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Comparative proteomics analysis of the proteins associated with laryngeal carcinoma-related gene 1
Xiaopeng Zhang1, Zhiqiang Xiao, Zhuchu Chen
1From Key Laboratory of Cancer Proteomics, Ministry of Health of China 'Xiangya Hospital, and the Cancer Research Institute, Central South University, Changsha, China.
Objectives:
A novel gene, laryngeal carcinoma-related gene 1 (LCRG1), had the characteristics of tumor-suppressor genes. It was cloned in our laboratory. The objective was to find and characterize the proteins related to LCRG1 and to elucidate the molecular mechanism of LCRG1.
Study Design:
We used the established cell lines of Hep-2/LCRG1 (Hep-2 cells transfected by recombinant plasmid pcDNA3.1[+]/LCRG1) and Hep-2/pcDNA3.1(+) (Hep-2 cells transfected by control vector pcDNA3.1[+]) as cell models.
Methods:
Two-dimensional gel electrophoresis (2-DE) technology was performed to separate the proteins of Hep-2/LCRG1 and Hep-2/pcDNA3.1(+) cell lines, respectively. The differential protein spots were analyzed by software analysis, subject to in-gel digestion, and identified by Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) and electrospray ionization-quadruple time-of-flight MS/MS (ESI-Q-TOF MS/MS). Then the differential expression levels of partial identified proteins were determined by Western blotting analysis and quantitative real-time reverse transcriptase-polymerase chain reaction.
Results:
The results showed the attained 2-DE patterns of the two cell lines were well-resolved and reproducible. There were 1075+/-43 and 1027+/-23 protein spots observed in Hep-2/LCRG1 and Hep-2/pcDNA3.1(+) cell lines, respectively. The average matching rate of the two cell lines was 91%. Twenty-six differentially expressed protein spots were identified (twenty spots for MALDI-TOF-MS, six spots for ESI-Q-TOF MS/MS). Most of the characterized proteins were characterized as the members of enzymes (phosphoglycerate mutase, manganese superoxide dismutase, and so on), transcription proteins (rho gdp dissociation inhibitor), and so on. Those identified proteins might contribute to the tumor-suppressive function of LCRG1. The differential expression levels of the partial proteins were confirmed by real-time polymerase chain reaction and Western blotting.
Conclusions:
We tentatively proposed those differentially expressed proteins were involved in the tumor-suppressive process related to LCRG1. These data will be helpful to elucidate the molecular mechanism of LCRG1.
Insights
This study identified proteins related to laryngeal carcinoma-related gene 1 (LCRG1), a tumor suppressor. These proteins are involved in LCRG1's cancer-fighting function, aiding in understanding its molecular mechanism.
Area of Science:
- Molecular biology
- Cancer research
- Proteomics
Background:
- Laryngeal carcinoma-related gene 1 (LCRG1) exhibits tumor-suppressor characteristics.
- Understanding the molecular mechanisms of LCRG1 is crucial for cancer research.
Purpose of the Study:
- To identify and characterize proteins associated with LCRG1.
- To elucidate the molecular mechanism underlying LCRG1's tumor-suppressive function.
Main Methods:
- Utilized Hep-2 cell lines stably transfected with LCRG1 or a control vector.
- Employed two-dimensional gel electrophoresis (2-DE) for protein separation.
- Identified differentially expressed proteins using MALDI-TOF-MS and ESI-Q-TOF MS/MS, with validation by Western blotting and qRT-PCR.
Main Results:
- Successfully resolved and reproduced 2-DE patterns, identifying 26 differentially expressed protein spots.
- Key identified proteins include enzymes (e.g., phosphoglycerate mutase, manganese superoxide dismutase) and transcription factors (e.g., rho gdp dissociation inhibitor).
- Differential expression levels were confirmed, suggesting these proteins contribute to LCRG1's tumor-suppressive activity.
Conclusions:
- The identified differentially expressed proteins are tentatively proposed to be involved in the tumor-suppressive process mediated by LCRG1.
- These findings provide valuable insights into the molecular mechanism of LCRG1 in cancer suppression.
