Septin9 DNA methylation as a promising biomarker for cervical cancer

Figures & data

Table 1. Comparison of Septin9 methylation with different severity of cervical lesions.

Histological category	Methylation scores median ($\mathrm{P}\frac{1}{4}-\mathrm{P}\frac{3}{4}$)	Frequency of Septin9 methylation (%)	χ^2	p*
Cervical cancer	14.07 (5.52–29.32)	95.52 (64/67)
HSIL^a	1.77 × 10^–2 (4.00 × 10^–4–3.33)	48.00 (36/75)	95.31	1.59 × 10^–20
LSIL^b,d	1.40 × 10^–3 (3.00 × 10^–4–1.52 × 10^–2)	25.00 (13/52)
No-CIN^c,e,f	1.00 × 10^–3 (4.00 × 10^–4–6.20 × 10^–3)	14.81 (8/54)

Septin9 methylation scores were calculated using the following formula: 2[Ct (ACTB) – Ct (Septin9)] × 100.

Frequency of Septin9 methylation were used to calculate the statistical difference:

p* < 0.0001, Pearson Chi-square test.

^aCompared with cervical cancer, χ² = 38.37, P = 5.86 × 10^–10 < 0.0001, Pearson Chi-square test.

^bCompared with cervical cancer, χ² = 63.76, P = 1.41 × 10^–15 < 0.0001, Pearson Chi-square test.

^cCompared with cervical cancer, χ² = 80.83, P = 2.46 × 10^–19 < 0.0001, Pearson Chi-square test.

^dCompared with HSIL, χ² = 6.86, P = 8.84 × 10^–3 < 0.05, Pearson Chi-square test.

^eCompared with HSIL, χ² = 15.38, P = 8.80 × 10^–5 < 0.0001, Pearson Chi-square test.

^fCompared with LSIL, χ² = 1.730, P = 0.188 > 0.05, Pearson Chi-square test.

P: percentiles.

Figure 1. Methylation scores median of Septin9 with increasing severity of cervical lesions.

In this Box-and-Whisker plot, the median lines represent median values; the upper and lower lines represent the 25th and 75th percentiles, respectively.

Figure 2. The diagnostic power of Septin9 methylation, cytology and HPV16/18 genotyping.

(A) ROC of Septin9 methylation, cytology, HPV16/18 genotyping and Septin9 methylation＋HPV16/18 in differentiating cervical cancer patients from non-cancerous patients.

(B) ROC of Septin9 methylation, cytology, HPV16/18 genotyping and Septin9 methylation＋HPV16/18 in differentiating ≥ HSIL patients from ≤ LSIL patients.

Table 2. Power of Septin9 methylation, cytology and HPV16/18 genotypin for detection ≥ HSIL and cervical cancer.

Triage	Endpoint	AUC (95%CI)	Sensitivity (%) (95% CI)	Specificity (%) (95%CI)	PPV (%) (95%CI)	NPV (%) (95%CI)	YI (%)
Septin9 methylation	≥ HSIL	0.80 (0.74–0.86)	70.42 (0.63–0.78)	80.19 (0.73–0.88)	82.64 (0.75–0.88)	66.93 (0.58–0.75)	50.61
	Cervical cancer	0.92 (0.89–0.96)	95.52 (0.91–1.00)	68.51 (0.62–0.75)	52.89 (0.44–0.62)	97.64 (0.93–0.99)	64.03
Cytology (≥ASC-US)	≥ HSIL	0.71^a (0.64–0.78)	90.14^c (0.85–0.95)	51.89^e (0.42–0.61)	71.51 (0.65–0.78)	79.71 (0.69–0.88)	42.03
	Cervical cancer	0.67^b (0.60–0.74)	97.01** (0.93–1.01)	37.02^f (0.30–0.44)	36.31 (0.30–0.44)	97.10 (0.90–0.99)	34.03
HPV16/18 genotyping	≥ HSIL	0.68^a (0.61–0.75)	62.68* (0.55–0.71)	73.58*** (0.65–0.82)	76.07 (0.68–0.83)	59.54 (50.98–67.56)	36.26
	Cervical cancer	0.74^b (0.67–0.81)	82.09^d (72.91–91.27)	65.75**** (0.59–0.73)	47.01 (0.38–0.56)	90.84 (0.85–0.95)	47.83
HPV16/18 or Septin9 methylation	≥ HSIL	0.71^a (0.65–0.78)	82.39^c (0.76–0.89)	60.38^e (0.51–0.70)	73.58 (0.66–0.80)	71.91 (0.62–0.80)	42.77
	Cervical cancer	0.74^b (0.68–0.80)	98.51** (0.96–1.01)	48.62^f (0.41–0.56)	41.51 (0.34–0.49)	98.88 (0.94–1.00)	47.13

AUC: area under curve; YI: Youden index.

*Compared with the sensitivity of Septin9 methylation in group of ≥ HSIL, p > 0.05.

**Compared with the sensitivity of Septin9 methylation in group of cervical cancer, p > 0.05.

***Compared with the specificity of Septin9 methylation in group of ≥ HSIL, p > 0.05.

****Compared with the specificity of Septin9 methylation in group of cervical cancer, p > 0.05.

^aCompared with the AUC of Septin9 methylation in group of ≥ HSIL, p < 0.05.

^bCompared with the AUC of Septin9 methylation in group of cervical cancer, p > 0.05.

^cCompared with the sensitivity of Septin9 methylation in group of ≥ HSIL, p > 0.05.

^dCompared with the sensitivity of Septin9 methylation in group of cervical cancer, p > 0.05.

^eCompared with the specificity of Septin9 methylation in group of ≥ HSIL, p > 0.05.

^fCompared with the specificity of Septin9 methylation in group of cervical cancer, p > 0.05.

Table 3. Power of Septin9 methylation in plasma and SCC-Ag for prediction of pelvic nodal metastasis.

Category	AUC (95%CI)	Sensitivity (%) (95% CI)	Specificity (%) (95% CI)	PPV (%) (95% CI)	NPV (%) (95% CI)
Septin9 methylation (yes or no)	0.66 (0.54–0.78)	50.00 (0.33–0.67)	81.48 (0.73–0.90)	51.61 (0.35–0.68)	80.49 (0.71–0.88)
SCC-Ag (positive or negative)	0.56* (0.45–0.68)	65.63** (0.49–0.82)	46.91**** (0.36–0.58)	32.81 (0.23–0.45)	77.55 (0.64–0.87)
SCC-Ag or Septin9 methylation	0.61* (0.50–0.72)	78.13*** (0.64–0.92)	43.21**** (0.32–0.54)	35.21 (0.25–0.47)	83.33 (0.70–0.92)

*Compared with the AUC of Septin9 methylation, p > 0.05.

**Compared with the sensitivity of Septin9 methylation, p > 0.05.

***Compared with the sensitivity of Septin9 methylation, P > 0.05.

^****Compared with the specificity of Septin9 methylation, P > 0.05.

Data availability statement

According to Norwegian data legislation, the data of this study cannot be made generally available. Requests should be sent to the corresponding author.