Identification of COPA as a potential prognostic biomarker and pharmacological intervention target of cervical cancer by quantitative proteomics and experimental verification.

BACKGROUND: Cervical cancer is the most fatal gynecological carcinoma in the world. It is urgent to explore novel prognostic biomarkers and intervention targets for cervical cancer. METHODS: Through integrated quantitative proteomic strategy, we investigated the protein expression profiles of cervical cancer; 28 fresh frozen tissue samples (11 adenocarcinoma (AC), 12 squamous cell carcinoma (SCC) and 5 normal cervixes (HC)) were included in discover cohort; 45 fresh frozen tissue samples (19 AC, 18 SCC and 8 HC) were included in verification cohort; 140 paraffin-embedded tissues samples of cervical cancer (85 AC and 55 SCC) were used for immunohistochemical evaluation (IHC) of coatomer protein subunit alpha (COPA) as a prognostic biomarker for cervical cancer; how deficiency of COPA affects cell viability and tumorigenic ability of cervical cancer cells (SiHa cells and HeLa cells) were evaluated by cell counting kit-8 and clone formation in vitro. RESULTS: We identified COPA is a potential prognostic biomarker for cervical cancer in quantitative proteomics analysis. By retrospective IHC analysis, we additionally verified the proteomics results and demonstrated moderate or strong IHC staining for COPA is an unfavourable independent prognostic factor for cervical cancer. We also identified COPA is a potential pharmacological intervention target of cervical cancer by a series of in vitro experiments. CONCLUSION: This study is the first to demonstrate that COPA may contribute to progression of cervical cancer. It can serve as a potential prognostic biomarker and promising intervention target for cervical cancer.

Activin A induces tumorigenesis of leiomyoma via regulation of p38ß MAPK-mediated signal cascade.

OBJECTIVE: The objective of this study was to investigate the role of p38-C/EBPß signaling in leiomyoma cells and myometrial cells challenged with Activin A, and to identify specifically the isoform of p38 MAPK that mediates the effects of Activin A. METHODS: The immortalization human leiomyoma cells (HuLM) and human myometrial cells (HM), and mouse myometrial tissues were treated with Activin A (4 nM) in response to p38α/ß inhibition (10 µM SB202190) or depletion (p38 α/ß-targeting siRNA or p38ß muscle specific-knock out mice). p38 MAPK signaling molecules (p-p38 and p-C/EBPß) and ECM components (COL1A1 and/or FN) were analyzed by Western blotting. RESULTS: Activin A induced ECM accumulation in leiomyoma cells and myofibroblastic transformation in myometrical cells specifically by p38ß MAPK. CONCLUSION: This study is the first to demonstrate that activation of C/EBPß by p38ß MAPK may contribute to tumorigenesis and progression of Activin A-induced leiomyoma. Specific p38ß inhibition may represent a novel and promising intervention for leiomyoma.

Activin A induces leiomyoma cell proliferation, extracellular matrix (ECM) accumulation and myofibroblastic transformation of myometrial cells via p38 MAPK.

OBJECTIVE: The objective of this study was to evaluate the role of Activin A and p38ß MAPK-activated signaling in human leiomyoma cells, myometrial cells and mouse myometrial tissues. METHODS: The immortalization human leiomyoma cells (HuLM), the immortalized human myometrial cells (HM) and mouse myometrial tissues were treated with Activin A (4 nM) and/or specific p38 inhibitor SB202190 (10 µM) for different days of interval (to measure proliferation rate) or 1 h (to measure signaling molecules) or 48 h (to measure proliferating markers Ki-67, ECM mRNA, and/or ECM protein expression) by real-time PCR, Western blot, and/or immunocytochemistry. RESULTS: Activin A induced cell proliferation and ECM proteins accumulation in HuLM cells via p38 MAPK. Activin A also induced myofibroblastic transformation in HM cells and mouse myometrical tissues via the phosphorylation of p38. The effects of Activin A in leiomyoma cells, myometrical cells and tissues were abolished by p38α/ß MAPK inhibitor SB202190. CONCLUSION: This study demonstrates Activin A-p38 MAPK signaling pathway in leiomyoma and myometrium may contribute to excessive ECM production, leiomyoma growth and progression. Targeting Activin A-p38 MAPK signaling pathway could be a potential therapeutic intervention for uterine leiomyoma.

Efficient Photoacoustic Imaging With Biomimetic Mesoporous Silica-Based Nanoparticles.

Indocyanine green (ICG), a near-infrared (NIR) fluorescent dye approved by the Food and Drug Administration (FDA), has been extensively used as a photoacoustic (PA) probe for PA imaging. However, its practical application is limited by poor photostability in water, rapid body clearance, and non-specificity. Herein, we fabricated a novel biomimetic nanoprobe by coating ICG-loaded mesoporous silica nanoparticles with the cancer cell membrane (namely, CMI) for PA imaging. This probe exhibited good dispersion, large loading efficiency, good biocompatibility, and homologous targeting ability to Hela cells in vitro. Furthermore, the in vivo and ex vivo PA imaging on Hela tumor-bearing nude mice demonstrated that CMI could accumulate in tumor tissue and display a superior PA imaging efficacy compared with free ICG. All these results demonstrated that CMI might be a promising contrast agent for PA imaging of cervical carcinoma.

p16INK4A and Ki-67 immunostaining on cell blocks from residual ThinPrep material is helpful in identifying significant preneoplastic cervical lesions.

Cell blocks can be prepared from residual ThinPrep material, and immunohistochemical staining can be used. The objectives of the current study were (1) to investigate the role of cell block preparation in identifying significant preneoplastic cervical lesions; and (2) to assess the diagnostic value of p16(INK4A) and Ki-67 immunostaining on cell blocks to identify significant preneoplastic cervical lesions. Samples from residual substances of ThinPrep from 79 patients were collected for cell block preparations. Cell block sections and histological sections of the same patients were stained with hematoxylin and eosin, and were immunostained with antibodies against p16(INK4A) and Ki-67 antigen. The sensitivity and specificity for p16(INK4A) and Ki-67 immunostaining to detect high-grade squamous intraepithelial neoplasia were 95.12% and 73.68%, respectively. The positive predictive value and negative predictive value were 79.59% and 93.33%, respectively. Immunostaining of cell blocks for p16(INK4A) and Ki-67 exhibited a statistically significant association with the presence of significant lesions on cell blocks (P < 0.05). p16(INK4A) and Ki-67 immunostaining on cell blocks from residual ThinPrep material is helpful in identifying significant preneoplastic cervical lesions.