Journal: Oxidative Medicine and Cellular Longevity

Article Title: Depleted Long Noncoding RNA GAS5 Relieves Intervertebral Disc Degeneration via microRNA-17-3p/Ang-2

doi: 10.1155/2022/1792412

Figure Lengend Snippet: Silencing of GAS5 ameliorates IVDD in vivo by miR-17-3p-mediated inhibition of Ang-2. (a) Observation of the MRI signal intensity and IVDD degree by MRI scanning. (b) Western blot analysis of Ang-2 and ECM metabolism-related proteins in NP tissues extracted from IVDD mice, which was normalized to GAPDH. (c) Statistical results of TUNEL-positive cells in the NP tissues extracted from IVDD mice. (d) The expression of glucosaminoglycan (GAG) in mouse NP tissues detected by Safranin O-Fast Green staining. n = 8. (e) HE histological scores of mouse NP tissues by HE staining. n = 8. ∗ p < 0.05 vs. mice without any injection; # p < 0.05 vs. IVDD mice injected with harboring shRNA against shRNA NC; $ p < 0.05 vs. IVDD mice injected with lentivirus harboring shRNA against GAS5; $ p < 0.05 vs. IVDD mice injected with lentivirus harboring overexpression vector NC; @ p < 0.05 vs. IVDD mice injected with lentivirus harboring oe-GAS5. Data (mean ± standard deviation) were assessed by one-way ANOVA, followed by Tukey's post hoc test. n = 8.

Article Snippet: Afterwards, plasmids of sh-NC, sh-GAS5, oe-NC, oe-GAS5, miR-17-3p mimic, and miR-17-3p inhibitor were packaged into lentivirus vectors (pSIH1-H1-copGFP and pLV-EGFP-N) by GenePharma Co., Ltd. (Shanghai, China) to generate lentiviruses including pSIH1-H1-copGFP-sh-NC, pSIH1-H1-copGFP-sh-GAS5, pLV-EGFP-NC, pLV-EGFP-GAS5, pSIH1-H1-copGFP-miR-17-3p inhibitor, and pLV-EGFP-miR-17-3p, respectively.

Techniques: In Vivo, Inhibition, Western Blot, TUNEL Assay, Expressing, Staining, Injection, shRNA, Over Expression, Plasmid Preparation, Standard Deviation

Journal: American Journal of Cancer Research

Article Title: KLK11 suppresses cellular proliferation via inhibition of Wnt/β-catenin signaling pathway in esophageal squamous cell carcinoma

doi:

Figure Lengend Snippet: The expression of KLK11 is significantly lower in ESCC tissues. A and B. Indicated detection of KLK11 expression in dataset TCGA. C and D. Indicated KLK11 protein expression in 8 pairs of ESCC tissues and their para-carcinoma tissues by western blotting. KLK11 expression in ESCC tissues were significantly lower than those in the matched adjacent non-tumor tissues. E. KLK11 protein level was measured by immunohistochemical analysis in normal tissues. F. KLK11 protein level was measured by immunohistochemical analysis in ESCC tissues. Representative images are showed at 100 × and 200 × magnifications, respectively. G. KLK11 protein level in 100 ESCC tissues and matched normal tissues. Data were representative of three independent experiments (mean and SEM). *P < 0.05.

Article Snippet: Lentivirus vectors and transfection Three pairs of short hairpin RNA (shRNA) targeting KLK11 lentivirus vectors (shKLK11#1) and negative controls (scramble, NC) were specifically synthesized by Synbio Technologies (Suzhou, China).

Techniques: Expressing, Western Blot, Immunohistochemical staining

Journal: American Journal of Cancer Research

Article Title: KLK11 suppresses cellular proliferation via inhibition of Wnt/β-catenin signaling pathway in esophageal squamous cell carcinoma

doi:

Figure Lengend Snippet: KLK11 expression was significantly associated with clinical characteristics

Article Snippet: Lentivirus vectors and transfection Three pairs of short hairpin RNA (shRNA) targeting KLK11 lentivirus vectors (shKLK11#1) and negative controls (scramble, NC) were specifically synthesized by Synbio Technologies (Suzhou, China).

Techniques: Expressing

Journal: American Journal of Cancer Research

Article Title: KLK11 suppresses cellular proliferation via inhibition of Wnt/β-catenin signaling pathway in esophageal squamous cell carcinoma

doi:

Figure Lengend Snippet: Summary of univariate and multivariate Cox regression analyses of overall survival

Article Snippet: Lentivirus vectors and transfection Three pairs of short hairpin RNA (shRNA) targeting KLK11 lentivirus vectors (shKLK11#1) and negative controls (scramble, NC) were specifically synthesized by Synbio Technologies (Suzhou, China).

Techniques: Expressing

Journal: American Journal of Cancer Research

Article Title: KLK11 suppresses cellular proliferation via inhibition of Wnt/β-catenin signaling pathway in esophageal squamous cell carcinoma

doi:

Figure Lengend Snippet: Low KLK11 expression was associated with poor prognosis in ESCC patients. ESCC samples from the tissue microarray were divided as two different groups, high KLK11 expression and low KLK11 expression group. Patients’ survival information was provided by the manufacture. Kaplan-Meier analysis of overall survival was done for the two group.

Article Snippet: Lentivirus vectors and transfection Three pairs of short hairpin RNA (shRNA) targeting KLK11 lentivirus vectors (shKLK11#1) and negative controls (scramble, NC) were specifically synthesized by Synbio Technologies (Suzhou, China).

Techniques: Expressing, Microarray

Journal: American Journal of Cancer Research

Article Title: KLK11 suppresses cellular proliferation via inhibition of Wnt/β-catenin signaling pathway in esophageal squamous cell carcinoma

doi:

Figure Lengend Snippet: The expression of KLK11 is significantly lower in ESCC cell lines. QPCR (A) and western blotting (B) showed the expression of KLK11 in seven ESCC cell lines and HEEC. KLK11 knockdown efficiency was confirmed by qPCR (C) and western blotting (D) in ESCC cells. qPCR (E) and western blotting (F) confirmed the restoration of KLK11 expression in four groups of TE-1 cells. qPCR (G) and western blotting (H) confirmed the restoration of KLK11 expression in four groups of EC18 cells. Results were showed as mean ± SD (*P < 0.05) of triplicate determination from three independent experiments.

Article Snippet: Lentivirus vectors and transfection Three pairs of short hairpin RNA (shRNA) targeting KLK11 lentivirus vectors (shKLK11#1) and negative controls (scramble, NC) were specifically synthesized by Synbio Technologies (Suzhou, China).

Techniques: Expressing, Western Blot, Knockdown

Journal: American Journal of Cancer Research

Article Title: KLK11 suppresses cellular proliferation via inhibition of Wnt/β-catenin signaling pathway in esophageal squamous cell carcinoma

doi:

Figure Lengend Snippet: KLK11 inhibited the proliferation of ESCC cells. A. KLK11 inhibited the proliferation of TE-1 cells by MTT assays. B. KLK11 inhibited the proliferation of EC18 cells by MTT assays. C. Colony formation assays showed that KLK11 overexpression inhibited TE-1 and EC18 cells proliferation. These effects were abolished by treatment with shKLK11. D. Flow cytometry analysis of cell cycle phase revealed a G0/G1 arrest induced by KLK11. Data were representative of three independent experiments (mean and SEM). *P < 0.05.

Article Snippet: Lentivirus vectors and transfection Three pairs of short hairpin RNA (shRNA) targeting KLK11 lentivirus vectors (shKLK11#1) and negative controls (scramble, NC) were specifically synthesized by Synbio Technologies (Suzhou, China).

Techniques: Over Expression, Flow Cytometry

Journal: American Journal of Cancer Research

Article Title: KLK11 suppresses cellular proliferation via inhibition of Wnt/β-catenin signaling pathway in esophageal squamous cell carcinoma

doi:

Figure Lengend Snippet: KLK11 affected the expression of the related factors in Wnt/β-catenin pathway and cell cycle-mediated factors. A. The expression of p-GSK-3β/GSK-3β in EC18 and TE-1 cells after lentivirus vectors delivery in each group as measured by western blot. B. The TCF/LEF luciferase ratio reported the activity of Wnt/β-catenin pathway in the indicated cells. C. Expression of CDK4, cyclin D1, CDK6, p-Rb, Rb, β-catenin (cytosol), and β-catenin (nucleus) were determined in TE-1 and EC18 cells transfected with KLK11. KLK11 inhibited the proliferation of ESCC by regulating the expression of the above proteins. D. Analysis of Ki67, cyclin D1, CDK4, CDK6, and c-myc expression in KLK11-overexpression or KLK11-knockdown ESCC cells by qPCR. Data were representative of three independent experiments (mean and SEM). *P < 0.05.

Article Snippet: Lentivirus vectors and transfection Three pairs of short hairpin RNA (shRNA) targeting KLK11 lentivirus vectors (shKLK11#1) and negative controls (scramble, NC) were specifically synthesized by Synbio Technologies (Suzhou, China).

Techniques: Expressing, Western Blot, Luciferase, Activity Assay, Transfection, Over Expression, Knockdown

Journal: American Journal of Cancer Research

Article Title: KLK11 suppresses cellular proliferation via inhibition of Wnt/β-catenin signaling pathway in esophageal squamous cell carcinoma

doi:

Figure Lengend Snippet: KLK11 down regulated the expression of Ki67 and cyclin D1 through deactivation of the Wnt/β-catenin signaling pathway. A, B. ESCC cells were pre-treated with XAV-939 for 24 h before harvesting, and the detection of Ki67, cyclin D1, c-myc, and expression levels were performed by qPCR. C, D. Treatment of EC18 and TE-1 cells with XAV-939, the expression of Ki67, cyclin D1, β-catenin (cytosol), and β-catenin (nucleus) were significantly inhibited. Data were representative of three independent experiments (mean and SEM). *P < 0.05.

Article Snippet: Lentivirus vectors and transfection Three pairs of short hairpin RNA (shRNA) targeting KLK11 lentivirus vectors (shKLK11#1) and negative controls (scramble, NC) were specifically synthesized by Synbio Technologies (Suzhou, China).

Techniques: Expressing

Journal: American Journal of Cancer Research

Article Title: KLK11 suppresses cellular proliferation via inhibition of Wnt/β-catenin signaling pathway in esophageal squamous cell carcinoma

doi:

Figure Lengend Snippet: KLK11 suppressed ESCC cells proliferation through inhibiting the activity of the Wnt/β-catenin pathway. A, B. XAV-939 was added to TE-1 and EC18 cells transfected with shRNA, and the proliferation of TE-1 and EC18 cells was detected by MTT assays. C. XAV-939 was added to TE-1 and EC18 cells transfected with shRNA, and the proliferation of TE-1 and EC18 cells was detected by colony formation assays. D. The relationship between KLK11 and the Wnt/β-catenin pathway. Data were representative of three independent experiments (mean and SEM). *P < 0.05.

Article Snippet: Lentivirus vectors and transfection Three pairs of short hairpin RNA (shRNA) targeting KLK11 lentivirus vectors (shKLK11#1) and negative controls (scramble, NC) were specifically synthesized by Synbio Technologies (Suzhou, China).

Techniques: Activity Assay, Transfection, shRNA

Journal: American Journal of Cancer Research

Article Title: KLK11 suppresses cellular proliferation via inhibition of Wnt/β-catenin signaling pathway in esophageal squamous cell carcinoma

doi:

Figure Lengend Snippet: KLK11 inhibited the proliferation of ESCC in vivo. (A, B) Tumor xenograft volume (A) and tumor size (B) in shRNA-KLK11-treated nude mice were bigger than those in the scramble group. Tumor sizes in KLK11 overexpression nude mice were smaller than those in the vector group. (C) Analysis of Ki67, cyclin D1, and KLK11 expression in mice tumor tissues with KLK11 overexpression or KLK11 knockdown by qPCR. (D) Analysis of KLK11 and Cyclin D1 in xenografts by Immunohistochemistry. Data were representative of three independent experiments (mean and SEM). *P < 0.05.

Article Snippet: Lentivirus vectors and transfection Three pairs of short hairpin RNA (shRNA) targeting KLK11 lentivirus vectors (shKLK11#1) and negative controls (scramble, NC) were specifically synthesized by Synbio Technologies (Suzhou, China).

Techniques: In Vivo, shRNA, Over Expression, Plasmid Preparation, Expressing, Knockdown, Immunohistochemistry