Dynamic changes of different phenotypic and genetic circulating tumor cells as a biomarker for evaluating the prognosis of RCC

Figures & data

Table 1. Demographic characteristics of patients with RCC.

Variable	All	Metastasis-free group	Metastatic group
Number, n (%)	69	58 (84.1%)	11 (15.9%)
Age (y)	57.5 ± 11.1	57.1 ± 11.6	59.7 ± 7.8
Gender
Male, n (%)	47 (68.1%)	40 (68.9%)	7 (63.6%)
Female, n (%)	22 (31.9%)	18 (31.1%)	4 (36.4%)
Pathological type, n (%)
Clear-cell carcinoma	59 (85.5%)	50 (86.2%)	9 (81.8%)
Papillary carcinoma	5 (7.2%)	4 (6.9%)	1 (9.1%)
Chromophobe carcinoma	4 (5.8%)	3 (5.2%)	1 (9.1%)
Eosinophilic tumor	1 (1.5%)	1 (1.7%)	0 (0%)
Pathological grading
I	8 (11.6%)	7 (12.1%)	1 (9.1%)
I-II	3 (4.3%)	3 (5.2%)	0 (0%)
II	49 (71.1%)	40 (69.0%)	9 (81.8%)
II-III	6 (8.7%)	6 (10.3%)	0 (0%)
III	3 (4.3%)	2 (3.4%)	1 (9.1%)
TNM staging
T1	41 (59.4%)	38 (65.5%)	3 (27.3%) ^a,b
T2	25 (36.2%)	18 (31.1%)	7 (63.6%) ^a,b
T3	2 (2.9%)	1 (1.7%)	1 (9.1%)
T4	1 (1.5%)	1 (1.7%)	0 (0%)
N0	68 (98.5)	58 (100%)	10 (90.9%)
N1	1 (1.5%)	0 (0%)	1 (9.1%)
M0	69 (100%)	58 (100%)	11 (100%)
M1	0 (0%)	0 (0%)	0 (0%)
Surgical treatments
Radical nephrectomy	26 (37.7%)	17 (29.3%)	9 (81.8%) ^a,b
Partial nephrectomy	43 (62.3%)	41 (70.7%)	2 (18.2%) ^a,b

Data presented as mean ± SD or n (%) and P < 0.05 was considered statistically significant.

^a P < 0.05 vs. All; ^b P < 0.05 vs. Metastasis-free group.

Table 2. The tumor diameter and renal score in the metastasis-free group and metastatic group.

Variable	Metastasis-free group	Metastatic group	P
Tumor diameter (cm)	4.64 ± 1.32	7.15 ± 1.78	0.002
Renal score	7.58 ± 1.60	9.91 ± 0.70	< 0.001
Median	8 (4–11)	10 (9–11)

Data presented as mean ± SD.

Figure 1. Three types of CTCs in kidney cancers. Left panel showed the images of epithelial, mixed and mesenchymal CTCs. Epithelial CTCs presented only Alexa Fluor 594 (Red color) labeled epithelial markers (EpCAM and CK8/18/19), mesenchymal CTCs exhibited only Alexa Fluor 488 (Green color) labeled mesenchymal markers (Vimentin and Twist), and mixed CTCs having both epithelial and mesenchymal markers were stained with both green and red immunofluorescent dyes. Right panel showed the distribution of three CTCs types in kidney cancers.

Figure 2. Initial CTCs counts in two groups before surgery.

Figure 3. Dynamic changes of three CTCs types in the metastatic group. (Data were presented as mean ± SD and P < 0.05 was considered statistically significant. Pre-op: preoperatively, post-op 6m: 6 months postoperatively, post-op 12m: 12 months postoperatively. * P < 0.05 Post-op 12m vs. Pre-op; ** P < 0.05 Post-op 12m vs. Post-op 6m.).

Figure 4. Dynamic changes of three CTCs types in the metastasis-free group. (Data were presented as mean ± SD and P < 0.05 was considered statistically significant. Pre-op: preoperatively, post-op 6m: 6 months postoperatively, post-op 12m: 12 months postoperatively. * P < 0.05 Post-op 12m vs. Pre-op; ** P < 0.05 Post-op 12m vs. Post-op 6m.).

Figure 5. Beclin-1 expression in CTCs. Epithelial markers were labeled with Alexa Fluor 594 (Red color); mesenchymal markers were labeled with Alexa Fluor 488 (Green color); Beclin-1 marker was labeled by Alexa Fluor 647 (Purple color). Right panel showed the distribution of three CTCs types in kidney cancers.

Figure 6. The expression of Beclin1 in different types of CTCs preoperatively. (Data were presented as mean ± SD and P < 0.05 was considered statistically significant. * P < 0.05 Beclin1 (+) mixed CTCs vs. Beclin1 (-) mixed CTCs; ** Beclin1 (+) all CTCs vs. Beclin1 (-) all CTCs).

Figure 7. The expression of Beclin1 in CTCs preoperatively in the metastasis-free group and metastatic group. (Data were presented as mean ± SD and P < 0.05 was considered statistically significant. * P < 0.05 Beclin1 (+) CTCs vs. Beclin1 (-) CTCs in the metastatic group).

Figure 8. Dynamic changes of Beclin1 expression in CTCs in metastatic group. (Data were presented as mean ± SD and P < 0.05 was considered statistically significant. Pre-op: preoperatively, post-op 6m: 6 months postoperatively, post-op 12m: 12 months postoperatively. * P < 0.05 Post-op 12m vs. Pre-op; ** P < 0.05 Post-op 12m vs. Post-op 6m; # P < 0.05 Post-op 6m vs. Pre-op.).