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目的探讨缝隙接蛋白(Cx)在钙离子介导的乳腺癌细胞转移和侵袭中的作用。方法选用不影响细胞生长的缝隙连接蛋白阻断剂,辛醇100μmol/L处理高转移MDA-MB-231细胞和低转移MCF-7细胞。倒置相差显微镜下观察细胞形态,激光扫描共焦显微镜下观察Cx43的位置、微丝纤维的排列和细胞内钙离子浓度变化,划痕和侵袭实验观察细胞的转移情况。结果辛醇处理细胞后,细胞的生长状态由大面积片状生长转变为单个或少数几个团状的独立生长;Cx43蛋白的形成和表达位置虽无改变,但相邻细胞间微丝纤维排列的平行同向性显著性降低;MDA-MB-231细胞穿过基底膜成胶和Transwell小室基底面的细胞数显著低于对照组;此外,细胞内钙离子浓度强度显著性降低,钙离子螯合剂(EGTA)处理显著性加剧辛醇对细胞转移和侵袭能力的抑制。但是,上述现象在低转移MCF-7细胞中效果不明显。结论缝隙连接蛋白阻断剂在干扰乳腺癌细胞Cx功能活性,而不影响Cx形成的情况下,能够显著性抑制高转移乳腺癌的恶性进展和侵袭转移,此机制与细胞内钙离子浓度降低有关。
Objective To investigate the role of Cx in Ca 2+ -mediated metastasis and invasion of breast cancer cells. Methods Gap junctional protein blockers were used to inhibit cell growth. Octamyl alcohol (100 μmol / L) was used to treat high metastatic MDA-MB-231 cells and low metastatic MCF-7 cells. Cell morphology was observed under an inverted phase contrast microscope. The location of Cx43, the arrangement of fibrillar fibers and the intracellular calcium concentration were observed under laser scanning confocal microscope. Scratches and invasion experiments were performed to observe the cell migration. Results After the cells were treated with octanol, the growth of the cells changed from large-area sheet-like growth to single or small clusters. The formation and expression of Cx43 protein did not change, but the arrangement of microfilaments in adjacent cells In the same direction; the number of MDA-MB-231 cells passing through the basement membrane and transwell basal cell surface was significantly lower than that of the control group; in addition, the intensity of intracellular calcium concentration was significantly reduced, Mixture (EGTA) treatment significantly aggravated the inhibition of octanol on cell metastasis and invasiveness. However, the above phenomenon is not obvious in low-metastatic MCF-7 cells. Conclusions The gap junction protein antagonist can significantly inhibit the malignant progression and invasion and metastasis of highly metastatic breast cancer by interfering with the Cx functional activity of breast cancer cells without affecting the formation of Cx, which is related to the decrease of intracellular calcium concentration .