Prostate cancer-specific molecular probes of the building and its experimental study in vivo MR imaging

Title: Prostate cancer-specific molecular probes of the building and its experimental study in vivo MR imagingAuthor: Ren JingDegree-granting units: Fourth Military Medical UniversityKeywords: molecular imaging;; prostate cancer;; prostate stem cell antigen;; monoclonal antibody;; magnetic nano-gold particles;; magnetic resonance imagingAbstract:

 Purpose:

 1 of cultured human prostate cancer cell lines, Neodymium Magnets the establishment of prostate cancer nude mice model, in which the expression of PSCA and
Its meaning;

 (2) application of nano-gold labeled anti-human PSCA magnetic particles monoclonal antibody 7F5, build a prostate cancer-specific molecular probes MR
7F5 @ GoldMag, detection of prostate cancer cells with binding specificity;

 3 of 7F5 @ GoldMag for in vitro, in vivo MR imaging
The feasibility and specificity in PCa diagnosis and significance.

 Method:

 1 of cultured human prostate cancer cell line LNCaP,
PC-3 and human hepatoma cell line SMMC-7721, the establishment of nude models, using immunohistochemistry, indirect immunofluorescence, cells of the immune
Science, such as flow cytometry and western blotting method to detect PSCA in vitro prostate cancer cell lines and tumors in nude
Expression.

 (2) the gold particles as MR contrast http://www.everbeenmagnet.com/en/products/110-sintered-neodymium-magnets agent magnetic markers 7F5, application of non-covalent coupling methods to build prostate cancer-specific
MR molecular probe 7F5 @ GoldMag, by laser confocal microscopy, flow cytometry, transmission electron microscopy and other methods, detection probe
Combined with the specificity of prostate cancer.

 3. The use of clinical applications 3.0T magnetic resonance scanner to establish MR scanning sequence, the observations of
Concentration of gold particles with magnetic MR imaging, to explore the MR can distinguish the lowest concentration; of probes for specific imaging of viable cells in vitro
Sex; of tumor-bearing nude mice by intravenous injection of 7F5 @ GoldMag, respectively, before injection, after injection, 6 h, 12 h and 24 h-line MR scan, observe
Its effects on tumor T2WI signal, used to explore the feasibility of in vivo MR imaging of tumors.

 Results:

 1 human prostate cancer cells
Cell lines LNCaP, PC-3 and the human hepatoma cell line SMMC-7721 were single-adherent growth; LNCaP cells into the tumor rate of 12.5% ​​(1 / 8); PC-3 and
SMMC-7721 cells into the tumor rate of 100% (8 / 8); immune staining, indirect immunofluorescence, Western-blot and flow cytometry
Test results confirmed that LNCaP, PC-3 cells express PSCA; SMMC-7721 cells do not express PSCA; nude tissue immunohistochemistry
Staining confirmed similar results.

 (2) to build a successful MR prostate cancer-specific molecular probe 7F5 @ GoldMag; flow cytometry
The results 7F5 @ GoldMag and LNCaP and PC-3 cells were combined with 85.2% and 92.1%, while combined with the rate of SMMC-7721 cells
4.2%, independent of antibody @ GoldMag group three types of cells were negative; laser confocal microscope see LNCaP and PC-3 cells with
7F5 @ GoldMag a specific combination of red fluorescence in the cell membrane, SMMC-7721 cells, no red fluorescent membrane, independent of antibody group
The cell lines were negative; TEM LNCaP and PC-3 near the cell membrane can be seen more than a diameter of about 50nm dense electron density of the agglomerate
Degree of particle aggregation, SMMC-7721 cell surface was no compact electron density particles, independent of antibody @ GoldMag group of individual cell lines are only as
Nomi a small amount of electron density in dense granules, and the farther away from the cell membrane.

 3. GoldMagTM-CS was diluted to 1:640 with 1% agarose gel
Compared to plastic, T2WI sequence signal intensity difference was statistically significant; using 7F5 @ GoldMag line labeled MR scanning of different cell lines results suggest that
7F5 @ GoldMag can significantly reduce LNCaP and PC-3 cells T2WI signal, while the SMMC-7721MR signal no significant effect. Non-specific antibody
Coupled GoldMag MR signal of the three types of cells had no significant effect.

 4 MR imaging results of nude mice, PC-3 tumor-bearing nude mice injected
7F5 @ GoldMag 6 h, 12 h and 24 h after the T2WI signal intensity of tumor tissue was significantly lower than the plain
(ANOVA, F = 43.675, p = 0.000); 12 h after injection of the probe signal intensity over 6 h to further reduce (p = 0.001); and 12 h after injection
24 h the tumor signal intensity difference was not statistically significant (p = 0.145). The SMMC-7721 +7 F5 @ GoldMag, PC-3 + irrelevant antibody
@ GoldMag and SMMC-7721 + @ GoldMag group unrelated antibody scan and injection of contrast medium 6 h, 12 h and 24 h the signal intensity of tumor
The difference was not statistically significant.

 Conclusion:

 Application of gold nanoparticles with magnetic particles conjugated anti-PSCA monoclonal antibody 7F5 was successfully constructed prostate
Cancer-specific MR probe 7F5 @ GoldMag, with good physicochemical properties and immunological activity in the 3.0 T MR scanner detects the forefront of in vitro
Adenocarcinoma cells and in vivo targeting of prostate cancer with enhanced effects, clinical trials for the next step and lay the foundation.Degree Year: 2009