掌叶半夏总蛋白作用人卵巢癌SKOV3细胞的蛋白质组学研究

作者	单位
周莉	温州医学院附属第一医院妇产科
郑飞云	温州医学院附属第一医院妇产科
王汉楚	温州医学院附属第一医院妇产科
陈小燕	温州医学院附属第一医院妇产科
包其郁	温州医学院生命科学院

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中文摘要:

目的研究掌叶半夏总蛋白作用于人卵巢癌SKOV3细胞后蛋白质组学的变化。方法体外培养SKOV3细胞,应用CCK-8比色法观察不同时间(24、48、72、96 h)、不同浓度(0、0.05、0.10、0.20、0.30、0.40、0.50 mg/mL)掌叶半夏总蛋白对SKOV3细胞的生长抑制作用。取对数生长期SKOV3细胞,加入0.296 mg/mL掌叶半夏总蛋白作用于细胞48 h,提取细胞总蛋白,采用双向凝胶电泳(2-DE)和基质辅助激光解吸电离飞行时间质谱(MALDI-TOF-MS)筛选差异表达的蛋白质。结果 0.10～0.50 mg/mL掌叶半夏总蛋白可显著抑制SKOV3细胞的生长,且具有一定的时效和量效关系(P<0.05,P<0.01)。经2-DE和质谱分析成功鉴定43个差异蛋白质点(其中21个上调,22个下调),包括α-烯醇化酶1、真核翻译起始因子3α、亲环素B等。结论掌叶半夏总蛋白可显著抑制SKOV3细胞的生长,并引起SKOV3细胞株蛋白质组学的改变,可能与其抗癌机制相关。

英文摘要:

Objective To study the proteomics of Pinellia pedatisecta Schott total protein on human ovarian cancer SKOV3 cells.Methods SKOV3 cells were in vitro cultured.The growth inhibition of SKOV3 cells in the logarithmic phase with different concentrations of Pinellia pedatisecta Schott(0,0.05,0.10,0.20,0.30,0.40,and 0.50 mg/mL) at different time points(24,48,72,and 96 h) was analyzed by CCK-8 colorimetry.The total protein was extracted after adding 0.296 mg/mL Pinellia pedatisecta Schott protein for 48 h.The protein with differential expressions was screened out using two-dimensional gel electrophoresis(2-DE) and matrix-assisted laser desorption ionization time of flight mass spectrometry(MALDI-TOF-MS).Results The protein of Pinellia pedatisecta Schott(at 0.10-0.50 mg/mL) could significantly inhibit the growth of SKOV3 cells in a time-and dose-dependent manner(P<0.05,P<0.01).After analyzed by 2-DE and MALDI-TOF-MS,43 differential protein dots were successfully identified(21 up-regulated and 22 down-regulated),including alpha-enolase 1,eukaryotic initiation factor 3α,cyclophilin B,and so on.Conclusions Protein of Pinellia pedatisecta Schott could significantly inhibit the growth of SKOV3 cells,and lead to the proteomics changes of SKOV3 cell strain.They might be correlated with its anti-tumor mechanisms.

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