NUP88基因变化对乳腺癌细胞系BT-20增殖侵袭生物学行为的影响

Effect of NUP88 gene on proliferation and invasion biological behavior of breast cancer cell line BT-20

目的:探讨NUP88基因表达量升高或降低对乳腺癌细胞系BT-20细胞增殖能力、凋亡能力与侵袭能力的影响。方法:构建NUP88重组腺病毒表达载体以及NUP88 RNA干扰腺病毒载体,分别转染乳腺癌BT-20细胞获得NUP88过表达BT-20细胞以及NUP88低表达BT-20细胞并检测NUP88 mRNA和蛋白表达情况,随后通过CCK-8检测各组BT-20细胞增殖能力,通过流式双染检测各组BT-20细胞凋亡情况及通过Transwell侵袭实验检测各组BT-20细胞侵袭能力,并通过Western blot检测凋亡和侵袭相关蛋白表达变化。结果:成功获得NUP88 mRNA及蛋白高表达和低表达BT-20细胞;NUP88基因过表达导致细胞增殖能力和侵袭细胞数量显著高于正常BT-20细胞水平,而凋亡率则降低(P<0.05);NUP88基因低表达导致细胞增殖能力和侵袭细胞数量显著低于正常BT-20细胞水平,而凋亡率则升高(P<0.05);NUP88基因过表达导致抗凋亡蛋白Bcl-2和黏附蛋白β-catenin水平显著高于正常BT-20细胞水平,而促凋亡蛋白Bax和黏附蛋白E-cadherin显著低于正常BT-20细胞水平(P<0.05);NUP88基因低表达导致Bcl-2和β-catenin水平显著低于正常BT-20细胞水平,而Bax和E-cadherin显著高于正常BT-20细胞水平(P<0.05)。结论:NUP88基因通过调控凋亡相关蛋白Bax与Bcl-2和黏附蛋白E-cadherin与β-catenin水平调控BT-20细胞的增殖、凋亡和侵袭能力。

Objective To observe the effect of low-expression or over-expression of NUP88 gene on the proliferation and invasion ability of breast cancer cell line BT-20. Methods： NUP88 recombinant adenovirus expression vector and NUP88 RNAi adenovirus vector were transfected into breast cancer BT-20 cells to obtain BT-20 cells over-expressing NUP88 and BT-20 cells lower- expressing NUP88 and then detected the expression of NUP88 mRNA and NUP88 protein. After that,the apoptosis of BT-20 cells was detected by flow cytometry and the invasion and metastasis of BT-20 cells were detected by Transwell invasion assay. The expression of apoptosis protein and invasion and metastasis proteins were detected by Western blot. Results： BT-20 cell with the over expression levels of NUP88 mRNA and NUP88 protein and BT-20 cell with the low expression levels of NUP88 mRNA and NUP88 protein were structured. The over-expression of NUP88 gene led to proliferation rate and the number of invasive cells were significantly higher than BT-20 cells,apoptosis cells were significantly lower than BT-20 cells（P〈0. 05）. However,the low-expression of NUP88 gene led to pro-liferation rate and the number of invasive cells were significantly lower than BT-20 cells, apoptosis cells was significantly higher than BT-20 cells（P〈0. 05）. The over-expression of NUP88 gene led to Bcl-2 and 茁 -catenin level were significantly higher than that of BT-20 cells,and Bax and E-cadherin level were significantly lower than that of BT-20 cells（ P〈0. 05）. However,the low-expression of NUP88 gene led to Bcl-2 and β -catenin level were significantly lower than that of BT-20 cells, and Bax and E-cadherin level were significantly higher than that of BT-20 cells（P 〈0. 0 5 ） . Conclusion： NUP88 gene regulates the proliferation and invasion and migration ability of breast cancer cells by regulating the expression of Bax, Bcl-2, E-cadherin and β -catenin. It has an important significance in the target treatment of breast cancer.