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[Screening of differentially expressed proteins associated with directional highly lymphatic metastasis in ovarian
Xiao Guan1, Dan-Rong Li, Qi Wang
1Department of Gynecological Oncology, Guangxi Medical University, Nanning, Guangxi, China.
Background & Objective:
The ovarian serous papillary adenocarcinoma cell line SKOV3 and its subclones SKOV3-pm2 and SKOV3-pm3 are cell models to investigate the molecular mechanism of invasion and metastasis of ovarian cancers. This study was to screen differentially expressed proteins between ovarian carcinoma cell lines with directional (SKOV3-pm2 and SKOV3-pm3) and non-directional (SKOV3) highly lymphatic metastasis potentials using time-of-flight mass spectrometry technology and protein chips.
Methods:
The lymphatic metastasis rates of the three cell lines were detected in animal models. Proteins in endochylema and supernatants of the three cell lines were screened using surface-enhanced laser desorption and ionization-time of flight-mass spectrometry (SELDI-TOF-MS). Each sample was examined using weak cation exchange (CM-10) protein chip assay and immobilized metal affinity capture (IMAC-3) SELDI ProteinChip array. Detected protein peaks were filtrated and analyzed using Ciphergen proteinchip software 3.2.0 and Biomarker Wizard software. Differentially expressed proteins were defined as those whose absolute ratio values were greater than 0.5.
Results:
Lymphatic metastasis rates in SKOV3, SKOV3-pm2 and SKOV3-pm3 cell xenografts in nude mice were 20%, 90% and 100%, respectively (P<0.05). Proteins in endochylema whose mass-to-charge ratio (m/z) were 6971, 7475, 9089, 9453, 10103, 11655, and the protein in supernatants whose m/z was 4746 were differentially expressed in SKOV3, SKOV3-pm2 and SKOV3-pm3 cells.
Conclusion:
Combined with weak cation exchange protein chip assay and immobilized metal affinity capture SELDI ProteinChip array, SELDI-TOF-MS technology can be used to screen and identify differentially expressed proteins associated with directional highly lymphatic metastasis in ovarian carcinoma cell lines.
