Performance Evaluation of SHOX2 and RASSF1A Methylation for the Aid in Diagnosis of Lung Cancer Based on the Analysis of FFPE Specimen

Juanhong Shi; Xue Chen; Long Zhang; Xia Fang; Yuting Liu; Xuyou Zhu; Haoyang Zhang; Lichao Fan; Jun Gu; Suxia Zhang; Bin She; Hongxiu Han; Xianghua Yi

doi:10.3389/fonc.2020.565780

Performance Evaluation of SHOX2 and RASSF1A Methylation for the Aid in Diagnosis of Lung Cancer Based on the Analysis of FFPE Specimen

Front Oncol. 2020 Dec 14:10:565780. doi: 10.3389/fonc.2020.565780. eCollection 2020.

Authors

Juanhong Shi¹, Xue Chen¹, Long Zhang¹, Xia Fang², Yuting Liu¹, Xuyou Zhu¹, Haoyang Zhang¹, Lichao Fan³, Jun Gu¹, Suxia Zhang¹, Bin She⁴, Hongxiu Han¹, Xianghua Yi¹

Affiliations

¹ Department of Pathology, Tongji Hospital, Tongji University, Shianghai, China.
² Department of Pulmonary and Critical Care Medicine, Dongfang Hospital Affiliated to Tongji University, Shanghai, China.
³ Department of Pulmonary and Critical Care Medicine, Shanghai Pulmonary Hospital Affiliated to Tongji University, Shanghai, China.
⁴ Academic Development, Tellgen Corporation, Shanghai, China.

Abstract

Emerging molecular diagnostic methods are more sensitive and objective, which can overcome the intrinsic failings of morphological diagnosis. Here, a RT-PCR-based in vitro diagnostic test kit (LungMe^®) was developed and characterized to simultaneously quantify the DNA methylation of SHOX2 and RASSF1A in FFPE tissue specimens. The clinical manifestations were evaluated in 251 FFPE samples with specificity and sensitivity of 90.4 and 89.8%, respectively. Furthermore, the quantitative analysis shows that the degree of SHOX2 methylation was correlated with the stages of lung cancer, but not in the case of RASSF1A. Our observation indicated that the DNA methylation of SHOX2 and RASSF1A may play different roles in cancer development. Comparison of the methylation levels of SHOX2 and RASSF1A between cancer and cancer-adjacent specimens (n = 30), showed they have "epigenetic field defect". As additional clinical validation, the hypermethylation of SHOX2 and RASSF1A was detected not only in surgical operative specimens, but also in histopathological negative puncture biopsies. SHOX2 and RASSF1A methylation detection can be used to increase sensitivity and NPV, which provide us with a more accurate method of differential diagnosis and are likely to be rapidly applied in clinical examinations.

Keywords: DNA methylation; RASSF1A; SHOX2; epigenetic field defect; lung cancer.