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OBJECTIVE To investigate the inhibitory effect of berberine (BER) on the invasion and migration of human renal carcinoma cells and its potential mechanism. METHODS Using human renal carcinoma OSRC-2 cell as object, alamarBlue assay was adopted to detect the inhibitory effects of 0 (control group), 25, 50, 75, 100, 125, 150, 175 and 200 μmol/L BER on the proliferation of OSRC-2 cell after treatment for 24 h and 48 h. After treated with 0(control group), 50, 100 μmol/L BER for 48 h, the effect of BER on cell cycle was analyzed by flow cytometry. The migration of OSRC-2 cells in 24 h and 36 h was observed by cell scratch test, and the invasion ability of OSRC-2 cells in 24 h was detected by Transwell assay. The protein expression of methyltransferase-like 3 (METTL3) was detected by Western blot after treatment for 48 h, and RNA methylation quantification kit was used to detect the levels of N6-methyladenosine (m6A) in OSRC-2 cells. RESULTS Compared with control group, BER at different concentrations could significantly decrease the survival rate of OSRC-2 cells (P<0.01), and showed a dose-dependent and time-dependent manner. After 48 h of BER treatment at 50, 100 μmol/L, the cell was arrested in G0/G1 phase (P<0.01). Compared with control group, the migration and invasion abilities of cells in 50, 100 μmol/L BER group were significantly decreased (P<0.05 or P<0.01); the protein expression of METTL3 and the level of m6A in RNA were significantly decreased (P<0.01). CONCLUSIONS BER can inhibit level of m6A by down-regulating the expression of METTL3, thereby inhibiting the invasion and metastasis of human renal carcinoma cells.
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Objective:To investigate the effects of ultra-high dose rate radiation (FLASH-RT) and conventional radiation (CONV-RT) on plasma metabolites in glioma mice.Methods:Tocally 21 male C57BL/6J mice bearing intracranial glioma xenograft were randomly divided into healthy control group ( n=3), CONV-RT group ( n=9) and FLASH-RT group ( n=9). The CONV-RT group was administered a single dose of 24 Gy radiation on the head of mice at a dose rate of 0.4 Gy/s, and the FLASH-RT group was administered a single dose of 24 Gy radiation on the head of mice at a dose rate of 60 Gy/s, and the healthy control group was given 0 Gy pseudoradiation under the same condition. Mice blood was collected through the inner canthus vein for plasma separation at 1, 3 and 7 d after radiation in the two radiation groups, and the blood plasma of healthy control group was collected at 7 days after sham radiation. The changes in plasma metabolites were detected by the non-targeted metabolomics based on liquid chromatography mass spectrometry tandem platform. Results:After irradiation, the metabolites in plasma of two irradiation groups had significant difference. Compared with the healthy control group, 12 and 5 differential metabolites were screened out in the FLASH-RT group and CONV-RT group at three time points, respectively. The difference of plasma metabolites had the largest value at 1 day and decreased at 3 and 7 d after radiation. The arachidonic acid and isovaleric acid, involving arachidonic acid metabolism, biosynthesis of unsaturated fatty acids, and tyrosine metabolism pathways were screened in both FLASH-RT group and CONV-RT group, and the 10 differential metabolites, mainly involving energy metabolism and redox reactions, only existed in the FLASH-RT group.Conclusions:Arachidonic acid and isovaleric acid may be the common sensitive biomarkers to different radiation patterns, which provides ideas for further exploring the molecular mechanism of FLASH-RT in the treatment of glioma.