Comprehensive Proteomic Study of the Antiproliferative Activity of a Polyphenol-Enriched Rosemary Extract on Colon Cancer Cells Using Nanoliquid Chromatography–Orbitrap MS/MS

In this work, a proteomics strategy
based on nanoliquid chromatography–tandem
mass spectrometry (nano-LC–MS/MS) using an Orbitrap high-resolution
mass spectrometer together with stable isotope dimethyl labeling (DML)
is applied to quantitatively examine relative changes in the protein
fraction of HT-29 human colon cancer cells treated with different
concentrations of a polyphenol-enriched rosemary extract over the
time. The major objective of this study was to gain insights into
the antiproliferative mechanisms induced by rosemary polyphenols.
Using this methodology, 1909 and 698 proteins were identified and
quantified in cell extracts. The polyphenol-enriched rosemary extract
treatment changed the expression of several proteins in a time- and
concentration-dependent manner. Most of the altered proteins are implicated
in the activation of Nrf2 transcription factor and the unfolded protein
response. In conclusion, rosemary polyphenols induced proteomic changes
that were related to the attenuation of aggresome formation and activation
of autophagy to alleviate cellular stress.

本研究采用基于纳升液相色谱-串联质谱（nanoliquid chromatography–tandem mass spectrometry，nano-LC–MS/MS）的蛋白质组学策略，搭配轨道阱高分辨质谱仪与稳定同位素二甲基标记（stable isotope dimethyl labeling，DML）技术，对经不同浓度富含多酚迷迭香提取物处理的HT-29人结肠癌细胞的蛋白质组分进行定量分析，以探究其随时间推移的相对变化。本研究的主要目标为深入解析迷迭香多酚诱导的抗增殖机制。通过该实验方法，研究人员在细胞提取物中共鉴定并定量得到1909种与698种蛋白质。经富含多酚的迷迭香提取物处理后，多种蛋白质的表达呈现时间与浓度依赖型变化。绝大多数表达异常的蛋白质均与Nrf2转录因子激活及未折叠蛋白质应答相关。综上，迷迭香多酚可诱导蛋白质组学改变，该改变与聚集体形成的减弱及自噬激活以缓解细胞应激密切相关。