Journal: Scientific Reports

Article Title: Proteomics analysis reveals FEN1 as a promising therapeutic target against small cell neuroendocrine carcinoma of the cervix

doi: 10.1038/s41598-025-12892-w

Figure Lengend Snippet: The expression of FEN1 and the regulation of FEN1 on PCNA in SCNECC. A Immunohistochemical detection and expression analysis of FEN1 in SCNECC. B Expression levels were scored and analyzed, with statistical significance indicated by asterisks compared to adjacent non-cancerous tissues. C The interaction network of FEN1 was obtained from the STRING database. D TC-YIK cells were transfected with either negative control (NC) siRNA or FEN1 -specific siRNA for 24 h. Relative mRNA expression levels of FEN1 and PCNA were detected by qRT-PCR, with GAPDH serving as the internal reference gene. n = 3. E Following transfection of TC-YIK cells with NC or FEN1 siRNA for 48 h, protein expression levels of FEN1 and PCNA were examined by WB analysis. Grayscale analysis and statistical evaluation of the WB results were performed using ImageJ software. Two-independent-samples t-tests was employed to analyze the experimental data.

Article Snippet: The membranes were blocked for 1 h at room temperature in 5% (w/v) nonfat dried milk and then incubated with primary antibodies specific for FEN1 (Catalog Number: 14768-1-AP; 1:2000, Proteintech Group, China), PCNA (Catalog Number: 10205-2-AP; 1:10000, Proteintech Group, China), Caspase-9 (Catalog Number: 10380-1-AP; 1:300, Proteintech, China), BCL-2 (Catalog Number: 12789-1-AP; 1:9000, Proteintech Group, China), PIK3CA (Catalog Number: PB0351; 1:1000, Biological Technology Co., Ltd., China), and GAPDH (Catalog Number: AP0063, Bioworld Technology, Inc., China).

Techniques: Expressing, Immunohistochemical staining, Transfection, Negative Control, Quantitative RT-PCR, Software

Journal: Scientific Reports

Article Title: Proteomics analysis reveals FEN1 as a promising therapeutic target against small cell neuroendocrine carcinoma of the cervix

doi: 10.1038/s41598-025-12892-w

Figure Lengend Snippet: The role of FEN1/PCNA in TC-YIK Cells. ( A ) After transfecting TC-YIK cells with NC, FEN1 siRNA or PCNA siRNA, cell proliferation activity was assessed at 0 h, 24 h, 48 h and 72 h using the CCK-8 method, and statistical analysis was conducted. n = 5. ANOVA was employed to analyze the experimental data. ( B ) Apoptosis was detected using flow cytometry 48 h post-transfection, and the results were statistically analyzed. n = 3. ( C ) Colony formation assay and quantitative analysis of colony numbers using crystal violet staining, n = 3. ( D ) Cell cycle changes were analyzed by flow cytometry 48 h after transfection, and statistical analysis of the results was performed. n = 3. ( E ) After transfecting TC-YIK cells with NC or FEN1 siRNA for 24 h, qRT-PCR was performed to determine the relative mRNA expression levels of BCL-2 , PIK3CA , and Caspase-9 , using GAPDH as the internal reference gene. n = 3. ( F ) WB analysis was conducted to examine the protein expression levels of BCL-2, PIK3CA, and Caspase-9 in TC-YIK cells 48 h after transfection with NC or FEN1 siRNA. Grayscale analysis and statistical evaluation of the WB bands from Panel F were performed using ImageJ software. Data are presented as means ± SDs. Comparisons with the NC group: *: P < 0.05; **: P < 0.01. Two-independent-samples t-tests was employed to analyze the experimental data in B-F.

Article Snippet: The membranes were blocked for 1 h at room temperature in 5% (w/v) nonfat dried milk and then incubated with primary antibodies specific for FEN1 (Catalog Number: 14768-1-AP; 1:2000, Proteintech Group, China), PCNA (Catalog Number: 10205-2-AP; 1:10000, Proteintech Group, China), Caspase-9 (Catalog Number: 10380-1-AP; 1:300, Proteintech, China), BCL-2 (Catalog Number: 12789-1-AP; 1:9000, Proteintech Group, China), PIK3CA (Catalog Number: PB0351; 1:1000, Biological Technology Co., Ltd., China), and GAPDH (Catalog Number: AP0063, Bioworld Technology, Inc., China).

Techniques: Activity Assay, CCK-8 Assay, Flow Cytometry, Transfection, Colony Assay, Staining, Quantitative RT-PCR, Expressing, Software

Journal: Scientific Reports

Article Title: Proteomics analysis reveals FEN1 as a promising therapeutic target against small cell neuroendocrine carcinoma of the cervix

doi: 10.1038/s41598-025-12892-w

Figure Lengend Snippet: In Vivo Assessment of the Effect of FEN1 Knockdown on Tumor Growth in SCNECC Xenografts. A Representative images of tumor-bearing nude mice from the in vivo NC siRNA group and the in vivo FEN1 siRNA group. B Comparison of body weights of nude mice with tumors in the in vivo NC group and the in vivo FEN1 siRNA group. C Excised tumor samples from nude mice in the in vivo NC group and the in vivo FEN1 siRNA group. D Comparison of tumor weights from nude mice in the in vivo NC group and the in vivo FEN1 siRNA group. E Comparison of tumor volumes in nude mice from the in vivo NC group and the in vivo FEN1 siRNA group. ANOVA was employed to analyze the experimental data. F qRT-PCR was performed to detect the relative mRNA expression levels of FEN1 , PCNA , BCL-2 , PIK3CA , and Caspase-9 in the SCNECC xenografts, using GAPDH as the internal reference gene. n = 3. G The relative protein expression levels of FEN1, PCNA, BCL-2, PIK3CA, and Caspase-9 were examined by WB analysis in the SCNECC xenografts, using GAPDH as the internal reference gene. n = 3. Grayscale analysis and statistical evaluation of the WB bands were performed using ImageJ software. Compared to the in vivo NC group, ns: P > 0.05 indicates no statistical significance, *: P < 0.05 indicates statistical significance, while **: P < 0.01 indicates high statistical significance. Two-independent-samples t-tests was employed to analyze the experimental data in the B, D, F, and G.

Article Snippet: The membranes were blocked for 1 h at room temperature in 5% (w/v) nonfat dried milk and then incubated with primary antibodies specific for FEN1 (Catalog Number: 14768-1-AP; 1:2000, Proteintech Group, China), PCNA (Catalog Number: 10205-2-AP; 1:10000, Proteintech Group, China), Caspase-9 (Catalog Number: 10380-1-AP; 1:300, Proteintech, China), BCL-2 (Catalog Number: 12789-1-AP; 1:9000, Proteintech Group, China), PIK3CA (Catalog Number: PB0351; 1:1000, Biological Technology Co., Ltd., China), and GAPDH (Catalog Number: AP0063, Bioworld Technology, Inc., China).

Techniques: In Vivo, Knockdown, Comparison, Quantitative RT-PCR, Expressing, Software

Journal: Scientific Reports

Article Title: Proteomics analysis reveals FEN1 as a promising therapeutic target against small cell neuroendocrine carcinoma of the cervix

doi: 10.1038/s41598-025-12892-w

Figure Lengend Snippet: Effects of FEN1 Inhibitor SC13 on Clonogenic Capacity, Proliferation, Apoptosis, and Cell Cycle of TC-YIK Cells. After treatment with the SC13 inhibitor in TC-YIK cells, the following assessments were performed: ( A ) Cell proliferation activity was measured using the CCK-8 assay at 0 h, 24 h, 48 h and 72 h, followed by statistical analysis. n = 5. ANOVA was employed to analyze the experimental data. ( B ) Flow cytometry was used to detect apoptosis 48 h after SC13 inhibitor treatment, with subsequent statistical analysis. n = 3. ( C ) Colony formation was visualized by crystal violet staining, and the number of colonies was statistically analyzed. n = 3. ( D ) Flow cytometry was employed to analyze changes in the cell cycle 48 h after SC13 inhibitor treatment, followed by statistical analysis. n = 3. ( E ) Relative mRNA expression levels of FEN1 , PCNA , BCL-2 , PIK3CA , and Caspase-9 were measured by qRT-PCR 24 h after SC13 inhibitor treatment, using GAPDH as the internal reference gene. n = 3. ( F ) WB analysis was performed to examine the protein expression levels of FEN1, PCNA, BCL-2, PIK3CA, and Caspase-9 48 h after SC13 inhibitor treatment. Grayscale analysis and statistical evaluation of the WB bands from Panel G were conducted using ImageJ software. n = 3. Comparisons with the NC group: *: P < 0.05 indicates statistical significance, while **: P < 0.01 indicates high statistical significance. Two-independent-samples t-tests was employed to analyze the experimental data in the B-F.

Article Snippet: The membranes were blocked for 1 h at room temperature in 5% (w/v) nonfat dried milk and then incubated with primary antibodies specific for FEN1 (Catalog Number: 14768-1-AP; 1:2000, Proteintech Group, China), PCNA (Catalog Number: 10205-2-AP; 1:10000, Proteintech Group, China), Caspase-9 (Catalog Number: 10380-1-AP; 1:300, Proteintech, China), BCL-2 (Catalog Number: 12789-1-AP; 1:9000, Proteintech Group, China), PIK3CA (Catalog Number: PB0351; 1:1000, Biological Technology Co., Ltd., China), and GAPDH (Catalog Number: AP0063, Bioworld Technology, Inc., China).

Techniques: Activity Assay, CCK-8 Assay, Flow Cytometry, Staining, Expressing, Quantitative RT-PCR, Software