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Published on: July 30, 2018
Proteomic analysis of cervical cancer cells treated with adenovirus-mediated MDA-7
Lili Wei1, Zhongtang Wang, Tao Cui
1Department of Biochemistry and Molecular Biology, Chongqing University of Medical Sciences, Shandong Cancer Hospital, China.
Abstract:
Ad.mda-7 inhibited growth and decreased survival in a broad array of human tumor cells, without eliciting detrimental effects in normal cells. This study demonstrates that Ad.mda-7 can effectively impede the proliferation and induce apoptosis of human cervical carcinoma cells, but the underlying mechanisms inducing cell death at protein level are unknown. Using proteome analysis, an investigation aimed at a better understanding of the antiproliferative mechanisms by Ad.mda-7 was carried out in CaSki cervical cancer cells. A total of 43 differentially expressed proteins were visualized by 2-DE and silver stain., 29 proteins of which were identified via matrix-assisted laser desorption/ionization-time of flight mass spectrometry(MALDI-TOF-MS) analysis, 15 were upregulated (eg., Tumor suppressor p53, Apoptosis regulator BAX, Adenylate kinase isoenzyme 1(AK1), Growth arrest and DNA-damage-inducible protein GADD45 gamma (GADD45gamma)) and 14 were downregulated (e.g., Eukaryotic translation initiation factor 5A(eIF-5A), Protein DJ-1, Annexin V, Transcription elongation factor B polypeptide 2 (TCEB2), TRAF family member-associated NFkappaB activator (TRAF2),c-Myc-responsive protein Rcl (RCL)). Among the identified proteins, the protein and mRNA alterations of six proteins were further confirmed by Western blot and semi-quantitative RT-PCR. Together, at both the mRNA and protein levels, p53, BAX, AK1, GADD45gamma and BCCIP were upregulated, while eIF-5A was downregulated following Ad.mda-7 treatment. Our findings may offer new insights into the antiproliferative mechanisms by Ad.mda-7 and its mode of action in cervical carcinoma cells.
Insights
Ad.mda-7 inhibits cervical cancer cell growth by altering protein expression. This study identified key upregulated proteins like p53 and BAX, and downregulated proteins such as eIF-5A, revealing Ad.mda-7
Area of Science:
- Molecular Biology
- Oncology
- Biochemistry
Background:
- Ad.mda-7 demonstrates broad-spectrum anti-tumor activity without harming normal cells.
- The precise protein-level mechanisms of Ad.mda-7-induced apoptosis in cervical cancer remain unclear.
Purpose of the Study:
- To elucidate the antiproliferative mechanisms of Ad.mda-7 in cervical carcinoma cells.
- To identify key protein expression changes induced by Ad.mda-7 in CaSki cells using proteome analysis.
Main Methods:
- Proteome analysis using 2-DE and silver staining to visualize differentially expressed proteins.
- Identification of proteins using MALDI-TOF-MS.
- Validation of protein and mRNA alterations via Western blot and semi-quantitative RT-PCR.
Main Results:
- 43 differentially expressed proteins were identified in CaSki cells treated with Ad.mda-7.
- 15 proteins, including p53, BAX, AK1, and GADD45gamma, were upregulated.
- 14 proteins, including eIF-5A, Protein DJ-1, and Annexin V, were downregulated.
- Upregulation of p53, BAX, AK1, GADD45gamma, and BCCIP, and downregulation of eIF-5A were confirmed at both mRNA and protein levels.
Conclusions:
- Ad.mda-7 treatment leads to significant alterations in protein expression profiles in cervical cancer cells.
- The identified protein changes provide insights into the molecular mechanisms underlying Ad.mda-7's anti-proliferative and apoptosis-inducing effects.
- These findings contribute to understanding Ad.mda-7's mode of action in cervical carcinoma.
