| Han Yang,Yu Jingping,Sun Zhiqiang,et al.TUT4 affects the radiosensitivity of esophageal epithelial cells by regulating the uridylation of miR132/212 clusters[J].Chinese Journal of Radiological Medicine and Protection,2021,41(11):801-807 |
| TUT4 affects the radiosensitivity of esophageal epithelial cells by regulating the uridylation of miR132/212 clusters |
| Received:May 18, 2021 |
| DOI：10.3760/cma.j.issn.0254-5098.2021.11.001 |
| KeyWords:Radiation esophagitis TUT4 miR132/212 Uridylation |
| FundProject:国家自然科学基金（82073339；81773224）；江苏省自然科学基金（BK20191157） |
| Author Name | Affiliation | E-mail | | Han Yang | Department of Radiotherapy, Changzhou No.2 People's Hospital, Affiliated Hospital of Nanjing Medical University 213003, Changzhou China | | | Yu Jingping | Department of Radiotherapy, Changzhou No.2 People's Hospital, Affiliated Hospital of Nanjing Medical University 213003, Changzhou China | | | Sun Zhiqiang | Department of Radiotherapy, Changzhou No.2 People's Hospital, Affiliated Hospital of Nanjing Medical University 213003, Changzhou China | | | Luo Judong | Department of Radiotherapy, Changzhou No.2 People's Hospital, Affiliated Hospital of Nanjing Medical University 213003, Changzhou China | judongluo@163.com |
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| Abstract:: |
| Objective To validate the effect of TUT4 on the radiosensitivity of esophageal epithelial cells (HEEC) by regulating the uridylation of miR132/212 clusters. Methods The expression of TUT4 in HEEC at 0, 6, 18, 24 and 48 h after 0, 2, 4, 6 and 8 Gy X-ray irradiation was detected by PCR. The HEEC cells were divided into four groups:NC group, shTUT4 group, 6 Gy group, and 6 Gy+shTUT4 group. The effects of TUT4 on cell radiosensitivity, cell proliferation, cell cycle, mitochondrial damage, and oxygen free radical production were detected respectively. The effect of down-regulated TUT4 expression on miR132/212 uridylation was detected by RT-PCR, and the radiosensitivity of HEEC with overexpression of miR132/212 or miR132/212+UUU was detected by clone formation and proliferation assay, respectively. Proliferation assay was performed to detect the proliferation of HEEC when TUT4 expression was down-regulated and miR132/212 or miR132/212+UUU was overexpressed. Results TUT4 expression increased after different doses of X-ray irradiation (t=12.84, 62.06, 27.86, 32.43, P<0.05). Downregulation of TUT4 expression increased the radiosensitivity of HEEC (t=13.2, 5.85, 7.31, P<0.05) with a SERD0 of 1.41 and D0=0.79, Dq=1.61, SF2=0.47. Compared with 6 Gy group, cell proliferation in 6 Gy+ shTUT4 group was decreased (t=7.12, 13.63, P<0.05), cells in S phase were increased (t=11.98, P<0.05), mitochondrial damage was increased (t=11.98, P<0.05), and ROS level was increased (t=15.65, P<0.05). Down-regulation of TUT4 expression increased miR132/212 expression and decreased miR132/212+UUU expression (t=27.90, 60.99, P<0.05). Overexpression of miR132/212 increased the radiosensitivity of HEEC, and overexpression of miR132/212+UUU decreased the radiosensitivity of HEEC, with SERD0 of 1.20 and 0.71, respectively. Cell proliferation of shTUT4 +miR132/212 group waslower than that of shTUT4 group(t=4.76, 7.65, P<0.05), and cell proliferation of shTUT4 +miR132/212+UUU group was higher than that of shTUT4 (t=7.22, P<0.05). Conclusions X-ray irradiation increased the expression of TUT4 in HEEC, and the down-regulation of TUT4 reduced HEEC radiosensitivity and radiation damage, where the uridylation of miR132/212 was involved in. |
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