热损伤诱导的tRNA碎片tRF-Ile-AAT-1抑制膀胱尿路上皮癌细胞的增殖与侵袭
Journal of third military medical university(2021)
Abstract
目的 探讨热损伤诱导高度恶性与低度恶性膀胱尿路上皮癌(bladder urothelial cancer,BUC)细胞之间差异表达的tRNA碎片对BUC细胞增殖与侵袭的影响.方法 对高度恶性BUC细胞株T24和低度恶性BUC细胞株5637行热损伤处理.对热损伤前后细胞进行tRNA碎片(transfer RNA stress-induced fragments,tRFs)测序,筛选出热损伤后在T24中表达下降且在5637细胞中表达上升的tRFs,并对其行细胞株及临床标本RT-PCR检测及验证.构建目的tRF的单链模拟物(mimic)和反义链(inhibitor),并转染至热损伤后的T24及5637细胞中,分组为:相对应的Heat组、载体对照组、Mimic组、Inhibitor组.通过CCK-8、流式细胞术、Transwell实验检测tRF对BUC细胞增殖、凋亡、迁移及侵袭的调控作用.结果 通过测序及细胞株RT-PCR检测,所筛选出的tRFs分别为tRF-chrM.Ser-GCT、tRF-Ile-AAT-1,而只有tRF-Ile-AAT-1在低级别BUC患者行第2次经尿道膀胱肿瘤电切(transurethral resection of bladder tumor,TURBT)所取标本中表达量较第1次TURBT的标本显著升高[(1.61±0.16)倍,P<0.01];且在高级别BUC患者中表达量较第1次TURBT标本显著降低[(0.75±0.14)倍,P<0.05].CCK-8结果显示T24Heat+Mimic组BUC细胞的增殖受到明显抑制(P<0.05);流式细胞术结果显示T24Heat+Mimic 组的凋亡率较 T24Heat组显著上升[(5.88±1.01)倍,P<0.01];Transwell 实验提示 T24Heat+Mimic组迁移的T24数量较T24Heat组显著下降[(0.40±0.05)倍,P<0.01],侵袭的T24数量较T24Heat组显著下降[(0.23±0.04)倍,P<0.01].5637Heat+Inhibitor组BUC细胞增殖明显增强(P<0.05),其凋亡率较5637Heat组显著下降[(0.11±0.02)倍,P<0.01],迁移细胞数较5637Heat组显著上升[(1.81±0.29)倍,P<0.01],侵袭细胞数较5637Heat组显著上升[(2.70±0.15)倍,P<0.01].结论 热损伤诱导的tRNA碎片tRF-Ile-AAT-1,可抑制BUC细胞的增殖与侵袭能力.
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Key words
bladder urothelial carcinoma,transfer rna fragments,heat injury,trf-ile-aat-1
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