肝癌细胞DNA损伤时三结构域蛋白28磷酸化对肝癌细胞生存能力的影响
Effect of trim28 (ser473) phosphorylation on viability of hepatocellular carcinoma cells during DNA damage
目的 观察当肝癌细胞的DNA损伤时三结构域蛋白28(Trim28)磷酸化对其生存能力的影响.方法 将Hepg2细胞系分为SB203580+ UV组及UV组.用紫外线(UV)照射Hepg2细胞建立Hepg2细胞DNA损伤的模型,SB203580+ UV组在UV照射后使用Trim28磷酸化抑制剂SB203580处理细胞.通过蛋白质印迹法(Western blot)检测上述处理后两组细胞的Trim28、磷酸化Trim28 ser473(p-Trim28 ser473)的表达;通过噻唑蓝(MTT)实验观察上述处理后Hepg2细胞的生存能力.结果 SB203580+ UV(0.96±0.16)组与UV组(0.94±0.13)的Trim28表达比较,差异无统计学意义(t=-0.190,P> 0.05);,SB203580+UV组p-Trim28(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、72 h UV组细胞生存率[(95±7)%、(62±10)%、(35±6)%]明显高于SB203580+ UV组[(85±7)%、(37±4)%、(20±3)%],差异有统计学意义(t=-2.326、-5.212、-4.577,P<0.05).结论 当肝癌细胞DNA损伤时抑制Trim28(ser473)磷酸化可以降低癌细胞的生存能力.
更多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-Trirn28 ser473 (P-Trim28 ser473) were detected by westernblot experiment.The viability of Hepg2 cells were observed by methyl thiazol tetrazolium (MTF) 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.
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