Cx43对乳腺癌细胞侵袭和迁移及细胞间隙通讯的影响

摘要

To explore the effects of Cx43 on the invasion,migration and gap junctional intercellular communication(GJIC)of breast cancer. Methods:The Cx43 plasmid,inhibitors and negative controls were transfected into human breast cancer cell line MDA - MB - 231 by liposome. The motility of cells was evaluated by adhesion and Transwell assay. MMP - 2,MMP - 9 and BRMS - 1 were detected by both qPCR and Western blot. A scrape - loading dye transfer assay was used to evaluate the function of GJIC. Results:Compared to the wild type,adhesion ratio of Cx43 siRNA group(0. 43 ± 0. 07)was significantly lower(t = 20. 801,P = 0. 002),and that of Cx43 plasmid group (1. 63 ± 0. 26)was higher(t = 5. 917,P = 0. 027)by adhesion assay. Transwell tests showed the migration and inva-sion of Cx43 plasmid group was significantly increased( invasion:1. 25 ± 0. 04,t = 16. 339,P = 0. 004;migration:1. 33 ± 0. 02,t = 22. 770,P = 0. 002),but that of Cx43 siRNA group was significantly reduced( migration:0. 84 ± 0. 04,t = 11. 139,P = 0. 008;invasion:0. 79 ± 0. 04,t = 5. 743,P = 0. 029). The expression of MMP - 2 and MMP - 9 were enhanced and the expression of BRMS - 1 was reduced in Cx43 plasmid group detected by both qPCR and west-ern blot and vice versa in Cx43 siRNA group. Compared to the wild type,GJIC was enhanced in Cx43 plasmid group and reduced in Cx43 siRNA group by a scrape - loading dye transfer assay. Conclusion:Cx43 might have a positive control on invasion and migration of breast cancer cell by GJIC.%目的：探讨 Cx43对乳腺癌侵袭、迁移以及细胞间隙通讯功能的影响。方法：用脂质体转染法将 Cx43质粒、Cx43 siRNA 及阴性对照转染入人乳腺癌细胞株 MDA - MB -231。细胞黏附实验、Transwell 试验检测细胞黏附、侵袭和迁移能力。分子生物学方法检测 MMP -2、MMP -9、BRMS -1的表达。荧光染料传输实验检测不同 Cx43表达的细胞间隙通讯的功能。结果：细胞黏附实验结果表明相较于野生型细胞，Cx43 siRNA 组黏附能力（0.43±0.07）降低（t =20.801，P =0.002），Cx43质粒组黏附能力（1.63±0.26）增高（t =5.917，P =0.027）。Transwell 实验表明相较野生组，Cx43质粒组侵出小室的细胞数增加（侵袭：1.25±0.04，t =16.339，P =0.004；迁移：1.33±0.02，t =22.770，P =0.002），而 Cx43 siRNA 组侵出小室的细胞数减少（迁移：0.84±0.04，t =11.139，P =0.008；侵袭：0.79±0.04，t =5.743，P =0.029）。分子生物学检测发现 Cx43质粒组 MMP -2、MMP -9表达增高，BRMS -1表达降低，而 Cx43 siRNA 组 MMP -2、MMP -9表达降低，BRMS -1表达增高。染料传输实验证实相较野生组传输能力，Cx43质粒组传输作用增强；Cx43 siRNA 组细胞间的传输作用减弱。结论：Cx43对乳腺癌细胞侵袭及迁移能力存在正调控。这一调控能力可能通过 GJIC 实现。