敲减USP25对乳腺癌细胞生物学行为的影响

Effect of knocking down USP25 on biological behavior of breast cancer cells

目的:探讨乳腺癌细胞中敲减泛素特异性蛋白酶25(USP25)后细胞生物学行为的改变。方法:采用慢病毒载体构建6条shRNA-USP25干扰序列(LV-shRNA-USP25-1~LV-shRNA-USP25-6),分别转染乳腺癌MCF-7细胞以干扰USP25的表达,并检测敲减效率,筛选出最佳干扰序列用于实验。采用CCK8试剂盒法、Transwell侵袭实验、流式细胞术检测敲减USP25后乳腺癌细胞增殖、侵袭能力及凋亡的变化。结果:KD1组(转染LV-shRNA-USP25-1组)为干扰效率最高的序列,USP25蛋白和mRNA的敲减率分别为57.8%、58.3%(P<0.001);敲减USP25后乳腺癌MCF-7细胞增殖、侵袭能力受到抑制,且出现明显凋亡(P<0.001)。结论:USP25可能与乳腺癌细胞的增殖、侵袭及凋亡有密切关系,其可能在乳腺癌的恶性进展中扮演着重要角色。

Objective:To investigate the changes of biological behavior after knocking down ubiquitin specific protease 25(USP25)in breast cancer cells.Methods:6 shRNA-USP25 interference sequences were constructed by lentiviral vector(LV-shRNA-USP25-1~LV-shRNA-USP25-6),and transfected into MCF-7 breast cancer cells to interfere with the expression of USP25.The knockdown efficiency was detected and the best interference sequence was screened for the experiment.CCK8 kit,Transwell test and flow cytometry were used to detect the proliferation,invasion and apoptosis of breast cancer cells after knocking down USP25.Results:Group KD1(transfected with LV-shRNA-USP25-1)was the most effective sequence,with the knockdown rates of USP25 protein and mRNA of 57.8%and 58.3%,respectively(P<0.001).After USP25 was knocked down,the proliferation and invasion of MCF-7 breast cancer cells were inhibited,and showed significant apoptosis(P<0.001).Conclusion:USP25 may be closely related to the proliferation,invasion and apoptosis of breast cancer cells,and may play an important role in the malignant progression of breast cancer.