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Published on: November 18, 2019
iTRAQ-based quantitative proteomic analysis of Yamanaka factors reprogrammed breast cancer cells
Kun Wang1, Zhiyan Shan1,2, Lian Duan1,3
1Department of Histology and Embryology, Harbin Medical University, Harbin, China.
Abstract:
Cancer cells had been developed to be reprogrammed into embryonic stem like cells by induced pluripotent stem cells (iPSCs) technology, however, the tumor differentiation/dedifferentiation mechanisms had not yet been analyzed on a genome-wide scale. Here, we inserted the four stem cell transcription factor genes OCT4, SOX2, C-MYC and KLF4 into MCF cells (MCFs), represented a female breast cancer cell type, and obtained iPSCs (Mcfips) in about 3 weeks. By using the LC MS/MS iTRAQ technology, we analyzed the proteomic changes between MCFs and Mcfips. Of identified 4,616 proteins totally, 247 and 142 differentially expressed (DE) proteins were found in Mcfips compared with human induce pluripotent stem cells (Hips) and MCFs, respectively. 35 co-up and 10 co-down regulated proteins were recognized in DE proteins. Above DE proteins were categorized with GO functional classification annotation and KEGG metabolic pathway analysis into biological processes. In the protein interaction network, we found 37 and 39 hubs interacted with more than one protein in Mcfips comparing to Hips, in addition, 25 and 9 hubs were identified in Mcfips comparing to MCFs. Importantly, the mitochondria, ribosome and tumor suppressor proteins were found to be core regulators of tumor reprogramming, which might contribute to understand the mechanisms in relation to the occurrences and progression of a tumor. Thus, our study provided a valuable data for exploring the possibility to normalize the malignant phenotype.
Insights
Induced pluripotent stem cells (iPSCs) technology reprogrammed cancer cells. This study analyzed genome-wide proteomic changes, identifying mitochondria, ribosome, and tumor suppressor proteins as key regulators in tumor reprogramming.
Area of Science:
- Biochemistry
- Molecular Biology
- Cancer Research
Background:
- Induced pluripotent stem cells (iPSCs) technology offers a method to reprogram cancer cells into a stem-like state.
- Understanding the genome-wide mechanisms of tumor differentiation and dedifferentiation is crucial for cancer research.
Purpose of the Study:
- To analyze the proteomic changes during the reprogramming of breast cancer cells (MCFs) into induced pluripotent stem cells (Mcfips).
- To identify key proteins and pathways involved in tumor reprogramming using a genome-wide approach.
Main Methods:
- Reprogramming of MCF cells using OCT4, SOX2, C-MYC, and KLF4 transcription factors to generate Mcfips.
- Proteomic analysis using LC-MS/MS iTRAQ technology to compare protein expression between MCFs, Mcfips, and human induced pluripotent stem cells (Hips).
- Gene Ontology (GO) functional classification and KEGG pathway analysis for differentially expressed proteins and protein interaction network analysis.
Main Results:
- Identified 4,616 proteins, with 247 differentially expressed in Mcfips compared to Hips and 142 compared to MCFs.
- Discovered 35 co-upregulated and 10 co-downregulated proteins.
- Identified key protein regulators including mitochondria, ribosome, and tumor suppressor proteins involved in tumor reprogramming.
Conclusions:
- Mitochondria, ribosome, and tumor suppressor proteins are core regulators of tumor reprogramming.
- This study provides valuable proteomic data for understanding tumor reprogramming mechanisms.
- Findings may contribute to exploring strategies for normalizing malignant phenotypes in cancer cells.
