CCT6B as a candidate chaperonin-related molecule in hepatocellular carcinoma: Bioinformatic analysis and preliminary...

Hepatocellular carcinoma (HCC) is an aggressive malignancy with limited therapeutic options and poor prognosis, highlighting the need for prognostic biomarkers and novel molecular candidates. This study aimed to identify chaperonin-related genes associated with HCC prognosis and to preliminarily explore the biological relevance of CCT6B.

The Cancer Genome Atlas (TCGA) database and R were used to identify molecular chaperonin-related genes associated with HCC prognosis. A prognostic model was constructed using least absolute shrinkage and selection operator (LASSO)-Cox regression, and the association between CCT6B expression and survival was evaluated. Western blotting (WB) was performed to compare CCT6B protein levels in paired HCC tumor and adjacent nontumor tissues. shRNA-mediated CCT6B knockdown was conducted in SNU-182 cells, and migration, proliferation, and apoptosis were assessed using wound-healing, MTT, and flow cytometry assays, respectively. Apoptosis-related proteins (BAX, Bcl-2, Cyt-c, Caspase-9, Caspase-3, and P53) were measured by WB.

Bioinformatics analysis identified nine chaperonin-related genes associated with HCC prognosis in the TCGA-LIHC cohort. CCT6B was selected for exploratory validation due to its transcriptomic pattern and limited prior characterization in HCC. Although CCT6B mRNA was downregulated in public transcriptomic datasets, Western blot analysis of three paired clinical samples suggested a possible increase in CCT6B protein abundance in tumor tissues; however, this observation should be interpreted as preliminary because of the very small sample size. In SNU-182 cells, shRNA-mediated CCT6B knockdown was associated with reduced proliferation and migration and increased apoptosis, accompanied by changes in intrinsic apoptosis-related proteins. Because these functional data were generated in a single cell line using one knockdown strategy, they should be regarded as preliminary and hypothesis-generating.

This study identified a TCGA-derived chaperonin-related prognostic signature in HCC and prioritized CCT6B as a biologically distinctive candidate for preliminary validation. CCT6B showed transcriptomic downregulation in public datasets, while limited protein-level observations suggested a possible discordant trend that requires confirmation. CCT6B knockdown in one HCC cell line was associated with altered proliferation, migration, apoptosis, and apoptosis-related protein expression. However, because of the limited clinical sample size, single-cell-line validation, lack of rescue and gain-of-function experiments, absence of in vivo data, and unresolved mRNA-protein discordance, CCT6B should currently be regarded as a candidate chaperonin-related molecule of biological interest rather than a validated prognostic biomarker or therapeutic target in HCC.