Effect of trim28 (ser473) phosphorylation on viability of hepatocellular carcinoma cells during DNA damage

中华实验外科杂志 Pub Date : 2019-12-08 DOI:10.3760/CMA.J.ISSN.1001-9030.2019.12.018

Yongliang Wang, S. Zhai, Bailin Wang, Wei Yuan, Pengzhen Wang, Xu Zhang, Weilu Jia, Hui Yuan

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Abstract

Objective To observe effect of Trim28 (ser473) phosphorylation on viability of hepatocellular carcinoma (HCC) cells during DNA damage. Methods To divide Hepg2 cells into 2 groups as SB203580+ UV group and UV group. Hepg2 cells were modeled for DNA damage by UV (Ultraviolet) irradiation. SB203580+ UV group was treated with Trim28 phosphorylation inhibitor SB203580 after UV irradiation. Expression of Trim28 and phosphorylation-Trim28 ser473 (P-Trim28 ser473)were detected by westernblot experiment. The viability of Hepg2 cells were observed by methyl thiazol tetrazolium (MTT) experiment. Results Compared with UV group (0.94±0.13), the expression value of Trim28 in SB203580+ UV group had no significant difference (t=-0.190, P>0.05). Value of phosphorylation-Trim28 in SB203580+ UV group (0.32±0.04) was obviously lower than UV group (0.72±0.05), the difference had statistical meaning (t=8.500, P<0.05); The 24, 48 and 72 hours after UV irradiation, the cell survive rate of UV group [(95±7)%, (62±10)%, (35±6)%] is higher than SB203580+ UV group [(85±7)%, (37±4)%, (20±3)%]. The difference has statistical meaning (t=-2.326, -5.212, -4.577, P<0.05). Conclusion The viability of HCC cells during DNA damaged can be reduced by inhibition for Trim28 phosphorylation. Key words: Trim28; Phosphorylation; Cell survival

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trim28（ser473）磷酸化对肝细胞癌细胞DNA损伤生存能力的影响

目的观察Trim28（ser473）磷酸化对肝细胞癌（HCC）细胞DNA损伤后生存能力的影响。方法将Hepg2细胞分为SB203580+UV组和UV组。通过UV（紫外线）照射对Hepg2细胞的DNA损伤进行建模。SB203580+UV组在UV照射后用Trim28磷酸化抑制剂SB203580处理。Western印迹实验检测Trim28和磷酸化-Trim28-ser473（P-Trim28-ser473）的表达。MTT法观察Hepg2细胞的活力。结果与UV组比较（0.94±0.13），SB203580+UV组Trim28的表达值无显著性差异（t=-0.190，P>0.05），磷酸化-Trim28在SB203580+TUV组的表达值（0.32±0.04）明显低于UV组（0.72±0.05），差异有统计学意义（t=8.500，P<0.05）；紫外线照射后24、48和72小时，紫外线组的细胞存活率[（95±7）%、（62±10）%、[（35±6）%]高于SB203580+紫外线组[（85±7）%、[（37±4）%、]。差异有统计学意义（t=-2.326，-5.212，-4.577，P<0.05）。关键词：Trim28；磷酸化；细胞存活率

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中华实验外科杂志

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