Cx43对乳腺癌细胞侵袭和迁移及细胞间隙通讯的影响

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。

Effect of Cx43 on the invasion and migration and Gap jjnctional intercellular communica-tion in breast cancer cell

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.