LONG NON-CODING RNA AFAP1-AS1 REGULATES EMT RELATED TRANSCRIPTION FACTORS IN THE PATHOGENESIS OF ENDOMETRIOSIS

Shumei Liu¹, Junxian Ma², Xiaoling Zhu¹, Ying Gao³, Zihan Geng⁴, Guoyan Liu^5*

¹Obstetrics and Gynecology, Sino-Singapore Eco-City Hospital of Tianjin Medical University, Tianjin, 300467, China - ²Gynaecology, The First Affiliated Hospital of Henan University of Science and Technology, Luoyang, 471000, China - ³Gynaecology, Hebei Qinhuangdao Haigang Hospital, Qinhuangdao, 066000, China - ⁴Medical Imaging, Jiangsu Nantong University Medical School, Nantong, 226007, China - ⁵Gynaecology, Tianjin Medical University General Hospital, Tianjin, 300040, China

Objective: To analyze the role of long non-coding RNA (LNC RNA) actin fiber associated protein antisense RNA1 (AFAP1-AS1) in the pathogenesis of endometriosis by regulating epithelial-mesenchymal transition (EMT) related transcription factors.

Methods: Twenty-three patients with EM treated in our hospital from July 2018 to March 2020 were randomly selected as the experimental group, and 23 patients with embryo transfer due to tubal infertility were selected as the control group. The expression levels of AFAP1-AS1 and zinc finger binding protein 1 (ZEB1) in EM tissues were measured by real-time fluorescent quantitative PCR. Primary endometrial cells (Ishikawa cells) were selected and infected with AFAP1-AS1 lentiviral solution. The negative control group was set up. The low expression group of AFAP1-AS1 and the control group of afap1-as1 were obtained. The expression level of afap1-as1 was detected by real-time fluorescent quantitative PCR. Real-time PCR and Western blot were used to detect the expression of EMT marker gene and protein in the two groups. MTT method was used to determine the cell proliferation ability of the two groups. Transwell cell migration and invasion assay was used to determine the cell migration and invasion. The expression of ZEB1 in the two groups was detected by real-time fluorescent quantitative PCR and Western blot.

Results: Compared with the control group, the expression levels of afap1-as1 and ZEB1 in the experimental group were significantly increased (P< 0.05). Compared with the control group of FAP1-AS1, the expression levels of AFAP1-AS1, N-cadherin, vimentin, and ZEB1 in the low expression group of AFAP1-AS1 were significantly decreased, and the expression levels of E-cadherin and keratin were significantly increased (P< 0.05). Compared with the fap1-as1 control group, the proliferation, invasion, and migration ability of the afap1-as1 low expression group were significantly decreased (P< 0.05).

Conclusion: Abnormal expression of LNC RNA afap1-as1 was found in ectopic tissues of EM patients. Downregulation of LncRNA AFAP1-AS1 can significantly inhibit cell proliferation, invasion, and migration, and inhibit the pathological development of EM, which may be achieved by inhibiting the expression of EMT-related transcription factors.