circPVT1 Inhibits the Proliferation and Aids in Prediction of the Prognosis of Bladder Cancer

Figures & data

Table 1 Correlation Between circPVT1 Expression and Clinicopathological Characteristics in Bladder Cancer

Characteristics	circPVT1
	No.	Low	High	P value
Age				0.056
<=60	91	22	69
>60	71	28	43
Gender				0.452
Female	95	32	63
Male	67	18	49
T Stage				0.339
T1-T2	82	22	60
T3-T4	80	28	52
N Stage				0.073
N0	69	27	42
N1-N3	93	23	70
M Stage				0.968
M0	79	25	54
M1	83	25	58
TNM Stage				0.003
I–II	111	43	68
III–IV	51	7	44
Differentiation				0.395
Poorly/moderately	81	28	53
Well	81	22	59
Tumor Size				0.310
<=4 cm	73	26	47
>4 cm	89	24	65
HER2				0.003
No mutation	112	69	43
Mutation	50	43	7

Figure 1 The expression of mRNA PVT1 and circPVT1 in BLCA and normal tissues. (A–E) Statics analysis of expression level. (A) Difference in PVT1 mRNA expression between tumor and normal tissues. (B) Difference in circPVT1 expression between tumor and normal tissues. (C) Association between mRNA PVT1 and circPVT1 expression in tumor and normal tissues. (D) Difference in PVT1 mRNA expression between tumor and normal cell lines. (E) Difference in circPVT1 expression between tumor and normal cell lines. (F and G) Correlation between mRNA PVT1 and circPVT1 expression in normal tissues (F) and BLCA tissues (G). (H) AUC: area under the ROC (receiver operating characteristic) curve of circPVT1 and mRNA PVT1. (I and J) Stability of circPVT1 and mRNA PVT1 after RNase R treatment in T24 (I) and UMUC3 (J) cell lines. ^#Represents P > 0.05, *Represents P < 0.05, **Represents P < 0.01, ***Represents P < 0.001.

Table 2 Univariate and Multivariate Cox Regression Analyses of Prognostic Factors in Bladder Cancer

Characteristics	Univariate Analysis		Multivariate Analysis
	HR (95% CI)	P value	HR (95% CI)	P value
Age (>60 vs <=60)	0.788 (0.548–1.132)	0.197
Gender (Female vs Male)	1.349 (0.946–1.923)	0.098
T Stage (T3-T4 vs T1-T2)	0.726 (0.505–1.042)	0.082
N Stage (N1-N3 vs N0)	1 (0.701–1.427)	1
TNM Stage (III–IV vs I–II)	1.6 (1.11–2.306)	0.012	0.985 (0.6–1.617)	0.953
Differentiation (Poorly/moderately vs Well)	1.186 (0.824–1.708)	0.36
Tumor Size (>4 cm vs <=4 cm)	1.257 (0.867–1.824)	0.227
HER2	1.558 (1.049–2.315)	0.028	1.284 (0.804–2.052)	0.295
circPVT1 (High vs Low)	0.406 (0.269–0.613)	< 0.001	0.443 (0.267–0.735)	0.002

Figure 2 Association of circPVT1 expression and HER2 amplification with overall survival. The representative images of HER2 negative (A) and positive (B) amplification at the DNA level. The representative images of HER2 negative (C) and positive (D) amplification by IHC at the protein level. Kaplan–Meier curves were generated by HER2 amplification (E), circPVT1 expression (F) and HER2 amplification with circPVT1 expression (G), respectively.

Figure 3 The nomogram was constructed based on circPVT1 expression and clinical characteristics. The alignment diagram of nomogram for predicting OS (A). The efficiency of the nomogram for the 3-year (B) or 5-year survival (C).

Figure 4 Knockdown of circ-PVT1 promotes BLCA proliferation in vitro and in vivo. (A–C) The proliferation in vitro and in vivo, growth curves of cell lines (A), colony formation (B), mouse tumor model (C). (D) The expression by qRT-PCR method. ^#P>0.05, *P<0.05, **P<0.01 and ***P<0.001.

Abbreviations: ERBB2, Erb-B2 Receptor Tyrosine Kinase 2; MKI67, Marker of Proliferation Ki-67; TP53, Tumor Protein P53; PCNA, Proliferating Cell Nuclear Antigen; CCND1, Cyclin D1; CCNB1, Cyclin B1.