DataSheet_1_tRNA-Derived Fragment tRF-Glu-TTC-027 Regulates the Progression of Gastric Carcinoma via MAPK Signaling Pathway.docx

Transfer RNA-derived RNA fragments (tRFs) belong to non-coding RNAs (ncRNAs) discovered in most carcinomas. Although some articles have demonstrated the characteristics of tRFs in gastric carcinoma (GC), the underlying mechanisms still need to be elucidated. Meanwhile, it was reported that the MAPK pathway was momentous in GC progression. Thus we focused on investigating whether tRF-Glu-TTC-027 could act as a key role in the progression of GC with the regulation of the MAPK pathway. We collected the data of the tRNA-derived fragments expression profile from six paired clinical GC tissues and corresponding adjacent normal samples in this study. Then we screened tRF-Glu-TTC-027 for analysis by using RT-PCR. We transfected GC cell lines with tRF-Glu-TTC-027 mimics or mimics control. Then the proliferation, migration, and invasion assays were performed to assess the influence of tRF-Glu-TTC-027 on GC cell lines. Fluorescence in situ hybridization assay was conducted to confirm the cell distribution of tRF-Glu-TTC-027. We confirmed the mechanism that tRF-Glu-TTC-027 influenced the MAPK signaling pathway and observed a strong downregulation of tRF-Glu-TTC-027 in clinical GC samples. Overexpression of tRF-Glu-TTC-027 suppressed the malignant activities of GC in vitro and in vivo. MAPK signaling pathway was confirmed to be a target pathway of tRF-Glu-TTC-027 in GC by western blot. This is the first study to show that tRF-Glu-TTC-027 was a new tumor-suppressor and could be a potential object for molecular targeted therapy in GC.

转运RNA衍生RNA片段（transfer RNA-derived RNA fragments, tRFs）属于非编码RNA（non-coding RNAs, ncRNAs）家族，在多数恶性肿瘤中均有检出。尽管已有研究揭示了胃癌（gastric carcinoma, GC）中tRFs的表达特征，但其具体作用机制仍有待阐明。同时，已有研究证实丝裂原活化蛋白激酶（mitogen-activated protein kinase, MAPK）通路在胃癌进展中发挥关键调控作用。因此本研究聚焦于探究tRF-Glu-TTC-027是否通过调控MAPK通路，在胃癌进展中扮演关键角色。本研究从6例配对的临床胃癌组织及对应癌旁正常组织中获取了tRNA衍生片段的表达谱数据。随后通过逆转录聚合酶链反应（reverse transcription polymerase chain reaction, RT-PCR）筛选出tRF-Glu-TTC-027用于后续实验分析。将tRF-Glu-TTC-027模拟物（mimics）及对照模拟物转染至胃癌细胞系中，随后通过细胞增殖、迁移及侵袭实验，评估tRF-Glu-TTC-027对胃癌细胞系的生物学影响。采用荧光原位杂交（fluorescence in situ hybridization, FISH）实验验证tRF-Glu-TTC-027在胃癌细胞内的分布定位。本研究明确了tRF-Glu-TTC-027调控MAPK信号通路的具体机制，并观察到临床胃癌样本中tRF-Glu-TTC-027呈现显著低表达。过表达tRF-Glu-TTC-027可在体内外抑制胃癌细胞的恶性生物学行为。经蛋白质印迹（western blot, WB）实验验证，MAPK信号通路是tRF-Glu-TTC-027在胃癌中的靶调控通路。本研究首次证实tRF-Glu-TTC-027是一种新型抑癌非编码RNA，有望成为胃癌分子靶向治疗的潜在靶点。