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
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摘要
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 proliferation rate and the number of invasive cells were significantly lower than BT-20 cells,apoptosis cells was significantly higher than BT-20cells( P<0. 05). The over-expression of NUP88 gene led to Bcl-2 and β-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. 05). 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.
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 proliferation rate and the number of invasive cells were significantly lower than BT-20 cells,apoptosis cells was significantly higher than BT-20cells( P<0. 05). The over-expression of NUP88 gene led to Bcl-2 and β-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. 05). 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.
引文
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[2]Yuan J,Liu M,Yang L,et al.Acquisition of epithelialmesenchymal transition phenotype in the tamoxifen-resistant breast cancer cell:a new role for G protein-coupled estrogen receptor in mediating tamoxifen resistance through cancer-associated fibroblastderived fibronectin andβ1[J].Breast Cancer Res,2015,17(1):1-18.
[3]Nelson HD,Fu R,Cantor A,et al.Effectiveness of breast cancer screening:systematic review and meta-analysis to update the 2009U.S.preventive services task force recommendation[J].Ann Intern Med,2016,47(4):18-21.
[4]Swain SM,Baselga J,Kim SB,et al.Pertuzumab,trastuzumab,and docetaxel in HER2-positive metastatic breast cancer[J].New Engl J Med,2015,372(8):724-734.
[5]Francis PA,Regan MM,Fleming GF,et al.Adjuvant ovarian suppression in premenopausal breast cancer[J].New Engl J Med,2015,372(5):436-446.
[6]Marinello PC,Da ST,Panis C,et al.Mechanism of metformin action in MCF-7 and MDA-MB-231 human breast cancer cells involves oxidative stress generation,DNA damage,and transforming growth factorβ1 induction[J].Tumor Biol,2016,78(4):449-454.
[7]Yi S,Wen L,He J,et al.Deguelin,a selective silencer of the NPM1mutant,potentiates apoptosis and induces differentiation in AMLcells carrying the NPM1 mutation[J].Ann Hematol,2015,94(2):201-210.
[8]Stuwe T,Bley CJ,Thierbach K,et al.Architecture of the fungal nuclear pore inner ring complex[J].Science,2015,350(6256):56-64.
[9]Tung N,Battelli C,Allen B,et al.Frequency of mutations in individuals with breast cancer referred for BRCA1 and BRCA2 testing using next-generation sequencing with a 25-gene panel[J].Cancer,2015,121(1):25-33.
[10]Easton DF,Pharoah PD,Antoniou AC,et al.Gene-panel sequencing and the prediction of breast-cancer risk[J].New Engl J Med,2015,372(23):2243-2257.
[11]FACS JELM,Wecsler JS,Ph D MFPM,et al.Molecular markers for breast cancer diagnosis,prognosis and targeted therapy[J].J Surg Oncol,2015,111(1):81-90.
[12]Govindan R.Overcoming resistance to targeted therapy for lung cancer[J].New Engl J Med,2015,372(18):1760-1761.
[13]Mukherjee N,Schwan JV,Fujita M,et al.Alternative treatments for melanoma:targeting BCL-2 family members to de-bulk and kill cancer stem cells[J].J Invest Dermatol,2015,135(9):2155-2161.
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[15]Lecuit T,Yap AS.E-cadherin junctions as active mechanical integrators in tissue dynamics[J].Nat Cell Biol,2015,17(5):533-539.
[16]Kang DW,Yeol CC,Yong-Hee C,et al.Targeting phospholipase D1 attenuates intestinal tumorigenesis by controllingβ-catenin signaling in cancer-initiating cells[J].J Exp Med,2015,212(8):1219-1237.