Regulating The Function Of CD40 Expression And Significance. CD40 And Its Ligand CD154 (CD40L) Connection For Lung Cancer

Objective: To determine the possible functional significance of CD40-CD40 ligand (CD154) engagemet on human lung carcinomas and to assess the potetial of CD40 as a therapentic target.Method: ① CD40-negative cell lines (GLC-82) were screeed by flow cytometry. I constructed prokaryocyte expression vector pCDNA3-CD40 CDNA which was used to transfect the GLC-82 cell lines, I got CD40-transfected cell lines GLC-82/CD40. ZK618 and FCH712 cell lines were established by culturing tissue imprints derived from fresh tumor biopsy tissues from 2 patients of lung cancer. 0.1 μ g/ml of CD40L (CD154) was added to medium of 7 lung tumor cell lines, including GLC-82, A549, NCI-H209, CALU-3, ZK618, FCH712 and GLC-82/CD40. The cells were cultured for l-5d. The cells wre harvested and analysed by flow cytometry. We evaluated the effect of CD40L on the surface expression of MHC-I, Fas, bcl2, ICAM-1,EGFR, PGP and LRP, and the cell cycle was analysed with propidium iodiode (PI) assay, then the cell apoptosis with anexinV assay. MTT assay was used to evaluation of the effect of CD40L on the growth kinetics of the lung cancer cell lines. ② We obtained 38 surgical biopsy specimens from lung cancers, 25 nornal lung specimens next to turmor position, and 5 non-malignant tussue specimens. The surface expression of CD40, MHC-I, EGFR, ICAM-1, PGP, LRP, P53, bcl2 and cell cycle on these tissues were determined by flow cytometric cell phenotyping analysis and PI staining assay.Results:① GLC-82 cell line was negative for CD40. NCI-H209 exhibited a low percentage of CD40. A549 was scored as moderatedly positive for CD40. CALU-3, ZK618 and FCH712 cell lines expressed high levels of CD40, the CD40cDNA transfected cell line GLC-82/CD40 also expressed high levels of CD40 (95.9%). All cell lines exhibited high percentage of MHC-I except NCI-H209. All cell lines also expressed EGFR, and others...