Combined overexpression of HIVEP3 and SOX9 predicts unfavorable biochemical recurrence-free survival in patients with prostate cancer

Figures & data

Table 1 Clinical features of all patients

Sample type and clinical features	Experiment type (cases)
	qRT-PCR	IHC
Prostate cancer	26	98
Mean age (years)
<60	20	62
≥60	6	36
Serum PSA levels (ng/mL)
<4	5	16
≥4	21	82
Gleason score
<8	24	87
≥8	2	11
Clinical stage
<T2A	17	59
≥T2A	9	39
Metastasis	0	5
Adjacent benign prostate tissue	26	26

Abbreviations: qRT-PCR, quantitative reverse transcription polymerase chain reaction; IHC, immunohistochemistry; PSA, prostate-specific antigen.

Figure 1 Downregulation of SOX9 expression by siRNA interference inhibits HIVEP3 expression in PCa cells.

Notes: (A) Electrophoresis of qRT-PCR production. (B) Relative expression of HIVEP3 and SOX9 in PCa cells.
Abbreviations: PCa, prostate cancer; qRT-PCR, quantitative reverse transcription polymerase chain reaction; siRNA, small interfering RNA; siSOX9, siRNA for SOX9; siNC, normal control siRNA.

Figure 2 HIVEP3 and SOX9 mRNA expression in PCa and adjacent benign prostate tissues.

Notes: (A) Electrophoresis of qRT-PCR production. (B) Relative expression of HIVEP3 and SOX9 in tissue.
Abbreviations: mRNA, messenger RNA; qRT-PCR, quantitative reverse transcription polymerase chain reaction; PCa, prostate cancer.

Figure 3 Immunohistochemical staining for HIVEP3 and SOX9 proteins in PCa and adjacent benign prostate tissues.

Notes: (A) HIVEP3 strongly positive staining was found in the cytoplasm and nucleus of the PCa cell; (B) HIVEP3 weakly positive staining in the PCa was found in the cytoplasm and the nucleus of adjacent benign prostate gland cell; (C) SOX9 strongly positive staining was found in the cytoplasm and the nucleus of the PCa cell; (D) SOX9 moderately positive staining in the PCa was found in the cytoplasm and the nucleus of the adjacent benign prostate gland cell; (E) Negative immunostaining of negative controls with the primary antibody omitted in the PCa tissues; (F) Immunoreactivity scores (IRS) of SOX9 and HIVEP3 in PCa and adjacent benign prostate tissues. Inset areas in A–E is 400X magnification. The arrows in A–D show positive staining cells. **P<0.001 in comparison between PCa and adjacent benign prostate tissues.
Abbreviations: PCa, prostate cancer; IRS, immunoreactivity scores.

Table 2 Correlation between HIVEP3 and SOX9 expression in prostate cancer samples

HIVEP3 expression	SOX9 expression		rs	P-value
	High (49)	Low (49)
High (49)	37	12	0.51	<0.001
Low (49)	12	37

Abbreviation: rs, Spearman correlation coefficient.

Table 3 Association of HIVEP3 and SOX9 protein expression with the conventional clinicopathological characteristics in 98 patients with prostate cancer

Clinical features	Case number	HIVEP3		SOX9
		$\overline{X}\pm \text{s}$	P-value	$\overline{X}\pm \text{s}$	P-value
Age (years)
<60	62	3.63±0.77	0.952	7.32±0.98	0.528
≥60	36	3.64±0.83		7.19±1.00
Serum PSA Levels (ng/mL)
<4	16	3.82±0.92	0.318	7.52±1.08	0.293
≥4	82	3.60±0.76		7.24±0.96
Gleason score
<8	86	3.66±0.79	0.189	7.13±0.67	0.045
≥8	12	3.33±0.72		8.26±0.82
Clinical stage
<T2A	59	3.53±0.77	0.110	6.64±0.71	0.012
≥T2A	39	3.79±0.80		7.98±0.88
Metastasis
No	93	3.64±0.79	0.494	7.29±0.97	0.573
Yes	5	3.39±0.86		6.91 ±1.38
PSA failure
No	80	3.41±0.41	0.042	7.08±0.97	0.001
Yes	18	5.51±0.38		8.31 ±0.79

Note: Bold P-values are statistically significant.

Abbreviation: PSA, prostate-specific antigen.

Table 4 Association of combined expression of HIVEP3 and SOX9 protein with the conventional clinicopathological characteristics in 98 patients with prostate cancer

Clinical features	Case number	HIVEP3 and SOX9 expression (n)			P-value
		HIVEP3(−) SOX9(−)	HIVEP3(+) SOX9(+)	HIVEP3(+) SOX9(−)/HIVEP3(−) SOX9(+)
Age (years)
<60	60	23	20	17	0.490
≥60	38	15	16	7
Serum PSA levels (ng/mL)
<4	12	5	4	3	0.938
≥4	86	32	33	21
Gleason score
<8	87	35	31	21	0.329
≥8	11	2	6	3
Clinical stage
<T2A	59	19	28	12	0.051
≥T2A	39	18	9	12
Metastasis
No	93	35	34	24	0.370
Yes	5	2	3	0
PSA failure
No	80	35	26	19	0.024
Yes	18	2	11	5

Note: Bold P-values are statistically significant.

Abbreviation: PSA, prostate-specific antigen.

Figure 4 Kaplan–Meier survival curves of the biochemical recurrence-free survival and overall survival.

Notes: Represents the Kaplan–Meier survival curves of the biochemical recurrence-free survival and overall survival for HIVEP3 (A–C, respectively), SOX9 (D–F, respectively), combined HIVEP3 and SOX9 (G–I, respectively) expression in PCa.
Abbreviation: PCa, prostate cancer.

Table 5 Univariate and multivariate analyses of different prognostic factors in 98 patients with prostate cancer

	Hazard ratio (95% CI)	P-value
Univariate
HIVEP3	1.34 (0.08–2.37)	0.84
SOX9	0.15 (0.03–0.84)	0.032
HIVEP3/SOX9	2.58 (1.36–4.96)	<0.001
Gleason score	2.27 (1.22–4.24)	0.010
Preoperative PSA	1.02 (1.00–1.04)	0.044
Clinical stage	0.87 (0.27–2.79)	0.82
Age	0.95 (0.88–1.02)	0.15
Multivariate
SOX9	0.28 (0.08–0.91)	0.034
HIVEP3/SOX9	1.86 (1.01–3.62)	0.008
Gleason score	2.64 (1.61–4.32)	<0.001
Preoperative PSA	1.02 (1.00–1.04)	0.02

Note: Bold P-values are statistically significant.

Abbreviations: CI, confidence interval; PSA, prostate-specific antigen.