A study on UHPLC-MS/MS analyses of DNA and RNA oxidative damage metabolites in patients with cervical carcinoma: 8-oxoG in urine as a potential biomarker of cervical carcinoma

Abstract

Objective: The purpose of this study was to compare the levels of 8-oxoG and 8-oxodG in urine of patients with cervical carcinoma and healthy women to evaluate their influences on cervical carcinoma.

Methods: In this study, urine samples were collected from 70 patients with cervical carcinoma, 24 patients with one-year follow-up, and 100 healthy women. The contents of 8-oxodG and 8-oxoG in urine were assayed by ultra-high performance liquid chromatography tandem mass spectrometry (UHPLC-MS/MS).

Results: The levels of 8-oxoG and 8-oxodG were higher in patients with cervical carcinoma (P < 0.000), while AUC of 8-oxoG and 8-oxodG was higher than 0.7. Specifically, the high-risk HPV (HR-HPV) positive group had higher 8-oxoG levels (P < 0.000), but there was no difference in 8-oxodG levels. Yet, 8-oxoG level was associated with lymphatic metastasis, lymph-vascular space infiltration (LVSI) and stromal infiltration, while 8-oxodG level was affected by the differentiation degree and stromal infiltration. According to statistics, the distinct cut-off index of lymphatic metastasis was 7.282 nmol/mmol creatinine. After operation, the concentrations of 8-oxoG and 8-oxodG dropped significantly (8-oxoG P < 0.000, 8-oxodG P = 0.004). Except for chemotherapy group, the urinary 8-oxoG dose of all treatment groups and 8-oxodG dose of chemo-radiotherapy group declined obviously.

Conclusions: 8-oxoG may be a potential biomarker for cervical carcinoma.

Keywords: 8-oxoG; Biomarker; Cervical carcinoma; Oxidative damage; UHPLC- MS/MS; Urine.

Figure 1

The concentrations of 8-oxodG and 8-oxoG in the study groups by UHPLC-MS/MS. (A) Comparison of 8-oxoG between control group and cervical carcinoma group. (B) Comparison of 8-oxodG between control group and cervical carcinoma group. (C) ROC curve of 8-oxoG in cervical carcinoma group and control group. (D) ROC curve of 8-oxodG in cervical carcinoma group and control group. (E) Comparison of 8-oxoG between HPV negative group and HPV positive group. (F) Comparison of 8-oxodG between HPV negative group and HPV positive group. (G) Comparison of 8-oxoG between HPV 16/18 group and other high-risk HPV groups. (H) Comparison of 8-oxodG between HPV 16/18 group and other high-risk HPV groups. Data were expressed as mean ± SD. ∗P < 0.05, ∗∗∗P < 0.000 were considered to be statistically significant.

Figure 2

Different clinical characteristics of 8-oxoG and 8-oxodG in cervical carcinoma group. (A) Comparison of 8-oxoG between Stage I and Stage II. (B) Comparison of 8-oxoG between small tumor group and large tumor group. (C) 8-oxoG of lymph-vascular space infiltration between negative group and positive group. (D) Expression of 8-oxoG in different histopathological types. (E) Expression of 8-oxoG in different differentiation degrees. (F) Expression of 8-oxoG in different stromal infiltration groups. (G) Comparison of 8-oxodG between Stage I and Stage II. (H) Comparison of 8-oxodG between small tumor group and large tumor group. (I) 8-oxodG of lymph-vascular space infiltration between negative group and positive group. (J) Expression of 8-oxodG in different histopathological types. (K) Expression of 8-oxodG in different differentiation degrees. (L) Expression of 8-oxodG in different stromal infiltration groups. Data were expressed as mean ± SD. ∗P < 0.05 was considered to be statistically significant.

Figure 3

Effects of 8-oxoG and 8-oxodG on lymphatic metastasis in cervical carcinoma group. (A) Comparison of 8-oxodG on lymphatic metastasis between negative group and positive group. (B) Comparison of 8-oxoG on lymphatic metastasis between negative group and positive group. (C) ROC curve of 8-oxoG on lymphatic metastasis between negative group and positive group. The cut-off value was 7.282. Data were expressed as mean ± SD. ∗P < 0.05 was considered to be statistically significant.

Figure 4

Changes of 8-oxoG and 8-oxodG in cervical carcinoma group according to different postoperative treatment methods. (A) 8-oxoG in 24 patients. (B) 8-oxoG in observation group. (C) 8-oxoG in radiotherapy group. (D) 8-oxoG in chemotherapy group. (E) 8-oxoG in chemo-radiotherapy group. (F) 8-oxodG in 24 patients. (G) 8-oxodG in observation group. (H) 8-oxodG in radiotherapy group. (I) 8-oxodG in chemotherapy group. (J) 8-oxodG in chemo-radiotherapy group. Data were expressed as mean ± SD. ∗P < 0.05, ∗∗∗P < 0.000 were considered to be statistically significant.