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Chinese Journal of Cell and Stem Cell(Electronic Edition) ›› 2021, Vol. 11 ›› Issue (06): 337-342. doi: 10.3877/cma.j.issn.2095-1221.2021.06.003

• Original Research • Previous Articles     Next Articles

TET3 promotes proliferation of renal clear cell carcinoma cells

huiyue Dong1, xiao Zhang1, ling Zhu1, yi Zhang1, jingjing Sun1, jun Lu1,()   

  1. 1. Laboratory of Basic Medicine, Fujian Provincial Key Laboratory of Transplant Biology, 900th Hospital of Joint Logistics Support Force, Fuzhou 350025, China
  • Received:2020-11-17 Online:2021-12-01 Published:2022-01-05
  • Contact: jun Lu

Abstract:

Objective

To analyze TET3 expression in renal clear cell carcinoma, verify the role of TET3 in renal clear cell carcinoma cells and explore its mechanism.

Methods

The expression of TET3 gene in normal tissues and their corresponding tumor tissues was analyzed by Fire Browse. The expression of TET3 in renal clear cell carcinoma and normal tissues was analyzed by TCGA database. Human renal carcinoma cells (ACHN) cells were stably transfected with shRNA-NC, shRNA-TET3. Cell proliferation was detected on 24, 48 and 72 hours by the Cell Counting Kit-8 (CCK-8) assay. The number of clones in transfected cell was counted by clone formation experiment. The migration of transfected cells was detected by transwell co-culture migration test. The expression of protein TET3, PARP, p-P38, P38, p-ERK, and ERK was determined by Western blot. Independent sample t test was used for comparison between two groups. Dunnett's-t test was used to compare multiple experimental groups with the same control group.

Results

Based on the data excavation, the expression of TET3 inrenal clear cell carcinoma was higher than that in the normal tissues. Compared with transfected shRNA-NC, the absorbance of ACHN cells after shRNA-TET3 transfection was not significantly different at 24 h (0.2501±0.0065 vs 0.2500±0.0073) and 48 h (0.3964±0.01402 vs 0.3711±0.011) , the difference was not statistically significant (P > 0.05) ,the absorbance value at 72 h (0.4303±0.0287 vs 0.3641±0.0230) is significantly reduced, the difference is statistically significant (P < 0.05) . Compared with transfection of shRNA-NC, the number of cell clones formed after transfection of shRNA-TET3 [ (60.00±6.00) vs (37.00±9.00) ] was significantly reduced, the difference is statistically significant (P < 0.01) . Compared with transfected shRNA-NC, there is no significant difference in the number of migrating cells after transfection of shRNA-TET3 [(49.00±4.00) vs (50.00±3.00) ], the difference was not statistically significant (P > 0.05) . Compared with transfected shRNA-NC, there is no significant difference in PARP, ERK, and P38 protein molecules after transfection of shRNA-TET3, the expression of p-ERK and p-P38 protein molecules is significantly reduced.

Conclusion

TET3 is highly expressed in renal clear cell carcinoma, Knockdown of TET3 can inhibit the expression of p-ERK and p-P38 protein to restrain tumor cell proliferation in vitro. These results provide clues and basis for TET3 as a marker for diagnosis of renal clear cell carcinoma.

Key words: Renal clear cell carcinoma, TET3, Data excavation, ACHN, Cell proliferation

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