Promoter methylation of SEPT9 as a potential biomarker for early detection of cervical cancer and its overexpression predicts radioresistance

Abstract

Background Cervical cancer screening by combined cytology and HPV test has reduced the incidence of cervical cancer, but cytological screening lacks a higher sensitivity while HPV testing possesses a lower specificity. Most patients with invasive cervical cancer are treated with radiotherapy. However, insensitivity to radiotherapy leads to poor efficacy. Methods Illumina Methylation EPIC 850k Beadchip was used for genomic screening. We detected methylation of SEPT9 and mRNA expression in different cervical tissues by using methylation-specific PCR and qRT-PCR. Then using CCK8, migration assay, and flow cytometry to detect the biological function and irradiation resistance of SEPT9 in vitro and in vivo. Liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS) and co-immunoprecipitation (CoIP) were used to find the interacting gene with SEPT9. Immunostaining of CD206 in cervical cancer and polarization of macrophages (M2) were evaluated by immunofluorescence and WB. The Cancer Genome Atlas (TCGA) database was used for screening the potential miRNAs induced by SEPT9. Results Hyper-methylation of SEPT9 detects cervical cancer and normal tissues, normal+CIN1 and CIN2+CIN3+cancer with high sensitivity and specificity (AUC = 0.854 and 0.797, respectively, P < 0.001). The mRNA and protein expression of SEPT9 was upregulated in cervical cancer tissues when compared to para-carcinoma tissues. SEPT9 promotes proliferation, invasion, migration, and influences the cell cycle of cervical cancer. SEPT9 interacted with HMGB1-RB axis increases irradiation resistance. Furthermore, SEPT9 mediated miR-375 via the tumor-associated macrophages (TAMs) polarization, affecting the resistance to radiotherapy in cervical cancer. Conclusions These findings give us the evidence that SEPT9 methylation could be a biomarker for cervical cancer diagnoses. It promotes tumorigenesis and radioresistance of cervical cancer by targeting HMGB1-RB axis and causes polarization of macrophages by mediating miR-375. We suggest SEPT9 could be a potential screening and therapeutic biomarker for cervical cancer. Electronic supplementary material The online version of this article (10.1186/s13148-019-0719-9) contains supplementary material, which is available to authorized users.

Methylated Septin9: biomarker for diagnosis of cervical cancer in cervical scrapings and for prediction of pelvic lymphatic metastasis in plasma

“…In our pilot study, we also found Septin9 transcript isoform v2 was highly methylated in cervical cancer tissues, whereas no methylation in cervicitis controls. We further confirmed that in cervical scrapings, increases in Septin9 methylation were significantly correlated with greater severity of cervical lesions, which was consistent with Jiao's recent report [20]. Strangely, approximately 15% Septin9 methylation were detected in the no-CIN lesion group of scrapings, while no Septin9 methylation was detected in fresh cervicitis tissue.…”

“…For example, Connolly et al proved that methylated Septin9-v3 was associated with the occurrence of breast cancer [15], and other evidences showed Septin9-v2 also to be a novel biomarker for the detection of breast cancer [26]. A very recent study found that Septin9 was hypermethylated in cervical cancer tissues compared to the normal controls, with satisfatory sensitivity and specificity [20]. However, the feasibility of detecting Septin9 methylation in cervical scrapings has not yet been carried out.…”

“…Methylated Septin9 in circulating tumor DNA (ctDNA) has been widely utilized in colorectal cancer screening, diagnosis, and monitoring [12]. In addition, methylated Septin9 has proven to be a potential biomarker for several other tumors including gastroenteric cancer [13][14], breast cancer [15], lung cancer [16], liver cancer [17], head and neck squamous cell carcinoma [18], pancreatic cancer [19], as well as cervical cancer [20], indicating Septin9 methylation is not tissue specific. Therefore, whether ctDNA-based methylation analysis of Septin9 may provide a non-invasive approach to predicting pelvic nodal metastasis in cervical cancer?…”

The combination of Radix Astragali and Radix Angelicae Sinensis attenuates the IFN-γ-induced immune destruction of hematopoiesis in bone marrow cells

“…Medium (90 μL) and CCK-8 (10 μL) were subsequently added to each well and incubated for 2 h at 37°C. The optical density (OD value) of the cells was measured by a microplate reader (Model 3550 Microplate Reader, Bio-Rad Laboratories, Inc., Hercules, CA, USA) at a wavelength of 570 nm [25]. All experiments were performed in triplicate.…”

<p>The Regulatory Role of circRNA_101308 in Cervical Cancer and the Prediction of Its Mechanism</p>

“…3,4 Therefore, improving our understanding of the underlying cellular basis of CC development is critical for improving CC prevention and treatment. In recent decades, numerous factors, including high-risk human papillomavirus (HPV) infection, 5 immune system disorders, 6 aberrant DNA methylation, 7 and noncoding RNA deregulation, have been found to be involved in the development and progression of CC.…”