Heat injury-induced tRNA fragment tRF-Ile-AAT-1 inhibits proliferation and invasion of bladder urothelial cancer cells

Li JIANG; Junyong ZHANG; Peng TAN; Yang GOU; Zhiya HU; Yanjun LIN; Qing JIANG

doi:10.16016/j.1000-5404.202101122

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Heat injury-induced tRNA fragment tRF-Ile-AAT-1 inhibits proliferation and invasion of bladder urothelial cancer cells

Basic Medicine | 更新时间：2026-01-09

- Heat injury-induced tRNA fragment tRF-Ile-AAT-1 inhibits proliferation and invasion of bladder urothelial cancer cells
- Journal of Third Military Medical University Vol. 43, Issue 15, Pages: 1493-1500(2021)
- 作者机构：
  
  400010 重庆, 重庆医科大学附属第二医院泌尿外科
- 作者简介：
- 基金信息：
  
  the National Natural Science Foundation for Young Scholars of China(81803057);the "Kuanren Talents" Project of the Second Affiliated Hospital of Chongqing Medical University(KY2019Y026)
- DOI：10.16016/j.1000-5404.202101122
  CLC： R394.3;R73-37;R737.14
- Received：14 January 2021，
  
  Revised：2021-04-30，
  
  Published：15 August 2021
- 稿件说明：
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Li JIANG, Junyong ZHANG, Peng TAN, et al. Heat injury-induced tRNA fragment tRF-Ile-AAT-1 inhibits proliferation and invasion of bladder urothelial cancer cells[J]. Journal of Third Military Medical University, 2021, 43(15): 1493-1500.
DOI：

Li JIANG, Junyong ZHANG, Peng TAN, et al. Heat injury-induced tRNA fragment tRF-Ile-AAT-1 inhibits proliferation and invasion of bladder urothelial cancer cells[J]. Journal of Third Military Medical University, 2021, 43(15): 1493-1500. DOI： 10.16016/j.1000-5404.202101122.

摘要

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，

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。CCK-8结果显示T24

Heat

+Mimic组BUC细胞的增殖受到明显抑制(

0.05)；流式细胞术结果显示T24

Heat

+Mimic组的凋亡率较T24

Heat

组显著上升；Transwell实验提示T24

Heat

+Mimic组迁移的T24数量较T24

Heat

组显著下降，侵袭的T24数量较T24

Heat

组显著下降。5637

Heat

+ Inhibitor组BUC细胞增殖明显增强(

0.05)，其凋亡率较5637

Heat

组显著下降，迁移细胞数较5637

Heat

组显著上升，侵袭细胞数较5637

Heat

组显著上升。

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。

Abstract

Objective

To investigate the effect of tRNA fragment differentially expressed between high and low malignant bladder urothelial carcinoma (BUC) cells induced by heat injury on the proliferation and invasion of BUC cells.

Methods

High-grade BUC cell line T24 and low-grade BUC cell line 5637 were inflicted to heat injury. Then transfer RNA stress-induced fragments (tRFs) in these cells were sequenced before and after injury

and the tRFs whose expression decreased in T24 cells while increased in 5637 cells after heat injury were screened out

and further verified in clinical specimen by RT-PCR. The single-chain mimic and antisense strand (inhibitor) of this target tRF were constructed and transfected into heat injured T24 and 5637 cells respectively

and the cells were assigned into heat injury

vector control

mimic and inhibitor groups. Finally

CCK-8 assay

flow cytometry and Transwell test were conducted respectively to detect the regulatory effect of this tRF on the proliferation

apoptosis

migration and invasion of BUC cells.

Results

The originally selected tRFs were identified as tRF-chrM.Ser-GCT and tRF-Ile-AAT-1 through sequencing and RT-PCR in cell lines. However

when comparing the specimens from the second surgery of transurethral resection of bladder tumor (TURBT) with those from the first one

the results discovered that only tRF-Ile-AAT-1 levels were increased in low-grade BUC patients (1.61±0.16 times

0.01) while decreased in high-grade BUC patients (0.75±0.14 times

0.05). In the T24 cells experiments

when compared with the T24

Heat

group

the proliferation of BUC cells in the T24

Heat

+Mimic group was inhibited (

0.05)

the apoptotic rate was significantly improved (5.88±1.01 times

0.01)

and the number of migrated and invaded cells were both greatly diminished (0.40±0.05 times

0.01; 0.23±0.04 times

0.01). Moreover

the proliferation of BUC cells in the 5637

Heat

+ Inhibitor group was enhanced (

0.05)

the apoptotic rate was declined (0.11±0.02 times

0.01)

and the number of migrated cells as well as invaded cells were both elevated (1.81±0.29 times

0.01; 2.70±0.15 times

0.01)

as compared with the 5637

Heat

group.

Conclusion

tRNA fragment tRF-Ile-AAT-1 induced by heat injury can inhibit the proliferation and invasion in BUC cells.

关键词

Keywords

references

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