Journal: Frontiers in Cell and Developmental Biology

Article Title: CORO6 Promotes Cell Growth and Invasion of Clear Cell Renal Cell Carcinoma via Activation of WNT Signaling

doi: 10.3389/fcell.2021.647301

Figure Lengend Snippet: Experimental validation of CORO6 expression in ccRCC samples and RCC cell lines. (A) IHC staining showed that CORO6 was overexpressed in ccRCC tumors. Top, representative images of CORO6 IHC staining in tissue microarrays. Bottom, quantification of CORO6 IHC staining in tissue microarrays with ccRCC tumors ( n = 75) and adjacent normal kidney tissues ( n = 75). (B) The CORO6 mRNA level was considerably increased in our collected ccRCC samples ( n = 40) compared to paired normal tissues ( n = 40). 18s rRNA served as the normalizing control. (C) The CORO6 protein level was significantly increased in our collected ccRCC samples ( n = 40) compared to paired normal tissues ( n = 40). (C) Western blotting showed that ccRCC tumors expressed a high level of CORO6. GAPDH served as the internal control. (D) The qPCR assay exhibited that RCC cell lines had a higher level of CORO6 compared to normal HK2 cells. Gene expression was normalized to 18s rRNA. (E) Western blotting confirmed that the CORO6 protein level was increased in RCC cell lines. GAPDH served as the loading control. (F) A quantification of (E) . * P < 0.05, ** P < 0.01, *** P < 0.001, and **** P < 0.0001. Ns, no significance.

Article Snippet: The antibodies used in this study were as follows: c-Myc (10828-1-AP; Proteintech, Rosemont, IL, USA), CORO6 (17243-1-AP; Proteintech), CCND1 (60186-1-lg; Proteintech), GAPDH (60004-lg; Proteintech), and AXIN2 (ab109307; Abcam, Cambridge, UK).

Techniques: Biomarker Discovery, Expressing, Immunohistochemistry, Control, Western Blot, Gene Expression

Journal: Frontiers in Cell and Developmental Biology

Article Title: CORO6 Promotes Cell Growth and Invasion of Clear Cell Renal Cell Carcinoma via Activation of WNT Signaling

doi: 10.3389/fcell.2021.647301

Figure Lengend Snippet: CORO6 enhanced ccRCC cell growth. (A,B) Knockdown efficiency of shRNAs against CORO6 by qPCR (A) and Western blotting (B) . (C) The MTT assay revealed that CORO6 reduction suppressed Caki-1 and SN12-PM6 cell growth. (D) EdU staining showed that CORO6 reduction decreased Caki-1 and SN12-PM6 cell proliferation. Top: representative images of EdU staining. Bottom: statistical analyses. (E) PI-stained flow cytometry analysis showed that CORO6 depletion caused cell cycle arrest at G0/G1 in Caki-1 and SN12-PM6 cells. (F) Annexin V-/PI-stained flow cytometry analysis indicated that CORO6 depletion increased Caki-1 and SN12-PM6 cell apoptosis. (G) Western blotting analyses revealed that CORO6 knockdown increased expression levels of cleaved PRAP and cleaved caspase-3. Top, representative image of western blotting analyses. Bottom, a statistical analyses. GAPDH was a loading control. * P < 0.05, ** P < 0.01, *** P < 0.001, and **** P < 0.0001.

Article Snippet: The antibodies used in this study were as follows: c-Myc (10828-1-AP; Proteintech, Rosemont, IL, USA), CORO6 (17243-1-AP; Proteintech), CCND1 (60186-1-lg; Proteintech), GAPDH (60004-lg; Proteintech), and AXIN2 (ab109307; Abcam, Cambridge, UK).

Techniques: Knockdown, Western Blot, MTT Assay, Staining, Flow Cytometry, Expressing, Control

Journal: Frontiers in Cell and Developmental Biology

Article Title: CORO6 Promotes Cell Growth and Invasion of Clear Cell Renal Cell Carcinoma via Activation of WNT Signaling

doi: 10.3389/fcell.2021.647301

Figure Lengend Snippet: CORO6-induced ccRCC cell growth and cell invasion/migration were dependent on WNT activation. (A) IWP-O1 had no effect on the CORO6 mRNA level. 18s rRNA was used as the normalizing control. (B) IWP-O1 had no effect on the CORO6 protein level. GAPDH served as the loading control. (C) The MTT assay showed that IWP-O1 blocked CORO6-induced cell growth of Caki-1 cells. (D) EdU staining revealed that IWP-O1 inhibited CORO6-increased cell proliferation of Caki-1 cells. Left: representative image of EdU staining. Right: statistical analysis. (E) PI-stained flow cytometry analysis showed that CORO6-mediated cell cycle progression from G0/G1 to the S phase of Caki-1 cells was blocked by IWP-O1. Left: representative image of PI-stained flow cytometry analysis. Right: statistical analysis. (F) Annexin V-/PI-stained flow cytometry analysis indicated that CORO6-decreased cell apoptosis of Caki-1 cells was reversed by IWP-O1 treatment. Left: representative image of annexin V-/PI-stained flow cytometry analysis. Right: statistical analysis. (G) CORO6-induced Caki-1 cell migration was suppressed by IWP-O1 treatment. Left: representative images of migrating cells. Right: statistical analysis of migrating cells. (H) CORO6-induced Caki-1 cell invasion was suppressed by IWP-O1 treatment. Left: representative images of invading cells. Right: statistical analysis. ** P < 0.01, *** P < 0.001, and **** P < 0.0001. Compared with CORO6 OE, # P < 0.05, ## P < 0.01, ### P < 0.001, and #### P < 0.0001.

Article Snippet: The antibodies used in this study were as follows: c-Myc (10828-1-AP; Proteintech, Rosemont, IL, USA), CORO6 (17243-1-AP; Proteintech), CCND1 (60186-1-lg; Proteintech), GAPDH (60004-lg; Proteintech), and AXIN2 (ab109307; Abcam, Cambridge, UK).

Techniques: Migration, Activation Assay, Control, MTT Assay, Staining, Flow Cytometry

Journal: Frontiers in Cell and Developmental Biology

Article Title: CORO6 Promotes Cell Growth and Invasion of Clear Cell Renal Cell Carcinoma via Activation of WNT Signaling

doi: 10.3389/fcell.2021.647301

Figure Lengend Snippet: Xenograft mouse model to support the oncogenic role of CORO6 in ccRCC development. (A–C) CORO6 depletion suppressed ccRCC tumor growth monitored by tumor size (A) , growth curve (B) , and tumor weight (C) . (D) The mRNA levels of c-Myc, AXIN2, and CCND1 were remarkably reduced in CORO6-depleted ccRCC tumors. The expression levels of detected genes were normalized to 18s rRNA. (E) Western blotting showed that the protein levels of c-Myc, AXIN2, and CCND1 were remarkably reduced in CORO6-depleted ccRCC tumors. GAPDH was used as the internal control. (F–H) CORO6-promoted ccRCC tumor growth was inhibited by the administration of IWP-O1 monitored by tumor size (F) , growth curve (G) , and tumor weight (H) . ** P < 0.01, *** P < 0.001, and **** P < 0.0001. Compared with CORO6 OE, ### p < 0.001 and #### p < 0.0001.

Article Snippet: The antibodies used in this study were as follows: c-Myc (10828-1-AP; Proteintech, Rosemont, IL, USA), CORO6 (17243-1-AP; Proteintech), CCND1 (60186-1-lg; Proteintech), GAPDH (60004-lg; Proteintech), and AXIN2 (ab109307; Abcam, Cambridge, UK).

Techniques: Expressing, Western Blot, Control

Journal: Frontiers in Cell and Developmental Biology

Article Title: CORO6 Promotes Cell Growth and Invasion of Clear Cell Renal Cell Carcinoma via Activation of WNT Signaling

doi: 10.3389/fcell.2021.647301

Figure Lengend Snippet: CORO6 upregulation in ccRCC patients. (A) The CORO6 mRNA level was significantly increased in ccRCC patients ( n = 533) compared to normal kidney tissues ( n = 72). (B) The CORO6 mRNA level was indistinguishable between ccRCC ( n = 72) and paired adjacent tissues ( n = 72). (C) The CORO6 expression level displayed no correlation with sex hormones. (D) Top: the CORO6 mRNA level was upregulated in ccRCC patients with T stage III+IV compared to patients with T stage I+II. Bottom: ccRCC patients with T stage III tended to express a higher level of CORO6 compared to patients with T stage I. (E) Top: the CORO6 mRNA level was elevated in ccRCC patients with TNM stage III+IV compared to patients with TNM stage I+II. Bottom: ccRCC patients with TNM stage III or IV tended to express a higher level of CORO6 when compared to patients with TNM stage I. (F) Top: the CORO6 mRNA level was upregulated in ccRCC patients with G3+G4 compared to patients with G1+G2. Bottom: ccRCC patients with G4 tended to express a higher level of CORO6 compared to G2+G3. One way ANOVA test was performed in bottom panels of (D–F) . * P < 0.05, ** P < 0.01, and *** P < 0.001. Ns, no significance.

Article Snippet: The slides were incubated with endogenous peroxidase blocking solution, and then incubated with the CORO6 (17243-1-AP, Proteintech) antibody at 4°C overnight.

Techniques: Expressing

Journal: Frontiers in Cell and Developmental Biology

Article Title: CORO6 Promotes Cell Growth and Invasion of Clear Cell Renal Cell Carcinoma via Activation of WNT Signaling

doi: 10.3389/fcell.2021.647301

Figure Lengend Snippet: High CORO6 level was an independent predictor of poor prognosis in ccRCC patients. A high expression level of CORO6 is strongly associated with tumor metastasis (A,B) , poor OS (C) , and death (D) . * P < 0.05 and *** P < 0.001. Ns, no significance.

Article Snippet: The slides were incubated with endogenous peroxidase blocking solution, and then incubated with the CORO6 (17243-1-AP, Proteintech) antibody at 4°C overnight.

Techniques: Expressing

Journal: Frontiers in Cell and Developmental Biology

Article Title: CORO6 Promotes Cell Growth and Invasion of Clear Cell Renal Cell Carcinoma via Activation of WNT Signaling

doi: 10.3389/fcell.2021.647301

Figure Lengend Snippet: CORO6 level was highly correlated with the OS of ccRCC patients. (A) CcRCC patients with a high expression level of CORO6 had a shorter OS time. (B–L) OS analyses of CORO6 in ccRCC patients classified by female (B) , male (C) , age > 65 (D) , age ≤ 65 (E) , T1+T2 (F) , T3+T4 (G) , N0 (H) , M0 (J) , M1 (K) , TNM stage III+IV (M) , and G3+G4 (L) .

Article Snippet: The slides were incubated with endogenous peroxidase blocking solution, and then incubated with the CORO6 (17243-1-AP, Proteintech) antibody at 4°C overnight.

Techniques: Expressing

Journal: Frontiers in Cell and Developmental Biology

Article Title: CORO6 Promotes Cell Growth and Invasion of Clear Cell Renal Cell Carcinoma via Activation of WNT Signaling

doi: 10.3389/fcell.2021.647301

Figure Lengend Snippet: Experimental validation of CORO6 expression in ccRCC samples and RCC cell lines. (A) IHC staining showed that CORO6 was overexpressed in ccRCC tumors. Top, representative images of CORO6 IHC staining in tissue microarrays. Bottom, quantification of CORO6 IHC staining in tissue microarrays with ccRCC tumors ( n = 75) and adjacent normal kidney tissues ( n = 75). (B) The CORO6 mRNA level was considerably increased in our collected ccRCC samples ( n = 40) compared to paired normal tissues ( n = 40). 18s rRNA served as the normalizing control. (C) The CORO6 protein level was significantly increased in our collected ccRCC samples ( n = 40) compared to paired normal tissues ( n = 40). (C) Western blotting showed that ccRCC tumors expressed a high level of CORO6. GAPDH served as the internal control. (D) The qPCR assay exhibited that RCC cell lines had a higher level of CORO6 compared to normal HK2 cells. Gene expression was normalized to 18s rRNA. (E) Western blotting confirmed that the CORO6 protein level was increased in RCC cell lines. GAPDH served as the loading control. (F) A quantification of (E) . * P < 0.05, ** P < 0.01, *** P < 0.001, and **** P < 0.0001. Ns, no significance.

Article Snippet: The slides were incubated with endogenous peroxidase blocking solution, and then incubated with the CORO6 (17243-1-AP, Proteintech) antibody at 4°C overnight.

Techniques: Biomarker Discovery, Expressing, Immunohistochemistry, Control, Western Blot, Gene Expression

Journal: Frontiers in Cell and Developmental Biology

Article Title: CORO6 Promotes Cell Growth and Invasion of Clear Cell Renal Cell Carcinoma via Activation of WNT Signaling

doi: 10.3389/fcell.2021.647301

Figure Lengend Snippet: Correlation between clinicopathological variables and CORO6 expression in RCC.

Article Snippet: The slides were incubated with endogenous peroxidase blocking solution, and then incubated with the CORO6 (17243-1-AP, Proteintech) antibody at 4°C overnight.

Techniques: Expressing

Journal: Frontiers in Cell and Developmental Biology

Article Title: CORO6 Promotes Cell Growth and Invasion of Clear Cell Renal Cell Carcinoma via Activation of WNT Signaling

doi: 10.3389/fcell.2021.647301

Figure Lengend Snippet: CORO6 enhanced ccRCC cell growth. (A,B) Knockdown efficiency of shRNAs against CORO6 by qPCR (A) and Western blotting (B) . (C) The MTT assay revealed that CORO6 reduction suppressed Caki-1 and SN12-PM6 cell growth. (D) EdU staining showed that CORO6 reduction decreased Caki-1 and SN12-PM6 cell proliferation. Top: representative images of EdU staining. Bottom: statistical analyses. (E) PI-stained flow cytometry analysis showed that CORO6 depletion caused cell cycle arrest at G0/G1 in Caki-1 and SN12-PM6 cells. (F) Annexin V-/PI-stained flow cytometry analysis indicated that CORO6 depletion increased Caki-1 and SN12-PM6 cell apoptosis. (G) Western blotting analyses revealed that CORO6 knockdown increased expression levels of cleaved PRAP and cleaved caspase-3. Top, representative image of western blotting analyses. Bottom, a statistical analyses. GAPDH was a loading control. * P < 0.05, ** P < 0.01, *** P < 0.001, and **** P < 0.0001.

Article Snippet: The slides were incubated with endogenous peroxidase blocking solution, and then incubated with the CORO6 (17243-1-AP, Proteintech) antibody at 4°C overnight.

Techniques: Knockdown, Western Blot, MTT Assay, Staining, Flow Cytometry, Expressing, Control

Journal: Frontiers in Cell and Developmental Biology

Article Title: CORO6 Promotes Cell Growth and Invasion of Clear Cell Renal Cell Carcinoma via Activation of WNT Signaling

doi: 10.3389/fcell.2021.647301

Figure Lengend Snippet: CORO6 promoted ccRCC cell migration and invasion. (A) The wound healing assay showed that knockdown of CORO6 inhibited Caki-1 and SN12-PM6 cell migration. (B,C) The transwell migration assay revealed that CORO6 knockdown suppressed Caki-1 and SN12-PM6 cell migration. (B) Representative images of migrating cells. (C) Statistical analysis of (B) . (D,E) CORO6 knockdown significantly suppressed the invasion ability of Caki-1 and SN12-PM6 cells. (D) Representative images of invading cells. (E) Statistical analysis of (D) . *** P < 0.001.

Article Snippet: The slides were incubated with endogenous peroxidase blocking solution, and then incubated with the CORO6 (17243-1-AP, Proteintech) antibody at 4°C overnight.

Techniques: Migration, Wound Healing Assay, Knockdown, Transwell Migration Assay

Journal: Frontiers in Cell and Developmental Biology

Article Title: CORO6 Promotes Cell Growth and Invasion of Clear Cell Renal Cell Carcinoma via Activation of WNT Signaling

doi: 10.3389/fcell.2021.647301

Figure Lengend Snippet: Mechanistic dissection of how CORO6 promoted ccRCC progression. (A) GSEA KEGG analyses showed that GO receptor agonist activity was highly enriched in the high CORO6 group. (B) The expression levels of WNT1, WNT3, and WNT10B increased in ccRCC patients with a high level of CORO6. (C,D) CORO6 knockdown dramatically reduced the expression levels of WNT1, WNT3, and WNT10B in both Caki-1 (C) and SN12-PM6 cells (D) . The expression levels of detected genes were normalized to 18s rRNA. (E,F) CORO6 enhanced the activity of TOPflash but not FOPflash in both Caki-1 (E) and SN12-PM6 cells (F) . * P < 0.05, *** P < 0.001, and **** P < 0.0001. Ns, no significance.

Article Snippet: The slides were incubated with endogenous peroxidase blocking solution, and then incubated with the CORO6 (17243-1-AP, Proteintech) antibody at 4°C overnight.

Techniques: Dissection, Activity Assay, Expressing, Knockdown

Journal: Frontiers in Cell and Developmental Biology

Article Title: CORO6 Promotes Cell Growth and Invasion of Clear Cell Renal Cell Carcinoma via Activation of WNT Signaling

doi: 10.3389/fcell.2021.647301

Figure Lengend Snippet: CORO6-induced ccRCC cell growth and cell invasion/migration were dependent on WNT activation. (A) IWP-O1 had no effect on the CORO6 mRNA level. 18s rRNA was used as the normalizing control. (B) IWP-O1 had no effect on the CORO6 protein level. GAPDH served as the loading control. (C) The MTT assay showed that IWP-O1 blocked CORO6-induced cell growth of Caki-1 cells. (D) EdU staining revealed that IWP-O1 inhibited CORO6-increased cell proliferation of Caki-1 cells. Left: representative image of EdU staining. Right: statistical analysis. (E) PI-stained flow cytometry analysis showed that CORO6-mediated cell cycle progression from G0/G1 to the S phase of Caki-1 cells was blocked by IWP-O1. Left: representative image of PI-stained flow cytometry analysis. Right: statistical analysis. (F) Annexin V-/PI-stained flow cytometry analysis indicated that CORO6-decreased cell apoptosis of Caki-1 cells was reversed by IWP-O1 treatment. Left: representative image of annexin V-/PI-stained flow cytometry analysis. Right: statistical analysis. (G) CORO6-induced Caki-1 cell migration was suppressed by IWP-O1 treatment. Left: representative images of migrating cells. Right: statistical analysis of migrating cells. (H) CORO6-induced Caki-1 cell invasion was suppressed by IWP-O1 treatment. Left: representative images of invading cells. Right: statistical analysis. ** P < 0.01, *** P < 0.001, and **** P < 0.0001. Compared with CORO6 OE, # P < 0.05, ## P < 0.01, ### P < 0.001, and #### P < 0.0001.

Article Snippet: The slides were incubated with endogenous peroxidase blocking solution, and then incubated with the CORO6 (17243-1-AP, Proteintech) antibody at 4°C overnight.

Techniques: Migration, Activation Assay, Control, MTT Assay, Staining, Flow Cytometry

Journal: Frontiers in Cell and Developmental Biology

Article Title: CORO6 Promotes Cell Growth and Invasion of Clear Cell Renal Cell Carcinoma via Activation of WNT Signaling

doi: 10.3389/fcell.2021.647301

Figure Lengend Snippet: Xenograft mouse model to support the oncogenic role of CORO6 in ccRCC development. (A–C) CORO6 depletion suppressed ccRCC tumor growth monitored by tumor size (A) , growth curve (B) , and tumor weight (C) . (D) The mRNA levels of c-Myc, AXIN2, and CCND1 were remarkably reduced in CORO6-depleted ccRCC tumors. The expression levels of detected genes were normalized to 18s rRNA. (E) Western blotting showed that the protein levels of c-Myc, AXIN2, and CCND1 were remarkably reduced in CORO6-depleted ccRCC tumors. GAPDH was used as the internal control. (F–H) CORO6-promoted ccRCC tumor growth was inhibited by the administration of IWP-O1 monitored by tumor size (F) , growth curve (G) , and tumor weight (H) . ** P < 0.01, *** P < 0.001, and **** P < 0.0001. Compared with CORO6 OE, ### p < 0.001 and #### p < 0.0001.

Article Snippet: The slides were incubated with endogenous peroxidase blocking solution, and then incubated with the CORO6 (17243-1-AP, Proteintech) antibody at 4°C overnight.

Techniques: Expressing, Western Blot, Control

Journal: Journal of Cancer

Article Title: Coronin 6 promotes hepatocellular carcinoma progression by enhancing canonical Wnt/beta-catenin signaling pathway

doi: 10.7150/jca.62873

Figure Lengend Snippet: Primers for construction of plasmid in this study.

Article Snippet: Primary antibody against CORO6 (Proteintech; Catalog number: 17243-1-AP) diluted (1:100) in 1% BSA (dissolved in PBST) was used to incubate the slide overnight.

Techniques: Plasmid Preparation, Sequencing

Journal: Journal of Cancer

Article Title: Coronin 6 promotes hepatocellular carcinoma progression by enhancing canonical Wnt/beta-catenin signaling pathway

doi: 10.7150/jca.62873

Figure Lengend Snippet: RT-qPCR primers used in this study.

Article Snippet: Primary antibody against CORO6 (Proteintech; Catalog number: 17243-1-AP) diluted (1:100) in 1% BSA (dissolved in PBST) was used to incubate the slide overnight.

Techniques: Sequencing

Journal: Journal of Cancer

Article Title: Coronin 6 promotes hepatocellular carcinoma progression by enhancing canonical Wnt/beta-catenin signaling pathway

doi: 10.7150/jca.62873

Figure Lengend Snippet: CORO6 mRNA is increased during HCC progression and is correlated with poor prognosis. (A) Comparison of CORO6 mRNA level in samples from normal tissues (n=50) and primary HCC tumor tissues (n=369). The RNA-seq raw data were derived from the TCGA-LIHC database ( https://portal.gdc.cancer.gov/ ). (B - H) Comparisons of CORO6 mRNA expression in HCC tumor tissues classified by AFP level (B), Child-Pugh grade (C), histologic grade (D), TNM stage (E), T status (F), vascular invasion (G) and living status (H). Significance was calculated between indicated groups by using Mann-Whitney test. (I) Survival probability of patients stratified by low and high CORO6 mRNA levels. * P < 0.05; ** P <0.01; *** P < 0.001; NS: not significant.

Article Snippet: Primary antibody against CORO6 (Proteintech; Catalog number: 17243-1-AP) diluted (1:100) in 1% BSA (dissolved in PBST) was used to incubate the slide overnight.

Techniques: Comparison, RNA Sequencing, Derivative Assay, Expressing, MANN-WHITNEY

Journal: Journal of Cancer

Article Title: Coronin 6 promotes hepatocellular carcinoma progression by enhancing canonical Wnt/beta-catenin signaling pathway

doi: 10.7150/jca.62873

Figure Lengend Snippet: Correlation between clinicopathological variables and CORO6 expression in HCC.

Article Snippet: Primary antibody against CORO6 (Proteintech; Catalog number: 17243-1-AP) diluted (1:100) in 1% BSA (dissolved in PBST) was used to incubate the slide overnight.

Techniques: Expressing

Journal: Journal of Cancer

Article Title: Coronin 6 promotes hepatocellular carcinoma progression by enhancing canonical Wnt/beta-catenin signaling pathway

doi: 10.7150/jca.62873

Figure Lengend Snippet: CORO6 protein is upregulated in HCC specimens and cell lines. (A) Immunohistochemical (IHC) staining of CORO6 protein in adjacent normal tissues and tumor tissues on the tissue microarray. Representative images are shown. (B) Statistical comparison of CORO6 protein level in IHC-stained adjacent normal tissues (n=50) and tumor tissues (n=50). Significance analyses were performed between indicated groups by using unpaired Mann-Whitney test (left panel) and paired Wilcoxon matched-paired signed rank test (right panel), respectively. (C) Statistical analyses of CORO6 protein level in IHC-stained tumor tissues (n=50) classified by TNM stage (left panel) and T status (right panel). Significance was calculated between indicated groups with Mann-Whitney. (D) Quantification of CORO6 mRNA level in normal liver cell (control) as well as HCC cells via RT-qPCR. Significance analyses were performed between indicated groups by using multiple comparisons followed by Turkey test. (E and F) Western blotting analysis of CORO6 protein level in normal liver cell and HCC cells (E). Significance was calculated between indicated groups with multiple comparisons followed by Turkey test (F). ** P <0.01; *** < 0.001; **** P < 0.0001; ns: not significant.

Article Snippet: Primary antibody against CORO6 (Proteintech; Catalog number: 17243-1-AP) diluted (1:100) in 1% BSA (dissolved in PBST) was used to incubate the slide overnight.

Techniques: Immunohistochemical staining, Immunohistochemistry, Microarray, Comparison, Staining, MANN-WHITNEY, Control, Quantitative RT-PCR, Western Blot

Journal: Journal of Cancer

Article Title: Coronin 6 promotes hepatocellular carcinoma progression by enhancing canonical Wnt/beta-catenin signaling pathway

doi: 10.7150/jca.62873

Figure Lengend Snippet: Correlation between clinicopathological variables and CORO6 expression in HCC.

Article Snippet: Primary antibody against CORO6 (Proteintech; Catalog number: 17243-1-AP) diluted (1:100) in 1% BSA (dissolved in PBST) was used to incubate the slide overnight.

Techniques: Expressing

Journal: Journal of Cancer

Article Title: Coronin 6 promotes hepatocellular carcinoma progression by enhancing canonical Wnt/beta-catenin signaling pathway

doi: 10.7150/jca.62873

Figure Lengend Snippet: CORO6 depletion suppresses HCC cell proliferation, and promotes apoptosis. (A - C) Knockdown efficiency validation of three independent shRNAs targeting CORO6 by RT-qPCR (A) and western blotting (B) in Hep3B and SMMC-7721 cells. Significance analyses of CORO6 protein level were performed between negative control groups (shNC) and groups treated with different shRNAs targeting CORO6 (shCORO6-1, shCORO6-2 and shCORO6-3) by using unpaired Student's t-tests (C). (D) Assessment of cell viability by MTT assay upon CORO6 knockdown in Hep3B and SMMC-7721 cells. Significance was calculated between negative control groups (shNC) and groups treated with different shRNAs targeting CORO6 (shCORO6-1 and shCORO6-2) by using unpaired Student's t-tests. (E) Colony formation assay of Hep3B and SMMC-7721 cells upon CORO6 depletion. Representative images of colonies are exhibited in the upper panel. Significance was calculated between negative control groups (shNC) and groups treated with different shRNAs targeting CORO6 (shCORO6-1 and shCORO6-2) by using unpaired Student's t-tests (lower panel). (F) Cell cycle analyses of Hep3B and SMMC-7721 cells with CORO6 knockdown. Representative images of FACS analyses are exhibited in the upper panel. Significance analyses were carried out between negative control groups (shNC) and groups treated with different shRNAs targeting CORO6 (shCORO6-1 and shCORO6-2) by using unpaired Student's t-tests (lower panel). (G) Detection of cell apoptosis with Annexin V/PI staining assays in CORO6 knockdown Hep3B and SMMC-7721 cells. Representative images of FACS analyses are shown in the upper panel. Significance analyses were carried out between negative control groups (shNC) and groups treated with different shRNAs targeting CORO6 (shCORO6-1 and shCORO6-2) by using unpaired Student's t-tests (lower panel). * P < 0.05; ** P <0.01; *** P < 0.001; **** P < 0.0001.

Article Snippet: Primary antibody against CORO6 (Proteintech; Catalog number: 17243-1-AP) diluted (1:100) in 1% BSA (dissolved in PBST) was used to incubate the slide overnight.

Techniques: Knockdown, Biomarker Discovery, Quantitative RT-PCR, Western Blot, Negative Control, MTT Assay, Colony Assay, Staining

Journal: Journal of Cancer

Article Title: Coronin 6 promotes hepatocellular carcinoma progression by enhancing canonical Wnt/beta-catenin signaling pathway

doi: 10.7150/jca.62873

Figure Lengend Snippet: CORO6 depletion inhibits migratory and invasive abilities. (A) Assessment of cell migratory ability in Hep3B and SMMC-7721 cells with CORO6 depletion. Representative images are shown in the left panel. Significance was calculated between negative control groups (shNC) and groups treated with different shRNAs targeting CORO6 (shCORO6-1 and shCORO6-2) by using unpaired Student's t-tests (right panel). (B) Evaluation of cell invasive ability in Hep3B as well as SMMC-7721 cells upon CORO6 knockdown. Representative images are shown in the left panel. Significance was calculated between negative control groups (shNC) and groups treated with different shRNAs targeting CORO6 (shCORO6-1 and shCORO6-2) by using unpaired Student's t-tests (right panel). *** P < 0.001; **** P < 0.0001.

Article Snippet: Primary antibody against CORO6 (Proteintech; Catalog number: 17243-1-AP) diluted (1:100) in 1% BSA (dissolved in PBST) was used to incubate the slide overnight.

Techniques: Negative Control, Knockdown

Journal: Journal of Cancer

Article Title: Coronin 6 promotes hepatocellular carcinoma progression by enhancing canonical Wnt/beta-catenin signaling pathway

doi: 10.7150/jca.62873

Figure Lengend Snippet: CORO6 activates Wnt signaling by promoting WNT10B expression. (A) Gene set enrichment analysis (GSEA) for RNA-seq data from TCGA-LIHC database. (B) Statistical comparison of WNT1 , WNT3 and WNT10B mRNA expression between normal and tumor samples from TCGA-LIHC database by using unpaired Student's t-tests. (C) Detection of WNT10B level via RT-qPCR in Hep3B and SMMC-7721 cells with CORO6 ectopic expression by using unpaired Student's t-tests. (D) TOP-flash/FOP-flash dual luciferase assay for detecting Wnt/b-catenin-driven downstream transcriptional activity. FOP-flash activity was set as a negative control. Unpaired Student's t-tests were performed to test the statistical significance. * P < 0.05; *** P < 0.001; **** P < 0.0001; ns: not significant.

Article Snippet: Primary antibody against CORO6 (Proteintech; Catalog number: 17243-1-AP) diluted (1:100) in 1% BSA (dissolved in PBST) was used to incubate the slide overnight.

Techniques: Expressing, RNA Sequencing, Comparison, Quantitative RT-PCR, Luciferase, Activity Assay, Negative Control

Journal: Journal of Cancer

Article Title: Coronin 6 promotes hepatocellular carcinoma progression by enhancing canonical Wnt/beta-catenin signaling pathway

doi: 10.7150/jca.62873

Figure Lengend Snippet: CORO6 knockdown inhibits HCC cell growth in vivo by attenuating Wnt pathway. (A) Validation of CORO6 knockdown efficiency by qPCR in Hep3B stable cell lines. Analysis was carried out using unpaired Student's t-tests. (B and C) Confirmation of CORO6 knockdown efficiency by western blotting in Hep3B stable cell lines (B). Analysis was performed by unpaired Student's t-tests (C). (D) Representative images of tumors isolated from mice that are subjected to Hep3B cells with CORO6 depletion. At the top of the image is a scale, the unit is cm. (E and F) Quantification regarding volume of tumors (E) as well as weight (F) in mice (n = 6) that were injected with CORO6 depleted Hep3B cells. Unpaired Student's t-tests were done between the control group (shNC) and CORO6 knockdown groups (shCORO6-1 and shCORO6-2), respectively. (G and H) Western blotting assessment of protein levels of Wnt target genes in tumors formed by Hep3B stable cells with CORO6 knockdown (G). Unpaired Student's t-tests were carried out between the control group (shNC) and CORO6 knockdown groups (shCORO6-1 and shCORO6-2), respectively (H). ** P <0.01; *** P < 0.001; **** P < 0.0001.

Article Snippet: Primary antibody against CORO6 (Proteintech; Catalog number: 17243-1-AP) diluted (1:100) in 1% BSA (dissolved in PBST) was used to incubate the slide overnight.

Techniques: Knockdown, In Vivo, Biomarker Discovery, Stable Transfection, Western Blot, Isolation, Injection, Control