OncoTargets and Therapy (Oct 2020)
Knockdown of LncRNA DLEU2 Inhibits Cervical Cancer Progression via Targeting miR-128-3p
Abstract
Bofei Wang,1 Jing Hang,2– 4 Weiling Li,5 Wanqiong Yuan6,7 1Department of Obstetrics and Gynecology, Weifang NO.2 People’s Hospital; 2Center for Reproductive Medicine, Department of Obstetrics and Gynecology, Beijing, People’s Republic of China; 3Peking University Third Hospital, Key Laboratory of Assisted Reproduction, Ministry of Education, Beijing, People’s Republic of China; 4Beijing Key Laboratory of Reproductive Endocrinology and Assisted Reproduction, Beijing, People’s Republic of China; 5Department of Obstetrics and Gynecology, Affiliated Yixing Hospital of Jiangsu University, Jiangsu, People’s Republic of China; 6Department of Orthopedics, Peking University Third Hospital, Beijing, People’s Republic of China; 7Beijing Key Laboratory of Spinal Disease, Beijing, People’s Republic of ChinaCorrespondence: Weiling LiDepartment of Obstetrics and Gynecology, Affiliated Yixing Hospital of Jiangsu University, No. 75. Tongzhenguan Road, Yixing, People’s Republic of ChinaTel +86-13921327907Email [email protected] YuanDepartment of Orthopedics Peking University Third Hospital, 49 North Garden Road, Haidian District, Beijing, People’s Republic of ChinaTel +86-15120092324Fax +86 10-82266611Email [email protected]: Cervical cancer is one of the most common female malignancies worldwide and represents a major global health challenge. The fast growth of tumor and high rates of metastasis still lead to a poor prognosis of cervical cancer patients. It is urgent to clarify the mechanism and identify predictive biomarkers for the treatment of cervical cancer. Long non-coding RNAs (LncRNAs) have been identified in cervical cancer and are related to malignant phenotypes of cervical cancer cells. However, the roles and mechanism of LncRNA deleted in lymphocytic leukemia (DLEU2) in the tumorigenesis and progression of cervical cancer remain unknown.Materials and Methods: qPCR was performed to analyze the expression of DLEU2, Cyclin D1, CDK4, Bax, Bcl2 and mi-128-3p. Western blot was performed to detect the cell cycle hallmarks expression. CCK8 was used to examine cell proliferation. Cellular apoptosis was analyzed by Hoechst 33,258 staining and AV/PI staining with flow cytometry. Cell cycle was analyzed by flow cytometry. The xenograft model in nude mice was used to elucidate the function of DLEU2 in vivo. Bioinformatics analysis and luciferase reporter assay were proceeded to clarify whether miR-128-3p directly binds with lncRNA DLEU2. Pull‑down assay and RNA-binding protein immunoprecipitation assay were used for exploring the relationship between DLEU2 and miR-128-3p.Results: We demonstrated that DLEU2 was upregulated in cervical cancer tumor tissues. Downregulation of DLEU2 inhibited cell proliferation, induced apoptosis and cell cycle arrest at G2/M phase of cervical cancer cells in vitro, and suppressed tumor growth in vivo. Further, LncRNA DLEU2 is one of the targets of miR-128-3p. miR-128-3p inhibitor abrogated the cell proliferation suppressed by knockdown of DLEU2, apoptosis induced by knockdown of DLEU2 and reversed the expression of cell cycle hallmarks regulated by knockdown of DLEU2.Conclusion: Taken together, these results suggested knockdown of DLEU2 inhibited cervical cancer progression via targeting miR-128-3p.Keywords: cervical cancer, LncRNAs, DLEU2, miR-128-3p, tumor growth
