Imaging of P-glycoprotein of H69/VP small-cell lung cancer lines by scanning near-field optical microscopy and confocal laser microspectrofluorometer.
Chemoresistance remains the major obstacle to successful therapy of the lung cancer. The multi-drug resistance (MDR) is generally associated with altered expression of drug transporter proteins, such as P-glycoprotein (P-gp). So the distribution of P-gp on the membrane is of great importance to further study the interaction between drug and P-gp. In the present work, the P-gp of the H69/VP small-lung cancer cells was detected using monoclonal antibody UIC2. A secondary goat-anti mouse antibody coupled with biotin was used. The fluorescence emission was detected from a streptavidin-Texas Red. Results were investigated by a homemade scanning near-field optical microscope (SNOM) coupled to a confocal laser microspectrofluorometer (CLMF). Topographical images and localized spectra were obtained at the level of one cell membrane. It was found that the distribution of P-gp is not homogeneous and this observation is basically in accord with the fluorescent images obtained by classical microscopy. The distribution of P-gp would be localized in a higher region on a cell surface. This methodology would also enhance our understanding of MDR under physiological conditions.
Qiao
Weihong
W
Unité MéDIAN, CNRS-UMR 6142, UFR de Pharmacie, IFR 53, Université de Reims Champagne-Ardenne, 51 rue Cognacq Jay, 51096 REIMS Cédex, France. qiaoweihong@hotmail.com
Shang
Guangyi
G
Lei
Franck H
FH
Trussardi-Regnier
Aurélie
A
Angiboust
Jean-F
JF
Millot
Jean-M
JM
Manfait
Michel
M
D018435:ATP Binding Cassette Transporter, Subfamily B / Q000378:metabolism*; D018288:Carcinoma, Small Cell / Q000378:metabolism / Q000648:ultrastructure*; D045744:Cell Line, Tumor; D006801:Humans; D008175:Lung Neoplasms / Q000378:metabolism / Q000648:ultrastructure*; D018613:Microscopy, Confocal / Q000295:instrumentation / Q000379:methods; D008855:Microscopy, Electron, Scanning / Q000295:instrumentation* / Q000379:methods*
D018435:ATP Binding Cassette Transporter, Subfamily B