Chaperonin containing TCP1 subunit 6A may activate Notch and Wnt pathways to facilitate the malignant behaviors and cancer stemness in oral squamous cell carcinoma

Figures & data

Figure 1. CCT6A expression. CCT6A mRNA expression (a) and protein expression (b, c) were increased in OSCC cells compared to control cells.

Figure 2. CCT6A induced OSCC malignant behaviors. CCT6A mRNA expression (a) and protein expression (b, c) after transfection into SCC-15 cells. Effect of CCT6A on cell proliferation (d), cell apoptosis rate (e, g) and invasive cell count (f, h) in SCC-15 cells. CCT6A mRNA expression (i) and protein expression (j, k) after transfection into HSC-3 cells. Effect of CCT6A on cell proliferation (i), cell apoptosis rate (m, o) and invasive cell count (n, p) in HSC-3 cells.

Figure 3. CCT6A promoted OSCC stemness. Effect of CCT6A on sphere formatted number (a, b), CD133 and sox expressions (c, d) in SCC-15 cells. Effect of CCT6A on sphere formatted number (e, f), CD133 and sox expressions (g, h) in HSC-3 cells.

Figure 4. Cross analysis. Cross analysis (a) of accordant DEGs. Further GO enrichment (b) and KEGG enrichment (c) analyses for cross analysis.

Table 1. Top 50 accordant DEGs.

Gene ID	Symbols	OE-CCT6A vs. OE-NC				KO-CCT6A vs. KO-NC				Abs mean Log2FC
		log2FC	P value	Padj value	Trend	log2FC	P value	Padj value	Trend
ENSG00000148400	NOTCH1	−2.378411	0.000687	0.03134	DOWN	2.625648	5.97E–11	7.23E–08	UP	2.502029
ENSG00000167550	RHEBL1	−2.518068	5.56E–06	0.001249	DOWN	1.645860	2.13E–05	0.001952	UP	2.081964
ENSG00000159714	ZDHHC1	−2.461677	0.000599	0.029344	DOWN	1.355889	0.002503	0.044246	UP	1.908783
ENSG00000183431	SF3A3	−2.501450	3.92E–05	0.004572	DOWN	1.312106	0.000436	0.014909	UP	1.906778
ENSG00000169884	WNT10B	−2.347524	6.68E–09	8.75E–06	DOWN	1.432256	5.16E–06	0.000751	UP	1.88989
ENSG00000154582	TCEB1	−1.875135	2.67E–07	0.000124	DOWN	1.739005	2.79E–08	1.22E–05	UP	1.80707
ENSG00000120820	GLT8D2	−1.533041	1.09E–06	0.000416	DOWN	2.007723	3.09E–11	4.05E–08	UP	1.770382
ENSG00000165275	TRMT10B	−1.893333	5.44E–13	2.85E–09	DOWN	1.531602	2.9E–09	2.06E–06	UP	1.712468
ENSG00000140945	CDH13	−2.007222	5.32E–13	2.85E–09	DOWN	1.391215	3.32E–07	9.01E–05	UP	1.699218
ENSG00000177697	CD151	−1.798611	9.75E–05	0.008474	DOWN	1.594193	1.62E–05	0.001583	UP	1.696402
ENSG00000213465	ARL2	−1.398399	2.01E–06	0.000644	DOWN	1.986303	3.07E–12	6.89E–09	UP	1.692351
ENSG00000099942	CRKL	−1.858430	0.0003	0.020025	DOWN	1.455991	0.000321	0.012272	UP	1.65721
ENSG00000256943	RP13-895J2.2	−1.610149	2.82E–07	0.000127	DOWN	1.661801	4.06E–08	1.68E–05	UP	1.635975
ENSG00000214135	AC024560.3	−1.740457	9.31E–05	0.008184	DOWN	1.503051	8.82E–05	0.005319	UP	1.621754
ENSG00000100591	AHSA1	−1.189496	0.001214	0.042849	DOWN	1.927258	1.23E–08	6.49E–06	UP	1.558377
ENSG00000172380	GNG12	−1.845059	1.12E–05	0.002017	DOWN	1.236532	7.22E–05	0.004619	UP	1.540796
ENSG00000165511	C10orf25	−1.852807	0.00017	0.013211	DOWN	1.191650	0.001598	0.03325	UP	1.522229
ENSG00000186603	HPDL	−2.140821	1.46E–12	5.74E–09	DOWN	0.895481	0.002283	0.042028	UP	1.518151
ENSG00000139133	ALG10	−1.536813	3.07E–06	0.000819	DOWN	1.484480	3.27E–06	0.000514	UP	1.510646
ENSG00000184208	C22orf46	−1.456098	2.23E–05	0.003142	DOWN	1.512975	1.41E–06	0.000281	UP	1.484536
ENSG00000023697	DERA	−1.220654	0.000739	0.033162	DOWN	1.712880	2.6E–08	1.17E–05	UP	1.466767
ENSG00000175164	ABO	−1.590764	9.78E–08	6.08E–05	DOWN	1.318285	1.38E–06	0.000278	UP	1.454524
ENSG00000183020	AP2A2	−1.412515	0.001152	0.041975	DOWN	1.491105	3.16E–05	0.00256	UP	1.45181
ENSG00000103269	RHBDL1	−1.710446	6.14E–09	8.75E–06	DOWN	1.174036	1.15E–05	0.001253	UP	1.442241
ENSG00000070950	RAD18	−1.215714	5E–05	0.005395	DOWN	1.636560	1.25E–08	6.49E–06	UP	1.426137
ENSG00000214575	CPEB1	−1.788342	5.77E–09	8.75E–06	DOWN	1.045225	0.000232	0.01001	UP	1.416783
ENSG00000066468	FGFR2	1.159953	6.57E–05	0.006305	UP	−1.634556	6.15E–07	0.000144	DOWN	1.397254
ENSG00000203875	SNHG5	−1.883577	9.58E–10	1.67E–06	DOWN	0.907840	0.00087	0.022807	UP	1.395708
ENSG00000124802	EEF1E1	−1.521077	3.91E–05	0.004572	DOWN	1.238579	0.000293	0.011657	UP	1.379828
ENSG00000143252	SDHC	−1.436719	0.001178	0.042506	DOWN	1.321061	0.000614	0.018645	UP	1.37889
ENSG00000148634	HERC4	−1.648579	2.01E–07	0.000102	DOWN	1.097867	0.000424	0.014839	UP	1.373223
ENSG00000173334	TRIB1	−1.279918	0.001206	0.042846	DOWN	1.446426	3.96E–05	0.003017	UP	1.363172
ENSG00000100426	ZBED4	−1.278577	1.05E–05	0.001949	DOWN	1.442516	5.86E–07	0.00014	UP	1.360547
ENSG00000170579	DLGAP1	−1.476191	0.000112	0.00942	DOWN	1.242064	0.000271	0.011093	UP	1.359127
ENSG00000196754	S100A2	0.944327	0.000901	0.036548	UP	−1.768450	5.7E–10	5.97E–07	DOWN	1.356388
ENSG00000074621	SLC24A1	−1.526180	3.52E–05	0.004332	DOWN	1.177001	7.97E–05	0.004936	UP	1.35159
ENSG00000131067	GGT7	−1.420325	0.000429	0.024702	DOWN	1.275687	5.9E–05	0.003933	UP	1.348006
ENSG00000278341	RP5-1142A6.10	−1.181115	3.8E–05	0.004529	DOWN	1.487821	1.72E–07	5.43E–05	UP	1.334468
ENSG00000157368	IL34	−1.347643	2.24E–05	0.003142	DOWN	1.299019	1.14E–05	0.001244	UP	1.323331
ENSG00000084774	CAD	−1.589267	1.4E–08	1.45E–05	DOWN	1.047985	0.000152	0.007652	UP	1.318626
ENSG00000168214	RBPJ	1.024709	9.46E–05	0.008269	UP	−1.604224	1.64E–09	1.46E–06	DOWN	1.314466
ENSG00000162552	WNT4	1.041606	0.000111	0.00942	UP	−1.566537	7.78E–09	4.71E–06	DOWN	1.304072
ENSG00000175029	CTBP2	−1.583170	1.14E–06	0.000416	DOWN	1.010773	0.000169	0.008148	UP	1.296971
ENSG00000258144	RP11-554L12.1	−1.557405	2.13E–07	0.000105	DOWN	1.034771	0.000179	0.008432	UP	1.296088
ENSG00000153037	SRP19	−1.686000	4.63E–08	3.47E–05	DOWN	0.904008	0.000692	0.019813	UP	1.295004
ENSG00000247271	ZBED5-AS1	−0.964932	0.001375	0.046729	DOWN	1.601315	3.45E–09	2.26E–06	UP	1.283123
ENSG00000107175	CREB3	1.106838	3.99E–06	0.000996	UP	−1.450459	2.57E–09	2.02E–06	DOWN	1.278649
ENSG00000275854	RP11-278C7.5	−1.469965	1.77E–06	0.000604	DOWN	1.083279	2.27E–05	0.002019	UP	1.276622
ENSG00000111786	SRSF9	−1.307128	0.000233	0.016756	DOWN	1.225859	0.000142	0.007448	UP	1.266494
ENSG00000106948	AKNA	−1.118108	0.001215	0.042849	DOWN	1.410606	2.07E–06	0.000379	UP	1.264357

Top 50 DEGs were selected by the rank of absolute mean value of Log2FC. DEGs, differentially expressed genes; ID, identification; FC, fold change; adj, adjust; abs, absolute; OE, overexpression; KO, knockout; NC, negative control.

Table 2. Top 5 pathways.

Pathways	Num of symbols	Proportion of symbols	Symbols	Fold enrichment	P value
Notch signaling pathway	10	0.172	DLL3, DTX2, CTBP2, HDAC1, HDAC2, RBPJ, JAG2, NOTCH1, KAT2B, CIR1	25.406	8.00161E–11
Basal cell carcinoma	6	0.103	WNT4, WNT10B, WNT2B, WNT9A, FZD8, FZD9	13.303	4.81089E–08
Wnt signaling pathway	10	0.172	CTBP2, DAAM1, WNT4, CHD8, SFRP1, WNT10B, WNT2B, WNT9A, FZD8, FZD9	8.528	1.63219E–06
Signaling pathways regulating pluripotency of stem cells	9	0.155	FGFR2, PIK3CA, PIK3CD, WNT4, WNT10B, WNT2B, WNT9A, FZD8, FZD9	7.729	1.48293E–05
Chronic myeloid leukemia	7	0.121	CRKL, CTBP2, HDAC1, HDAC2, MDM2, PIK3CA, PIK3CD	11.694	2.17139E–05

These pathways were enriched by KEGG enrichment in accordance DEGs and displayed by the rank of P values. KEGG, Kyoko Encyclopedia of Genes and Genomes; DEGs, differentially expressed genes.

Figure 5. CCT6A activated Wnt and Notch pathways in OSCC. Effect of CCT6A on Wnt and Notch pathways in SCC-15 cells (a) and in HSC-3 cells (b).

Figure 6. Notch1 and Wnt4 compensated the effect of CCT6A on regulating Notch and Wnt pathways. CCT6A mRNA expression (a) Notch1 mRNA expression (b), Wnt4 mRNA expression (c) and their protein expressions (d, e) in rescue experiments.

Figure 7. Notch1 and Wnt4 compensated the effect of CCT6A knockout on regulating OSCC malignant behaviors. Notch1 and Wnt4 attenuated the effect of CCT6A knockout on regulating cell proliferation (a), cell apoptosis rate (b, d) and invasive cell count (c, e) in SCC-15 cells.

Figure 8. Notch1 and Wnt4 compensated the effect of CCT6A knockout on regulating OSCC stemness. Notch1 and Wnt4 attenuated the effect of CCT6A knockout on regulating sphere formatted number (a, b), CD133 and sox expressions (c, d) in SCC-15 cells.

Data availability statement

The data used to support the findings of this study have been included in this article. The datasets and materials used in this study are available from the corresponding author upon reasonable request.