人卵巢癌高低侵袭潜能细胞系差异蛋白的初步分析

The Preliminary Analysis of Differentially Expressed Proteins with High & Low Invasive Potential in Human Ovarian Cancer Cell Lines

目的:筛选与卵巢癌侵袭转移特性相关的蛋白质,为研究卵巢癌侵袭转移机制奠定基础。方法:将人卵巢癌细胞系SKOV-3在裸鼠体内多次传代,体外培养建系,结合细胞电泳速度、侵袭及移动实验筛选高、低侵袭潜能细胞亚系。采用表面增强激光解吸电离-飞行时间-质谱(SELDI-TOF-MS)检测二者的细胞裂解液,比较其蛋白质指纹图谱。结果:经过多次体内传代,结合体外实验筛选出细胞亚系SKOV-3F3,与其母系SKOV-3相比具有高侵袭潜能。比较二者蛋白质指纹图谱发现:6个差异蛋白质峰(质荷比M/Z分别为6249、14675、15425、15717、17123和17660)仅在SKOV-3中表达,3个差异蛋白质峰(M/Z分别为4355、4823和5763)仅在SKOV-3F3中表达,2个差异蛋白质峰(M/Z分别为1842和3145)均可在SKOV-3的3代亚系中捕获,但未在母系SKOV-3中发现。此外,5个差异蛋白质峰(M/Z分别为1242、5465、6899、6568和14027)在SKOV-3及其3代亚系中均可表达,但强度不同。结论:通过比较同一遗传背景的高、低侵袭潜能卵巢癌细胞的蛋白质指纹图谱,发现与侵袭潜能密切相关的差异蛋白质峰,对这些蛋白进行鉴定和功能验证有助于阐明卵巢癌侵袭转移机制。

Objective： To screen differentially expressed proteins that relate to the invasive and metastatic potential. Methods： Nude mice were intraperitoneally injected with the human ovarian cancer cell line SKOV-3. The metastatic tumors were removed and cultured in vitro. When the cell lines were established they were transplanted intraperitoneally for several cycles. The invasive potential of cell lines was evaluated by cell electrophoresis and Matrigel invasion assay. The enhanced surface laser desorption ionizing-flight time-mass spectra （ELDI-TOF-MS） was applied to detect the cell disruption solution and the protein fingerprints were compared. Results： SKOV-3F3, a cell line established after repeated in vivo passage, showed more highly invasive capacity than the parental SKOV-3 line in cell electrophoresis and Matrigel invasion assays. Based on the proteomic fingerprint patterns, we found six differelitially expressed proteins （6249, 14675, 15425, 15717, 17123 and 17660 at M/Z value） that were only expressed in SKOV-3. On the contrary, three distinct proteins （4355, 4823 and 5763 at M/Z value） were only expressed in SKOV,3F3. Furthermore, two differentially expressed proteins （1842 and 3145 at M/Z value） could only be detected in SKOV-3F1, SKOV-3F2 and SKOV-3F3. Five proteins （1242, 5465, 6899, 6568 and 14027 at M/Z value） were expressed in all SKOV-3, SKOV-3F1, SKOV-3F2 and SKOV-3F3 cell lines, but their abundance was different. Conclusion：These proteins expressed differentially between SKOV-3 and SKOV-3F3 are correlated with high invasive and metastatic potential. The identification of these proteins will help to clarify the invasion and metastasis mechanisms of ovarian cancer.