https://orcid.org/0000-0002-0779-0530
Figures & data
Figure 1 The overexpression of CCDC34 in HCC tissues.
Notes: (A) The expression of CCDC34 in the HCC tissues and adjacent normal tissues from TCGA database (n=50). (B) The overall survival rate of patients with high or low CCDC34 expression in TCGA database (P=0.004). (C) The qRT-PCR assays for the CCDC34’s mRNA expression in the HCC tissues and adjacent normal tissues (n=21). **P<0.01. (D) Representative Western blot assays for the CCDC34 protein expression in the HCC tissues (T) and adjacent normal tissues (N).
Abbreviations: HCC, hepatocellular carcinoma; TCGA, the Cancer Genome Atlas; qRT-PCR, quantitative real-time polymerase chain reaction.
Figure 2 HCC cell proliferation and clonogenicity in vitro facilitated by CCDC34.
Notes: (A) The knockdown efficiency of shCCDC34 in the MHCC97-H and SMMC-7721 cell lines measured by the qRT-PCR and Western blot. (B and C) The proliferation ability of the MHCC97-H and SMMC-7721 cells transfected with shCCDC34 or NC lentivirus were detected by the CCK-8 assays (B) and the colony formation assays (C). The “sh” represents the shCCDC34 group and NC stands for the negative control group. (D) The overexpression efficiency of CCDC34 in the BEL-7404 and SMMC-7721 cell lines were measured by the qRT-PCR and Western blot (E and F). The proliferation ability of the BEL-7404 and SMMC-7721 cells transfected with the CCDC34-OE or NC lentivirus were detected by the CCK-8 assays (E) and the colony formation assays (F). The CCDC34 represents the CCDC34-OE group and NC stands for the negative control group. *P<0.05; **P<0.01; ***P<0.001.
Abbreviations: HCC, hepatocellular carcinoma; qRT-PCR, quantitative real-time polymerase chain reaction; NC, negative control; CCK-8, Cell Counting Kit-8; OE, overexpression.
Figure 3 HCC cell migration and invasion in vitro regulated by CCDC34.
Notes: (A and B) The migration and invasion ability of the MHCC97-H (A) and SMMC-7721 (B) cells transfected with the shCCDC34 or NC lentivirus were measured by the transwell assays. (C and D). The migration and invasion ability of the BEL-7404 (C) and SMMC-7721 (D) cells transfected with the CCDC34-OE or NC lentivirus were measured by the transwell assays. Scale bar = 100 μm. **P<0.01; ***P<0.001.
Abbreviations: HCC, hepatocellular carcinoma; NC, negative control; OE, overexpression.
Figure 4 HCC cell growth and metastasis in vivo suppressed by CCDC34-KD.
Notes: (A) The MHCC97-H cells transfected with shCCDC34 or NC lentivirus were subcutaneously injected into the nude mice (n=8). The top tumors were produced by MHCC97-H-NC cells, while the bottom by MHCC97-H-shCCDC34 cells. (B) The volume and weight of tumors harvested from the nude mice were measured. (C) Representative H&E staining in lung sections obtained from the nude mice injected with the MHCC97-H-shCCDC34 or NC cells through the tail veins (n=7). Scale bar = 500 μm (left) and 50 μm (right). The top tumors were produced by MHCC97-H-NC cells, while the bottom by MHCC97-H-shCCDC34 cells. (D) The number of metastatic tumors per lung was counted. **P<0.01; ***P<0.001.
Abbreviations: KD, knockdown; HCC, hepatocellular carcinoma; NC, negative control.
Figure 5 The activation of AKT pathways and EMT process hampered by silencing of CCDC34.
Notes: (A) A scatter plot shows the number of the DEGs analyzed by the RNA sequencing in the BEL-7404-CCDC34-OE cells (fold change >2.0 and P-value <0.05). (B) The KEGG pathway enrichment analysis of all the DEGs. (C) A heatmap consisting of the DEGs extracted from the five HCC-related modules (red) of . (D) Protein-protein interaction (PPI) network analysis for the DEGs extracted from the five HCC-related modules (red) of . The size of the nodes indicates the number of neighboring nodes directly connected to the node. (E) Representative Western blot assays for the protein level of CCDC34, AKT, p-AKT, P21, CCND1, N-cadherin and E-cadherin.
Abbreviations: EMT, epithelial-mesenchymal transition; DEGs, differentially expressed genes; OE, overexpression; KEGG, Kyoto Encyclopedia of Genes and Genomes; PPI, protein-protein interaction.
