国际妇产科学杂志

国际妇产科学杂志 ›› 2018, Vol. 45 ›› Issue (6): 652-657.

• 论著 • 上一篇    下一篇

BV6诱导人卵巢癌SKOV3细胞凋亡定量蛋白组学研究

张虹，陈思思，陈琪，赵晓楠   

  1. 300100  天津市中心妇产科医院妇瘤科（张虹，陈琪）；秦皇岛市第一医院妇产科（陈思思）；滨州医学院附属医院（赵晓楠）
  • 收稿日期:2018-07-18 修回日期:2018-09-23 出版日期:2018-12-15 发布日期:2018-12-15
  • 通讯作者: 陈琪，E-mail：juseph@163.com E-mail:juseph@163.com
  • 基金资助:
    天津市卫生行业重点攻关项目（16KG113）

Quantitative Proteomics Analysis of BV6 Inducing Apoptosis in Human Ovarian Cancer SKOV3 Cells

ZHANG Hong，CHEN Si-si，CHEN Qi，ZHAO Xiao-nan   

  1. Department of Gynecology，Tianjin Central Hospital of Gynecology Obstetrics，Tianjin 300100，China（ZHANG Hong，CHEN Qi）；Department of Gynecology and Obstetrics，First Hospital of Qinhuangdao，Qinhuangdao 066000，Heibei Province，China（CHEN Si-si）；Binzhou Medical University Hospital，Binzhou 256600，Shandong Province，China（ZHAO Xiao-nan）
  • Received:2018-07-18 Revised:2018-09-23 Published:2018-12-15 Online:2018-12-15
  • Contact: CHEN Qi，E-mail：juseph@163.com E-mail:juseph@163.com

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摘要： 目的：通过定量蛋白组学技术探讨Smac类似物BV6影响人卵巢癌SKOV3细胞生长的可能机制。方法：四甲基偶氮唑蓝（MTT）法检测各浓度梯度BV6干预SKOV3细胞48 h后的生长抑制率，计算抑制率为50%时的药物浓度为半数抑制浓度（IC50），以0.05、0.1、0.2 μmol/L BV6进行后续实验；光镜下观察对照组与0.05 μmol/L BV6处理组48 h后的细胞形态变化，应用同位素标记相对和绝对定量（iTRAQ）技术结合液相串联质谱策略筛选差异蛋白并行生物信息分析；蛋白质印迹（Western blotting）检测对照组、不同浓度BV6处理组半胱氨酸天冬氨酸蛋白酶3（Caspase-3）的表达。结果：BV6可以抑制SKOV3细胞的生长增殖，且呈剂量依赖性，多组间方差分析及任意组间两两比较，差异均有统计学意义（均P＜0.05）。按公式计算BV6作用于SKOV3细胞48 h的IC50为0.40 μmol/L。基于发现错误率（FDR）＜1%，对照组与0.05 μmol/L BV6处理48 h组筛选到349个差异蛋白，其中251个蛋白表达上调、98个蛋白表达下调，经生物信息学分析显示差异蛋白与RNA转录、蛋白质翻译、细胞分裂增殖、凋亡信号调节及肿瘤血管生成密切相关。Western blotting结果显示高、中、低浓度BV6干预SKOV3细胞48 h后与对照组相比，Caspase-3蛋白表达增加，其中高BV6组的Caspase-3蛋白表达最高，组间差异有统计学意义（均P＜0.05）。结论：BV6通过抑制RNA转录、蛋白质翻译进程、细胞分裂增殖及肿瘤血管生成，促进Caspase-3凋亡信号活化介导SKOV3细胞死亡。

关键词: 卵巢肿瘤, 蛋白质组学, 细胞凋亡, 半胱氨酸天冬氨酸蛋白酶, 同位素标记

Abstract: Objective：To investigate the possible mechanism of Smac mimetic BV6 affecting the growth of human ovarian cancer SKOV3 cells by quantitative proteomics. Methods：MTT assay was used to detect the growth inhibition rate of SKOV3 cells treated with BV6 for 48 hours. Calculate the drug concentration at 50% inhibition rate as half the inhibitory concentration（IC50） and the subsequent experiments were performed at 0.05, 0.1, 0.2 μmol/L BV6. The control group and 0.05 μmol/L BV6 treatment group were observed under light microscope, after 48 h the cell morphology was changed. Application of isobarictags for relative and absolute quantitation (iTRAQ) techniques combined with Liquid Chromatography Tandem Mass Spectrometry to Screen Differential Protein Parallel Bioinformatics Analysis; Western blotting was used to detect the expression of caspase-3 in the control group and different concentrations of BV6. Results：BV6 can inhibit the proliferation of SKOV3 cells in a dose-dependent manner, and the variance analysis between groups and the comparison between any two groups were statistically significant (all P＜0.05). According to the formula, the IC50 of BV6 acting on SKOV3 cells for 48 h was 0.40 μmol/L. Based on FDR＜1%, 349 differential proteins were screened in the control group and 0.05 μmol/L BV6 for 48 h，251 were up-regulated and 98 down-regulated. Bioinformatics analysis showed differential protein and RNA transcription and protein. Translation, cell division and proliferation, apoptosis signal regulation and tumor angiogenesis were closely related. Western blotting showed that Caspase-3 protein expression was increased in the high, medium and low concentrations of BV6 for 48 h after treatment with SKOV3 cells. The expression of Caspase-3 protein was highest in the high concentration BV6 intervention group. Significance (all P＜0.05). Conclusions: BV6 promotes the death of SKOV3 cells by inhibiting RNA transcription, protein translation, cell division and proliferation, and tumor angiogenesis, and promoting the activation of Caspase-3 apoptosis.

Key words: Ovarian Neoplasms, Proteomics, Apoptosis, Caspases, Isotope labeling

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