Fig. 6
Knocking down CCT3 in TRCs inhibits tumor formation. a DEG stemness enrichment analysis reveals overexpression and silence of CCT3 were both enriched in stemness-related terms. Red represents the result of CCT3 overexpression vs negative control, and green represents the result of siCCT3 vs negative control. b CCT3 promoted the SP cells ratio. Flow cytometric analyses of the SP fraction of HeLa cells overexpressed or silenced CCT3 for 24 h (right). Quantification of the SP fraction of HeLa cells (left). c Overexpression of CCT3-promoted stemness gene upregulated in HeLa 2D. d CCT3-related stemness genes shown higher expression levels of protein in TRCs. Representative western-blotting images of HeLa cells and Bel7402 cells. e Tumorigenicity of 2D cells and TRCs in BALB/c nude mice. 2D cells were cultured on rigid plastic, TRCs were cultured in 90 Pa fibrin gels for 5 days, TRCs + shCCT3 were TRCs transfected with shCCT3 for 12 h. They were then subcutaneously xenotransplanted into BALB/c nude mice with 50,000 cells per mice. The tumor formation ratios were recorded after 40 days subcutaneous injection. f, g Silence of CCT3-inhibited tumor formation ability of HeLa and Bel7402 TRCs. A total of 5 × 105 TRCs with scramble shRNA transfected were subcutaneous injected into left posterior subcutaneous of BALB/c nude mice, and TRCs transfected with CCT3 shRNA transfected were injected in the other side. Representative photos of tumor-bearing mice were shown (left), bar = 1 cm. Data are represented as the mean ± s.e.m. of n = 3 replicates (b, c). Asterisks in b are compared with HeLa NC group. *p < 0.05, **p < 0.01, ***p < 0.001; ns, not significantly different (Student’s t test)
