.Tapping mode was selected,using tapping mode in the range of 30μm～1μm and at the rate of 0.5Hz～2Hz.AFM image analysis software was used to obtain the parameters of membrane surface structure in order to find out the difference in the above-mentioned specimens.Average roughness(Ra),Peak count and Max height-depth(Rp-v)were detected by the AFM image software.Kruskal Wallis Test was carried out for statistical evaluation.Significance was set at P< 0.05.Results:The cellular membrane surface morphous and structure were observed clearly by the AFM:The membrane surface of normal breast was smooth,with a few of apophysis on the membrane.The cellular membrane surface of mammary gland hyperplasia was rough,with more apophysis on the membrane.The cellular membrane surface of breast cancer has distribution-intensive apophysis and some of the apophysis perform the phenomenon of convergence.The parameters of the cellular membrane surface were detected by AFM image analysis software,150 cells in each group.The Ra,peak count and Rp-v were 9.554±2.001nm, 13.180±4.040 nm,33.895±7.347 nm in the group of normal breast.The Ra,peak count and Rp-v were 14.730±3.945 nm, 15.647±5.583nm,47.527±14.014 nm in the group of breast hyperplasia.The Ra,peak count and Rp-v were 23.040±6.727 nm, 16.560±6.227nm,78.436±35.841nm in the group of breast cancer.From the statistical Analysis of the parameters mentioned above,Ra and Rp-v were significantly different among the three groups(P <0.01)Peak count was not significantly different between hyperplasia and breast cancer(P>0.05).The measurement and analysis of cellular membrane protein distribution with AFM is of great significance for the single-cell diagnosis of breast cancer.The statistic analysis of Ra,peak count and Rp-v shows that the cellular membrane protein increase gradually in the progression from normal mammary epithelial cells to hyperplasia and breast cancer.This is relative to its function,for the protein the cellular membrane is getting more and more active from normal breast to breast cancer cells.Conclusion:In this experiment,the membrane surface structure of mammary epithelial cells was observed with AFM first in the world.It is found that In the progress from normal lacteal gland to hyperplasia gland,the fine structure of cellular membrane changed accordingly,and the membrane proteins increased gradually,with apophysis increasing gradually shown by AFM.When the normal mammary gland becomes hyperplasia gland,the proteins increase in the cellular membrane.When breast cancer occurs,the proteins in the cellular membrane change variously,showing that the apophysis on the cellular membrane increase d and the diameters of them increased,with some showing fusion trend.The experiments analyzed the surface structure of breast epithelial cellular by AFM first in the world,proved that the diagnosis of breast cancer could be judged earlier by the observation of the changes in the cellular membrane,and single cell diagnosis could be made with AFM.
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