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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 the proteins related to the invasion and metastasis of ovarian cancer and lay a foundation for studying the mechanism of invasion and metastasis of ovarian cancer. Methods: Human ovarian cancer cell line SKOV-3 was sub-cultured in nude mice for many times and cultured in vitro. The high and low invasive potential cell lines were screened according to the cell electrophoresis speed, invasion and migration experiments. The cell lysates of both were detected by surface-enhanced laser desorption ionization-time of flight mass spectrometry (SELDI-TOF-MS), and their protein fingerprints were compared. RESULTS: After many passages in vivo and in vitro, SKOV-3F3 cell line was screened out and had high invasive potential compared with its parental line SKOV-3. Comparison of the protein fingerprints of the two revealed that the six differential protein peaks (mass-to-charge ratio M / Z 6249,14675,15425,15717,17123 and 17660 respectively) were only expressed in SKOV-3, and the three differential protein peaks (M / Z were 4355, 4823 and 5763, respectively) were only expressed in SKOV-3F3. Two differential protein peaks (M / Z 1842 and 3145, respectively) were captured in the third generation of SKOV-3 but not in maternal Found in SKOV-3. In addition, five different protein peaks (M / Z 1242, 5465, 6899, 6568 and 14027, respectively) were expressed in SKOV-3 and its 3rd generation sublines, but with different intensities. Conclusion: By comparing the protein fingerprints of high and low invasion potential ovarian cancer cells with the same genetic background, we found that the differential protein peaks are closely related to invasion potential. Identification and functional verification of these proteins may help elucidate the mechanism of invasion and metastasis of ovarian cancer.