Xueying Mao

Isolation of circulating tumor cells (CTCs) from peripheral blood has the potential to provide a far easier "liquid biopsy" than tumor tissue biopsies, to monitor tumor cell populations during disease progression and in response to therapies. Many CTC isolation technologies have been developed. We optimized the Parsortix system, an epitope independent, size and compressibility-based platform for CTCs isolation, making it possible to harvest CTCs at the speed and sample volume comparable to standard CellSearch system. We captured more than half of cancer cells from different cancer cell lines spiked in blood samples from healthy donors using this system. Cell loss during immunostaining of cells transferred and fixed on the slides is a major problem for analyzing rare cell samples. We developed a novel cell transfer and fixation method to retain >90% of cells on the slide after the immunofluorescence process without affecting signal strength and specificity. Using this optimized method, we evaluated the Parsortix system for CTC harvest in prostate cancer patients in comparison to immunobead based CTC isolation systems IsoFlux and CellSearch. We harvested a similar number (p = 0.33) of cytokeratin (CK) positive CTCs using Parsortix and IsoFlux from 7.5 mL blood samples of 10 prostate cancer patients (an average of 33.8 and 37.6 respectively). The purity of the CTCs harvested by Parsortix at 3.1% was significantly higher than IsoFlux at 1.0% (p = 0.02). Parsortix harvested significantly more CK positive CTCs than CellSearch (p = 0.04) in seven prostate cancer patient samples, where both systems were utilized (an average of 32.1 and 10.1 respectively). We also captured CTC clusters using Parsortix. Using four-color immunofluorescence we found that 85.8% of PC3 cells expressed EpCAM, 91.7% expressed CK and 2.5% cells lacked both epithelial markers. Interestingly, 95.6% of PC3 cells expressed Vimentin, including those cells that lacked both epithelial marker expression, indicating epithelial-to-mesenchymal transition. CK-positive/Vimentin-positive/CD45-negative, and CK-negative/Vimentin-positive/CD45-negative cells were also observed in four of five prostate cancer patients but rarely in three healthy controls, indicating that Parsortix harvests CTCs with both epithelial and mesenchymal features. We also demonstrated using PC3 and DU145 spiking experiment that Parsortix harvested cells were viable for cell culture.

Prostate cancer is significantly more common in Western men than in Asian men, but the basis for this difference remains unknown. Because genomic studies of Asian prostate cancer are very limited, we used a genome-wide approach to reveal the genomic alterations in Chinese prostate cancers. We found a significant reduction in the frequency of certain somatic genomic changes that are commonly found in Western prostate cancers, including the 21q22.2-22.3 deletion, which involves the TMPRSS2:ERG fusion gene, and 10q deletion, which causes PTEN inactivation. Array results were confirmed by PCR-based molecular copy-number counting in selected samples. The different frequencies of these genomic changes were further evaluated by fluorescent in situ hybridization and immunohistochemistry analyses of tissue microarray samples. These alterations might be key genetic changes underlying the regional/ethnic difference in clinical incidence and might be induced by specific environmental and/or genetic risk factors that Western men are exposed to. Our findings suggest that tumors arise in Western and Chinese populations by alternative pathogenetic mechanisms.

ABSTRACT This study was conducted to investigate the free-radical-scavenging activity and antioxidant activity of silk sericin. Silk sericin was prepared from silkworm Bombyx mori and its ability to scavenge hydroxyl, superoxide and 1,1-diphenyl-2-picrylhydrazyl (DPPH) radicals was determined by Electron Spin Resonance (ESR) and ultraviolet spectrophotometry, respectively. The antioxidant activities of the silk sericin, including lipid peroxidation in the linoleic acid system, reducing power and ferrous-ion-chelating ability, were evaluated. The results showed that silk sericin had a strong scavenging capacity for hydroxyl, superoxide and DPPH radicals. The results also showed that silk sericin had potent antioxidative activity on the peroxidation of linoleic acid. The reducing power and ferrous-ion-chelating ability of silk sericin were significant. These results indicated that silk sericin from silkworm B. mori was a natural antioxidant with potent antioxidative activity. PRACTICAL APPLICATIONS Most of silk sericin must be removed during raw silk production at the reeling mill and other stages of silk processing. At present, silk sericin is mostly discarded in silk processing waste water. If silk sericin is recovered and recycled, it can represent a significant economic and social benefit. Silk sericin is a natural macromolecular protein derived from the silkworm Bombyx mori. Silk sericin is useful because of its antioxidant activity. Silk sericin can be cross linked, copolymerized and blended with other macromolecular materials, especially artificial polymers, to produce materials with improved properties. The protein is also used as an improving reagent or a coating material for natural and artificial fibers, fabrics and articles. The materials modified with silk sericin and sericin composites are useful as degradable biomaterials, biomedical materials and functional membranes.