Data on the association of CMPK1 with clinicopathological features and biological effect in human epithelial ovarian cancer

Human epithelial ovarian cancer (EOC) is the most lethal gynecological disease. However, the molecular mechanisms by which transforming growth factor-β (TGF-β) regulates ovarian tumor progression markers remain unclear. The present data show cytidine monophosphate kinase (CMPK) as an EOC biomarker and are related to the article entitled “Cytidine monophosphate kinase is inhibited by the TGF-β signalling pathway through the upregulation of miR-130b-3p in human epithelial ovarian cancer” [1]. CMPK, as well as cystatin B [2] and β-2-microglobulin [3], is overexpressed in human epithelial-type ovarian tumors. CMPK is an enzyme required for nucleic acid biosynthesis [4] and is regulated by the TGF-β signaling pathway in EOC cells [1]. Furthermore, the data show the effect of CMPK-shRNA on EOC cell apoptosis and TGF-β-induced Smad2 phosphorylation. CMPK expression in two EOC cell lines OVCAR-3 and SK-OV-3 is regulated by multiple miRNAs and some of these miRNAs may affect EOC chemoresistance [5].


Specifications
Cells were transfected with siRNA or shRNA; Cells were treated with 10 ng/ml TGF-β1 for 24 h

Experimental features
The tissue microarray included 100 paraffin-embedded ovarian tissues; Screen 9 miRNAs that potentially target CMPK1 Data source location

Shanghai, China
Data accessibility The data are with this article

Value of the data
Data present CMPK as an ovarian serous tumor progression marker. The location of CMPK protein expression in the cytoplasm and nucleus of epithelial-type ovarian tumor cells is shown.
Suppression of CMPK affects the doubling time of EOC cells. Data describe for the first time that knockdown of CMPK influences EOC cell apoptosis. Data show the effect of CMPK-shRNA on TGF-β-induced Smad2 phosphorylation.

Data
The data represent the observation from experiments of tissue microarray, Western blot and flow cytometry. Data in Table 1 are the list of sequences of siRNA, shRNA, miRNA and PCR primer used in a related research article [1]. The data of the association of CMPK protein expression with clinicopathological features of patients with epithelial ovarian tumours are shown in Table 2. Data in Fig. 1 show the positive rate for CMPK staining in the cytoplasm and nucleus.

Tissue microarray
Ovarian tissue microarray (TMA) was obtained from Xi'an Alena Biotechnology Ltd., Co. (Xi'an, Shanxi, China). Association of CMPK protein expression with clinicopathological features of patients with EOC was analyzed after immunohistochemistry staining.

Cell culture, treatment with TGF-β and transduction
OVCAR-3 and SK-OV-3 cells (ATCC, Manassas, VA, USA) were cultured in RPMI-1640 and DMEM (Corning Inc., Manassas, VA, USA), respectively. The cells were treated with 10 ng/ml of recombinant human TGF-β1 (R&D Systems, Minneapolis, MN, USA) for 24 hours. Small interfering RNA (siRNA) was purchased from GenePharma Company (Shanghai, China). The cells were transiently transfected with siRNA mixture (Roche Applied Science, Indianapolis, IN, USA) for 5 h. CMPK-shRNA was constructed with double-strand oligonucleotides. The efficiency of CMPK-shRNA lentiviral transduction was examined by fluorescence microscopy as the construct contains green fluorescent protein (GFP). Knockdown of CMPK was confirmed by qRT-PCR and Western blot. The target position in CMPK mRNA sequence (GenBank Accession: NM_016308) and β-actin mRNA sequence (GenBank Accession: NM_001101) is shown. Sequence in lowercase indicates a linker. NC, negative control; nt, nucleotide; PCR, polymerase chain reaction; siRNA, small interfering RNA; shRNA, short hairpin RNA.

Transfection of miRNA mimics
Nine miRNA mimics and negative control miRNA were purchased from Guangzhou RiboBio Co., Ltd. (Guangzhou, Guangdong, China). The cells were transfected with miRNA mimics for 5 h and then incubated in a complete medium for up to 72 hours.

Flow cytometry
Apoptotic cells were detected by staining cells with APC Annexin-V and propidium iodide (PI) using an Annexin-V Apoptosis Detection Kit (BD Pharminggen, San Diego, CA, USA). Lentivirus infected EOCs were seeded in a 6-well plate and were incubated for 24 hours. Both supernatant and attached cells were collected and resuspended in 100 ml of 1X binding buffer. After adding 5 ml of Annexin-V and/or 5 ml of PI, the cells were incubated in the dark at room temperature for 15 min. After adding 400 ml of 1X binding buffer, the cell population was analyzed by flow cytometry. Fig. 6. Effect of miRNAs on CMPK expression. CMPK protein expression was detected by Western blot. OVCAR-3 and SK-OV-3 cells were transiently transfected with nine miRNA mimics. Negative control miRNA (miR-NC) was used as control. Experiment was repeated twice.