探討新穎長鏈非編碼核糖核酸LINC02470在膀胱癌細胞中誘導上皮間質轉化之分子機轉
余枃崠1、黃晟碩2 、何嘉益3 、于承平4 、于大雄5
1國防醫學院醫學系，2國防醫學院病理及寄生蟲研究所，3國防醫學院生命科學研究所，4三軍總醫院病理部， 5三軍總醫院外科部泌尿外科
Studies on the Molecular Mechanisms of a Novel Long Intergenic Non-protein Coding RNA 02470-induced Epithelial-Mesenchymal Transition in Human Bladder Cancer Cell Lines
Ming-Fong Yee1*, Cheng-Shuo Huang2 , Jar-Yi Ho3 , Cheng-Ping Yu4 , Dah-Shyong Yu5
1School of Medicine, National Defense Medical Center, 2Graduate Institute of Pathology and Parasitology, National Defense Medical Center, 3Graduate Institute of Life Science, National Defense Medical Center, 4Department of Pathology, Tri-Service General Hospital, 5Division of Urology, Department of Surgery, Tri-Service General Hospital
Purpose: In accordance with the Ministry of Health and Welfare‘s annual report 2018, the bladder cancer-induced mortality rate in men and women is 2.2% and 1.8% respectively in Taiwan. Although the treatment of bladder cancer has improved a lot in recent years, the prognosis of patients with bladder cancer is often not as good as expected. Searching for an effective diagnostic or prognostic marker is important to achieve the goal of early diagnosis and evaluation of better treatment strategies. Long non-coding RNAs (lncRNAs) play an important role in the progression of genitourinary malignancies, including bladder cancer, renal cell carcinoma, and prostate cancer. We have found that LINC02470, a novel lncRNA, was significantly increased in bladder cancer cell lines and positively associated with aggressiveness. However, its molecular regulations and clinical meanings have not been reported. Hence, the purpose of this study is tried to elucidate the regulatory roles of LINC02470 and evaluated its potential in the diagnosis and prognosis of bladder cancers.
Materials and Methods: We selected LINC02470 as the candidate of this study upon a bioinformatic screening using lnCAR database and a comparison of its expression levels among bladder cell lines. The relative expression level of LINC02470 was determined by RTqPCR in different bladder cancer cell lines (low-grade: TSGH 8301 and TSGH 9202, and high-grade T24 and J82) and a human uroepithelial cell line (SV-HUC-1). In these cell lines, loss-of-function experiments were performed to investigate the biological effects of LINC02470 on bladder cancer cell proliferation (MTT assay), migration (Wound Healing Assay) and invasion activity (Transwell). The molecular mechanisms underlying the functions of LINC02470 were determined by western blot and RT-qPCR.
Results: LINC02470 was significantly up-regulated in high grade bladder cancers and positively associated with its aggressiveness. Further experiments demonstrated that knockdown of LINC02470 inhibited cell proliferation, migration and invasion in T24 and J82 tumor cell. In molecular regulation, The silence of LINC02470 promoted EMT of T24 and J82 bladder cancer cells by inhibiting the interaction between NOTCH4 signaling pathways and downregulating SLUG/TWIST/ZEB2 transcription factors in T24 and J82 bladder cancer cells.
Conclusion: This is the first study that demonstrates LINC02470 playing a significant regulatory role in promotion of EMT and progression of bladder cancer. LINC02470 is an encouraging novel biomarker that can be used for the clinical prognosis prediction of bladder cancer.