Journal: Cancer Management and Research

Article Title: KLF16 Downregulates the Expression of Tumor Suppressor Gene TGFBR3 to Promote Bladder Cancer Proliferation and Migration

doi: 10.2147/CMAR.S334521

Figure Lengend Snippet: KLF16 is highly expressed in BC specimens. ( A ) The levels of KLF16 transcript were evaluated in 404 of bladder urothelial carcinoma (BLCA) and 28 normal samples from the TCGA database, p<0.05. ( B ) Immunohistochemical staining of KLF16 in 12 pairs of BC and normal samples. The quantitative result is presented. ***p<0.001. ( C ) qRT-PCR detection of KLF16 in SV-HUC-1, T24, 5637, and RT-4 cells. The expression of KLF16 was adjusted to β-actin expression. *p<0.05. **p<0.01. ( D ) BLCA patients were grouped into high- and low-expression KLF16 groups according to TCGA database. Overall survival was analyzed in the patients. p=0.013.

Article Snippet: The human normal bladder epithelial cell line SV-HUC-1 and the BC cell lines T24, 5637, and RT-4 were obtained from ATCC.

Techniques: Immunohistochemical staining, Staining, Quantitative RT-PCR, Expressing

Journal: Cancer Management and Research

Article Title: KLF16 Downregulates the Expression of Tumor Suppressor Gene TGFBR3 to Promote Bladder Cancer Proliferation and Migration

doi: 10.2147/CMAR.S334521

Figure Lengend Snippet: KLF16 is essential for BC cell growth and migration. ( A ) Immunoblotting analysis of KLF16 in siCtrl, siKLF16#1 and siKLF16#2-transfected 5637 and T24 cells. Expression of KLF16 was adjusted to GAPDH expression. ( B ) Cell proliferation was examined in siCtrl, siKLF16#1, and siKLF16#2-treated 5637 and T24 cells. siCtrl vs siKLF16#1 **p<0.01 and siCtrl vs siKLF16#1 ***p<0.001, siCtrl vs siKLF16#2 ##p<0.01, siCtrl vs siKLF16#2 ###p<0.001. ( C ) Colony growth was evaluated in siCtrl, siKLF16#1, and siKLF16#2-treated 5637 and T24 cells. **p<0.01, ***p<0.001. ( D and E ) Cell apoptosis was detected by PI/Annexin V staining in siCtrl, siKLF16#1 and siKLF16#2-treated 5637 and T24 cells, respectively. ***p<0.001. ( F ) Cell migration was assessed using Transwell assays using 5637 and T24 cells exposed to siCtrl, siKLF16#1, and siKLF16#2, respectively. **p<0.01, ***p<0.001.

Article Snippet: The human normal bladder epithelial cell line SV-HUC-1 and the BC cell lines T24, 5637, and RT-4 were obtained from ATCC.

Techniques: Migration, Western Blot, Transfection, Expressing, Staining

Journal: Cancer Management and Research

Article Title: KLF16 Downregulates the Expression of Tumor Suppressor Gene TGFBR3 to Promote Bladder Cancer Proliferation and Migration

doi: 10.2147/CMAR.S334521

Figure Lengend Snippet: Overexpression of KLF16 promoted BC cell growth and migration. ( A ) Immunoblot analysis of KLF16 in 5637 and T24 cells overexpressing Ctrl and KLF16. Expression of KLF16 was adjusted to GAPDH expression. ( B ) Cell proliferation was examined in 5637 and T24 cells overexpressing Ctrl and KLF16. **p<0.01, ***p<0.001. ( C ) Colony growth was evaluated in 5637 and T24 cells overexpressing Ctrl and KLF16. **p<0.01, ***p<0.001. ( D and E ) Cell apoptosis was detected by PI/Annexin V staining in Ctrl and KLF16 overexpressing 5637 and T24 cells. ***p<0.001. ( F ) Cell migration was detected by the Transwell assay in 5637 and T24 cells overexpressing Ctrl and KLF16. **p<0.01, ***p<0.001.

Article Snippet: The human normal bladder epithelial cell line SV-HUC-1 and the BC cell lines T24, 5637, and RT-4 were obtained from ATCC.

Techniques: Over Expression, Migration, Western Blot, Expressing, Staining, Transwell Assay

Journal: Cancer Management and Research

Article Title: KLF16 Downregulates the Expression of Tumor Suppressor Gene TGFBR3 to Promote Bladder Cancer Proliferation and Migration

doi: 10.2147/CMAR.S334521

Figure Lengend Snippet: KLF16 negatively regulates TGFBR3 by inhibiting its transcriptional activity. ( A and B ) mRNA and protein were determined by qRT-PCR and immunoblotting and normalized to β-actin in KLF16 silencing and overexpressing cells. ( C ) 293T cells were transfected with pCDNA3.1-Ctrl or pCDNA3.1-KLF16, pGL3. Basic-TGFBR3 promoter, and TK vectors. 48 hours later, dual luciferase activity was assessed and adjusted for TK. **p<0.01. ( D ) The ChIP-qPCR assay was performed in T24 and 5637 cells overexpressing KLF16-flag. Relative enrichment was adjusted to the IgG group. **p<0.01. ( E ) TGFBR3 transcript levels in 404 BC and 28 normal samples were assessed using datasets obtained from TCGA database. *p<0.05. ( F ) Spearman correlation between KLF16 and TGFBR3 was analyzed in BLCA samples from TCGA database. p<0.0001.

Article Snippet: The human normal bladder epithelial cell line SV-HUC-1 and the BC cell lines T24, 5637, and RT-4 were obtained from ATCC.

Techniques: Activity Assay, Quantitative RT-PCR, Western Blot, Transfection, Luciferase, ChIP-qPCR

Journal: Cancer Management and Research

Article Title: KLF16 Downregulates the Expression of Tumor Suppressor Gene TGFBR3 to Promote Bladder Cancer Proliferation and Migration

doi: 10.2147/CMAR.S334521

Figure Lengend Snippet: Downregulation of TGFBR3 by KLF16 contributes to BC growth and migration. ( A – D ) T24 cells were transfected with lentivirus overexpressing Ctrl, KLF16, and KLF16+TGFBR3. The cells were subjected to immunoblotting analysis of TGFBR3 ( A ), CCK8 assay ( B ), Transwell assay ( C ), and PI/Annexin V-FITC staining of apoptosis ( D ). Quantification results were presented. **p<0.01. ( E – H ) 5637 cells were transfected with siRNAs against Ctrl, KLF16 and KLF16+TGFBR3. The cells were subjected to immunoblotting analysis of TGFBR3 ( E ), CCK8 assay ( F ), colony formation ( G ), and Transwell assay ( H ). Quantification results were presented. **p<0.01.

Article Snippet: The human normal bladder epithelial cell line SV-HUC-1 and the BC cell lines T24, 5637, and RT-4 were obtained from ATCC.

Techniques: Migration, Transfection, Western Blot, CCK-8 Assay, Transwell Assay, Staining

Journal: Cancer Management and Research

Article Title: KLF16 Downregulates the Expression of Tumor Suppressor Gene TGFBR3 to Promote Bladder Cancer Proliferation and Migration

doi: 10.2147/CMAR.S334521

Figure Lengend Snippet: Downregulation of TGFBR3 by KLF16 promotes BC tumorigenesis. ( A ) Macroscopic images of xenografted tumors of T24 cells transfected with the lentivirus Ctrl (negative control), KLF16 (KLF16 overexpression), and KLF16+TGFBR3 (KLF16 and TGFBR3 overexpression) lentivirus. ( B ) Tumor growth was evaluated on day 14 after implantation. ns, not significant. **p<0.01. ( C ) Tumors were weighed on day 36 after implantation. **p<0.01. ( D ) qRT-PCR analysis of KLF16, TGFBR3, and PCNA in tumors. *p<0.05.

Article Snippet: The human normal bladder epithelial cell line SV-HUC-1 and the BC cell lines T24, 5637, and RT-4 were obtained from ATCC.

Techniques: Transfection, Negative Control, Over Expression, Quantitative RT-PCR