znf695  (Proteintech)

Journal: Oncology Reports

Article Title: Zinc finger protein 695 facilitates the proliferation of colorectal cancer cells through activation of the NEK2 and PI3K/Akt/mTOR signaling pathways

doi: 10.3892/or.2025.8949

Figure Lengend Snippet: Clinical relevance of ZNF695 in patients with CRC. (A) Transcript levels of ZNF695 in pan-cancer was analyzed using UALCAN based on TCGA data. *P<0.05 vs. normal group. (B) Transcript levels of ZNF695 in cancer (n=275) and non-cancer (n=349) tissues of patients with CRC. *P<0.05. (C) mRNA expression of ZNF695 in CRC tissues based on the individual cancer stage and in non-cancer tissues was analyzed using Gene Expression Profiling Interactive Analysis. ***P<0.001. (D) Promoter methylation level of ZNF695 in cancer (n=313) and non-cancer (n=37) tissues of patients with CRC was analyzed using UALCAN. P>0.05. (E) Promoter methylation level of ZNF695 in in CRC tissues based on the individual cancer stage and in non-cancer tissues was analyzed using UALCAN. Normal vs. stage 2, P<0.05. (F) Overall survival of patients with CRC who were divided into the ZNF695 high expression group (n=406) and the low expression group (n=655) was analyzed from Kaplan Meier plotter. P=0.065. (G) mRNA expression levels of ZNF695 were detected by reverse transcription-quantitative PCR in NCM460, HT-29, HCT-116 and HCT-8 cells. *P<0.05, **P<0.01 vs. NCM460. BLCA, bladder urothelial carcinoma; BRCA, breast invasive carcinoma; CESC, cervical squamous cell carcinoma and endocervical adenocarcinoma; CHOL, cholangiocarcinoma; CRC, colorectal cancer; ESCA, esophageal carcinoma; GBM, glioblastoma; HNSC, head and neck squamous cell carcinoma; KICH, kidney chromophobe; KIRC, kidney renal clear cell carcinoma; KIRP, kidney renal papillary cell carcinoma; LIHC, liver hepatocellular carcinoma; LUAD, lung adenocarcinoma; LUSC, lung squamous cell carcinoma; PAAD, pancreatic adenocarcinoma; PRAD, prostate adenocarcinoma; PCPG, pheochromocytoma and paraganglioma; READ, rectum adenocarcinoma; SARC, sarcoma; SKCM, skin cutaneous melanoma; THCA, thyroid carcinoma; THYM, thymoma; STAD, stomach adenocarcinoma; UCEC, uterine corpus endometrial carcinoma; TCGA, The Cancer Genome Atlas; ZNF695, zinc finger protein 695.

Article Snippet: The following primary antibodies were used: ZNF695 (cat. no. 25556-1-AP; 1:800; Proteintech Group, Inc.), NEK2 (cat. no. 24171-1-AP; 1:800; Proteintech Group Inc.), Akt (cat. no. 9272; 1:1,000; Cell Signaling Technology, Inc.), phosphorylated (p)-Akt (cat. no. 4060; 1:1,000; Cell Signaling Technology, Inc.), ribosomal protein S6 (S6; cat. no. 2217; 1:1,000; Cell Signaling Technology, Inc.), p-S6 (cat. no. 2211; 1:1,000; Cell Signaling Technology, Inc.), p21 (cat. no. 2947; 1:1,000; Cell Signaling Technology, Inc.), Bcl-2 (cat. no. 3498; 1:1,000; Cell Signaling Technology, Inc.), β-actin (cat. no. 66009-1-Ig; 1:5,000; Proteintech Group Inc.) and GAPDH (cat. no. 60004-1-Ig; 1:5,000; Proteintech Group Inc.).

Techniques: Expressing, Gene Expression, Methylation, Reverse Transcription, Real-time Polymerase Chain Reaction

Journal: Oncology Reports

Article Title: Zinc finger protein 695 facilitates the proliferation of colorectal cancer cells through activation of the NEK2 and PI3K/Akt/mTOR signaling pathways

doi: 10.3892/or.2025.8949

Figure Lengend Snippet: Overexpression of ZNF695 supports the proliferation of CRC cells. (A) mRNA and protein levels of ZNF695 in siCtrl, siZNF695#1 and siZNF695#2 HCT-8 cells were detected using RT-qPCR and western blotting, respectively. **P<0.01 vs. siCtrl group. (B) Proliferation of HCT-8 cells transfected with siCtrl, siZNF695#1 and siZNF695#2 was determined using a CCK8 assay. *P<0.05, **P<0.01 vs. siCtrl group. (C) mRNA and protein levels of ZNF695 in Ctrl and ZNF695-overexpressing HT-29 cells were detected using RT-qPCR and western blotting, respectively. **P<0.01 vs. Ctrl group. (D) Proliferation of Ctrl and ZNF695-overexpressing HT-29 cells was determined by CCK8 assay. *P<0.05, **P<0.01 vs. Ctrl group. Colony formation was assessed in (E) siCtrl, siZNF695#1 and siZNF695#2 HCT-8 cells, and in Ctrl and ZNF695-overexpressing HT-29 cells. (F) Quantification colony numbers *P<0.05, **P<0.01vs. siCtrl, ## P<0.01 vs. Ctrl group. CRC, colorectal cancer; RT-qPCR, reverse transcription-quantitative PCR; CCK8, Cell Counting Kit-8; ZNF695, zinc finger protein 695; si, small interfering.

Article Snippet: The following primary antibodies were used: ZNF695 (cat. no. 25556-1-AP; 1:800; Proteintech Group, Inc.), NEK2 (cat. no. 24171-1-AP; 1:800; Proteintech Group Inc.), Akt (cat. no. 9272; 1:1,000; Cell Signaling Technology, Inc.), phosphorylated (p)-Akt (cat. no. 4060; 1:1,000; Cell Signaling Technology, Inc.), ribosomal protein S6 (S6; cat. no. 2217; 1:1,000; Cell Signaling Technology, Inc.), p-S6 (cat. no. 2211; 1:1,000; Cell Signaling Technology, Inc.), p21 (cat. no. 2947; 1:1,000; Cell Signaling Technology, Inc.), Bcl-2 (cat. no. 3498; 1:1,000; Cell Signaling Technology, Inc.), β-actin (cat. no. 66009-1-Ig; 1:5,000; Proteintech Group Inc.) and GAPDH (cat. no. 60004-1-Ig; 1:5,000; Proteintech Group Inc.).

Techniques: Over Expression, Quantitative RT-PCR, Western Blot, Transfection, CCK-8 Assay, Reverse Transcription, Real-time Polymerase Chain Reaction, Cell Counting

Journal: Oncology Reports

Article Title: Zinc finger protein 695 facilitates the proliferation of colorectal cancer cells through activation of the NEK2 and PI3K/Akt/mTOR signaling pathways

doi: 10.3892/or.2025.8949

Figure Lengend Snippet: ZNF695 modulates the apoptosis and cell cycle progression of CRC cells. (A) Cell cycle progression of siCtrl, siZNF695#1 and siZNF695#2 HCT-8 cells was measured by staining the cells with PI and via flow cytometric analysis. (B) Cell cycle progression of Ctrl and ZNF695-overexpressing HT-29 cells was detected by staining the cells with PI and via flow cytometric analysis. (C) Quantification of cell cycle results as shown in A and B. *P<0.05 vs. siCtrl, # P<0.05 vs. Ctrl group. (D) Apoptosis of siCtrl, siZNF695#1 and siZNF695#2 HCT-8 cells was assessed by staining the cells with Annexin V and PI, followed by flow cytometric analysis. (E) Apoptosis of Ctrl and ZNF695-overexpressing HT-29 cells was assessed by staining the cells with Annexin V and PI, followed by flow cytometric analysis. (F) Quantification of apoptosis results as shown in D and E. *P<0.05 vs. siCtrl, # P<0.05 vs. Ctrl group. (G) Caspase 3/7 activity was measured in HCT-8 cells transfected with siCtrl, siZNF695#1 and siZNF695#2 and in HT-29 cells infected with Ctrl and ZNF695-overexpressing lentiviruses. *P<0.05 vs. siCtrl, # P<0.05 vs. Ctrl group. CRC, colorectal cancer; ZNF695, zinc finger protein 695; si, small interfering.

Article Snippet: The following primary antibodies were used: ZNF695 (cat. no. 25556-1-AP; 1:800; Proteintech Group, Inc.), NEK2 (cat. no. 24171-1-AP; 1:800; Proteintech Group Inc.), Akt (cat. no. 9272; 1:1,000; Cell Signaling Technology, Inc.), phosphorylated (p)-Akt (cat. no. 4060; 1:1,000; Cell Signaling Technology, Inc.), ribosomal protein S6 (S6; cat. no. 2217; 1:1,000; Cell Signaling Technology, Inc.), p-S6 (cat. no. 2211; 1:1,000; Cell Signaling Technology, Inc.), p21 (cat. no. 2947; 1:1,000; Cell Signaling Technology, Inc.), Bcl-2 (cat. no. 3498; 1:1,000; Cell Signaling Technology, Inc.), β-actin (cat. no. 66009-1-Ig; 1:5,000; Proteintech Group Inc.) and GAPDH (cat. no. 60004-1-Ig; 1:5,000; Proteintech Group Inc.).

Techniques: Staining, Activity Assay, Transfection, Infection

Journal: Oncology Reports

Article Title: Zinc finger protein 695 facilitates the proliferation of colorectal cancer cells through activation of the NEK2 and PI3K/Akt/mTOR signaling pathways

doi: 10.3892/or.2025.8949

Figure Lengend Snippet: ZNF695 affects the expression of p21 and Bcl-2. (A) Western blot analysis of p21, Bcl-2 and GAPDH in (A) HCT-8 cells transfected with siCtrl, siZNF695#1 and siZNF695#2, and in (B) HT-29 cells infected with Ctrl and ZNF695-overexpressing lentiviruses. GAPDH served as internal control. ZNF695, zinc finger protein 695; si, small interfering.

Article Snippet: The following primary antibodies were used: ZNF695 (cat. no. 25556-1-AP; 1:800; Proteintech Group, Inc.), NEK2 (cat. no. 24171-1-AP; 1:800; Proteintech Group Inc.), Akt (cat. no. 9272; 1:1,000; Cell Signaling Technology, Inc.), phosphorylated (p)-Akt (cat. no. 4060; 1:1,000; Cell Signaling Technology, Inc.), ribosomal protein S6 (S6; cat. no. 2217; 1:1,000; Cell Signaling Technology, Inc.), p-S6 (cat. no. 2211; 1:1,000; Cell Signaling Technology, Inc.), p21 (cat. no. 2947; 1:1,000; Cell Signaling Technology, Inc.), Bcl-2 (cat. no. 3498; 1:1,000; Cell Signaling Technology, Inc.), β-actin (cat. no. 66009-1-Ig; 1:5,000; Proteintech Group Inc.) and GAPDH (cat. no. 60004-1-Ig; 1:5,000; Proteintech Group Inc.).

Techniques: Expressing, Western Blot, Transfection, Infection, Control

Journal: Oncology Reports

Article Title: Zinc finger protein 695 facilitates the proliferation of colorectal cancer cells through activation of the NEK2 and PI3K/Akt/mTOR signaling pathways

doi: 10.3892/or.2025.8949

Figure Lengend Snippet: ZNF695 positively regulates the expression of NEK2 in CRC cells. (A) List of genes that were positively correlated with ZNF695 in patients with CRC based on The Cancer Genome Atlas database. ***P<0.001. mRNA expression levels of NEK2 were analyzed by reverse transcription-qPCR in (B) HCT-8 cells transfected with siCtrl, siZNF695#1 and siZNF695#2, and in (C) HT-29 cells infected with Ctrl and ZNF695-overexpressing lentiviruses. *P<0.05 vs. siCtrl group; **P<0.01 vs. Ctrl group. Dual luciferase activity of the NEK2 gene promoter was detected in (D) HCT-8 cells transfected with siCtrl, siZNF695#1 and siZNF695#2, and in (E) HT-29 cells infected with Ctrl and ZNF695-overexpressing lentiviruses. *P<0.05 vs. siCtrl group; **P<0.01 vs. Ctrl group. (F) HT-29 cells were transfected with ZNF695-Flag-pCDNA3.1 vectors, and the cells were subjected to ChIP-qPCR experiments to assess whether ZNF695 interacted with the promoter sequence of the NEK2 gene. Based on The Cancer Genome Atlas database of patients with CRC. **P<0.01 vs. IgG group. (G) there was a positive correlation between ZNF695 and NEK2 (R=0.62; P<0.0001), and (H) a significant difference between the transcript levels of NEK2 in cancer (n=275) and non-cancer (n=349) tissues. *P<0.05. CRC, colorectal cancer; qPCR, quantitative PCR; ChIP, chromatin immunoprecipitation; ZNF695, zinc finger protein 695; si, small interfering; NEK2, NIMA-related kinase 2.

Article Snippet: The following primary antibodies were used: ZNF695 (cat. no. 25556-1-AP; 1:800; Proteintech Group, Inc.), NEK2 (cat. no. 24171-1-AP; 1:800; Proteintech Group Inc.), Akt (cat. no. 9272; 1:1,000; Cell Signaling Technology, Inc.), phosphorylated (p)-Akt (cat. no. 4060; 1:1,000; Cell Signaling Technology, Inc.), ribosomal protein S6 (S6; cat. no. 2217; 1:1,000; Cell Signaling Technology, Inc.), p-S6 (cat. no. 2211; 1:1,000; Cell Signaling Technology, Inc.), p21 (cat. no. 2947; 1:1,000; Cell Signaling Technology, Inc.), Bcl-2 (cat. no. 3498; 1:1,000; Cell Signaling Technology, Inc.), β-actin (cat. no. 66009-1-Ig; 1:5,000; Proteintech Group Inc.) and GAPDH (cat. no. 60004-1-Ig; 1:5,000; Proteintech Group Inc.).

Techniques: Expressing, Reverse Transcription, Transfection, Infection, Luciferase, Activity Assay, ChIP-qPCR, Sequencing, Real-time Polymerase Chain Reaction, Chromatin Immunoprecipitation

Journal: Oncology Reports

Article Title: Zinc finger protein 695 facilitates the proliferation of colorectal cancer cells through activation of the NEK2 and PI3K/Akt/mTOR signaling pathways

doi: 10.3892/or.2025.8949

Figure Lengend Snippet: Overexpression of ZNF695 activates the Akt/mTOR signaling pathway and dictates the sensitivity of CRC cells to rapamycin. (A) Western blot analysis of NEK2, p-Akt, Akt, p-S6, S6 and GAPDH in HCT-8 cells transfected with siCtrl, siZNF695#1 and siZNF695#2. GAPDH served as an internal control. (B) HCT-8 cells transfected with siCtrl, siZNF695#1 and siZNF695#2 were treated with different concentrations of rapamycin for 48 h and cell viability was determined by CCK8 assay. *P<0.05, **P<0.01 vs. siCtrl group. (C) Growth inhibition rate in response to different concentrations of rapamycin was calculated in HCT-8 cells transfected with siCtrl, siZNF695#1 and siZNF695#2. *P<0.05 vs. siCtrl group. (D) Western blot analysis of NEK2, p-Akt, Akt, p-S6, S6 and GAPDH in HT-29 cells infected with Ctrl and ZNF695-overexpressing lentiviruses. GAPDH served as an internal control. (E) HT-29 cells infected with Ctrl and ZNF695-overexpressing lentiviruses were treated with different concentrations of rapamycin for 48 h and cell viability was determined by CCK8 assay. *P<0.05, **P<0.01 vs. Ctrl group. (F) Growth inhibition rate in response to different concentrations of rapamycin was calculated in HT-29 cells infected with Ctrl and ZNF695-overexpressing lentiviruses. *P<0.05 vs. Ctrl group. CRC, colorectal cancer; CCK8, Cell Counting Kit-8; ZNF695, zinc finger protein 695; si, small interfering; p-, phosphorylated; NEK2, NIMA-related kinase 2; S6, ribosomal protein S6.

Article Snippet: The following primary antibodies were used: ZNF695 (cat. no. 25556-1-AP; 1:800; Proteintech Group, Inc.), NEK2 (cat. no. 24171-1-AP; 1:800; Proteintech Group Inc.), Akt (cat. no. 9272; 1:1,000; Cell Signaling Technology, Inc.), phosphorylated (p)-Akt (cat. no. 4060; 1:1,000; Cell Signaling Technology, Inc.), ribosomal protein S6 (S6; cat. no. 2217; 1:1,000; Cell Signaling Technology, Inc.), p-S6 (cat. no. 2211; 1:1,000; Cell Signaling Technology, Inc.), p21 (cat. no. 2947; 1:1,000; Cell Signaling Technology, Inc.), Bcl-2 (cat. no. 3498; 1:1,000; Cell Signaling Technology, Inc.), β-actin (cat. no. 66009-1-Ig; 1:5,000; Proteintech Group Inc.) and GAPDH (cat. no. 60004-1-Ig; 1:5,000; Proteintech Group Inc.).

Techniques: Over Expression, Western Blot, Transfection, Control, CCK-8 Assay, Inhibition, Infection, Cell Counting

Journal: Oncology Reports

Article Title: Zinc finger protein 695 facilitates the proliferation of colorectal cancer cells through activation of the NEK2 and PI3K/Akt/mTOR signaling pathways

doi: 10.3892/or.2025.8949

Figure Lengend Snippet: Knockdown of NEK2 rescues the tumor-promoting function of ZNF695. (A) Protein levels of NEK2 in Ctrl, ZNF695 and ZNF695 + siNEK2 HT-29 cells were detected by western blotting. (B) Proliferation of Ctrl, ZNF695 and ZNF695 + siNEK2 HT-29 cells was determined by Cell Counting Kit-8 assay. *P<0.05, **P<0.01 vs. Ctrl, # P<0.05, ## P<0.01 vs. ZNF695. (C) Colony formation was assess in Ctrl, ZNF695 and ZNF695 + siNEK2 HT-29 cells. *P<0.05 (Ctrl vs. ZNF695), # P<0.05 (ZNF695 vs. ZNF695 + siNEK2). (D) Caspase 3/7 activity was measured in Ctrl, ZNF695 and ZNF695 + siNEK2 HT-29 cells. *P<0.05, (Ctrl vs. ZNF695), # P<0.05 (ZNF695 vs. ZNF695 + siNEK2). CRC, colorectal cancer; ZNF695, zinc finger protein 695; si, small interfering; NEK2, NIMA-related kinase 2.

Article Snippet: The following primary antibodies were used: ZNF695 (cat. no. 25556-1-AP; 1:800; Proteintech Group, Inc.), NEK2 (cat. no. 24171-1-AP; 1:800; Proteintech Group Inc.), Akt (cat. no. 9272; 1:1,000; Cell Signaling Technology, Inc.), phosphorylated (p)-Akt (cat. no. 4060; 1:1,000; Cell Signaling Technology, Inc.), ribosomal protein S6 (S6; cat. no. 2217; 1:1,000; Cell Signaling Technology, Inc.), p-S6 (cat. no. 2211; 1:1,000; Cell Signaling Technology, Inc.), p21 (cat. no. 2947; 1:1,000; Cell Signaling Technology, Inc.), Bcl-2 (cat. no. 3498; 1:1,000; Cell Signaling Technology, Inc.), β-actin (cat. no. 66009-1-Ig; 1:5,000; Proteintech Group Inc.) and GAPDH (cat. no. 60004-1-Ig; 1:5,000; Proteintech Group Inc.).

Techniques: Knockdown, Western Blot, Cell Counting, Activity Assay

Journal: Oncology Reports

Article Title: Zinc finger protein 695 facilitates the proliferation of colorectal cancer cells through activation of the NEK2 and PI3K/Akt/mTOR signaling pathways

doi: 10.3892/or.2025.8949

Figure Lengend Snippet: Diagram of the ZNF695 signaling pathway in colorectal cancer tumorigenesis. ZNF695, zinc finger protein 695; NEK2, NIMA-related kinase 2.

Article Snippet: The following primary antibodies were used: ZNF695 (cat. no. 25556-1-AP; 1:800; Proteintech Group, Inc.), NEK2 (cat. no. 24171-1-AP; 1:800; Proteintech Group Inc.), Akt (cat. no. 9272; 1:1,000; Cell Signaling Technology, Inc.), phosphorylated (p)-Akt (cat. no. 4060; 1:1,000; Cell Signaling Technology, Inc.), ribosomal protein S6 (S6; cat. no. 2217; 1:1,000; Cell Signaling Technology, Inc.), p-S6 (cat. no. 2211; 1:1,000; Cell Signaling Technology, Inc.), p21 (cat. no. 2947; 1:1,000; Cell Signaling Technology, Inc.), Bcl-2 (cat. no. 3498; 1:1,000; Cell Signaling Technology, Inc.), β-actin (cat. no. 66009-1-Ig; 1:5,000; Proteintech Group Inc.) and GAPDH (cat. no. 60004-1-Ig; 1:5,000; Proteintech Group Inc.).

Techniques: