| Ye Tianxia,Ying Yimeng,Ma Shumei,Liu Xiaodong.Mechanism of lncRNA PRMT5-AS1 regulating radiation-induced ferroptosis in hepatocellular carcinoma cells[J].Chinese Journal of Radiological Medicine and Protection,2023,43(12):954-961 |
| Mechanism of lncRNA PRMT5-AS1 regulating radiation-induced ferroptosis in hepatocellular carcinoma cells |
| Received:September 24, 2023 |
| DOI：10.3760/cma.j.cn112271-20230924-00098 |
| KeyWords:PRMT5-AS1 Ion irradiation Cell death Liver cancer Ferroptosis |
| FundProject:国家自然科学基金(81972969) |
| Author Name | Affiliation | E-mail | | Ye Tianxia | School of Public Health and Management, Wenzhou Medical University, Wenzhou 325035, China | | | Ying Yimeng | School of Public Health and Management, Wenzhou Medical University, Wenzhou 325035, China | | | Ma Shumei | School of Public Health and Management, Wenzhou Medical University, Wenzhou 325035, China | | | Liu Xiaodong | School of Public Health and Management, Wenzhou Medical University, Wenzhou 325035, China | liuxd2014@126.com |
|
| Hits: 659 |
| Download times: 236 |
| Abstract:: |
| Objective To evaluate the effect of long non-coding RNA PRMT5-AS1 on ferroptosis of hepatocellular carcinoma cell(HCC) after ionizing irradiation.Methods The PRMT5-AS1 overexpression model was constructed in MHCC-97H cells and the PRMT5-AS1 knockdown model was constructed in HepG2 cells. X-ray irradiation(IR) was performed with an absorbed dose of 10 Gy and a dose rate of 3 Gy/min. Western blot and qRT-PCR were used to detect gene expression. The effect of PRMT5-AS1 expression on lipid peroxidation and ferroptosis of HCC after IR was detected by Trypan blue staining flow cytometry. The effect of PRMT5-AS1 expression on the death of HCC after IR was detected by CCK-8 assay. Dual luciferase assay to detect the binding of let-7c-5p to PRMT5-AS1 and SLC7A11.Results Overexpression of PRMT5-AS1 in MHCC-97H cells could significantly reduce cell death induced by IR (Vector vs. PRMT5-AS1:27.57% vs.18.30%,t=14.94,P<0.05). Knockdown of PRMT5-AS1 in HepG2 cells significantly increased cell death induced by IR (siNC vs. siPRMT5-AS1: 17.26% vs. 28.26%, t=13.63, P<0.05). Flow cytometry result show that overexpression of PRMT5-AS1 can significantly inhibit the increase of intracellular lipid ROS level induced by IR (Vector vs. PRMT5-AS1: 17.01% vs. 12.52%, t=12.80, P<0.05), and knockdown of PRMT5-AS1 significantly increases the lipid ROS level induced by IR (siNC vs. siPRMT5-AS1: 14.54% vs. 17.72%, t=5.93, P<0.05). The result of CCK-8 experiment showed that overexpression of PRMT5-AS1 could significantly inhibit Erastin induced cell activity reduction (Vector vs. PRMT5-AS1:87.92% vs. 109.06%, t=2.87, P<0.05), and knockdown of PRMT5-AS1 could promote Erastin's inhibitory effect on cell activity (siNC vs. siPRMT5-AS1: 82.56% vs. 60.58%, t=38.35, P<0.05). Western blot and fluorescent quantitative PCR result showed that the protein and mRNA levels of SLC7A11 were significantly increased after overexpression of PRMT5-AS1 (t=26.24, P<0.05), and the protein and mRNA levels of SLC7A11 were significantly decreased after knockdown of PRMT5-AS1 (t=5.60, P<0.05). The correlation between PRMT5-AS1 and let-7c-5p was confirmed by luciferase report gene experiment (t=9.74, P<0.05). The result of luciferase reporter gene experiment showed that PRMT5-AS1 could form ceRNA network with let-7c-5p to regulate SLC7A11. Let-7c-5p was able to reverse the increase in SLC7A11 expression levels, decrease in Lipid-ROS levels and cell death induced by overexpression of PRMT5-AS1 (t=3.01, 4.11, P<0.05). And knockdown of SLC7A11 reversed Lipid-ROS inhibition and reduced cell death caused by PRMT5-AS1(t=21.35, 7.15, P<0.05). Conclusions LncRNA PRMT5-AS1 inhibits IR-induced ferroptosis in HCC through the PRMT5-AS1/let-7c-5p/SLC7A11 axis. |
| HTML View Full Text View/Add Comment Download reader |
| Close |
|
|
|
