KCNQ1OT1基因敲除联合鸦胆子素D对乳腺癌MDA-MB-231细胞增殖、迁移及侵袭的影响

Effects of KCNQ1OT1 Gene Knockout Combined with Bruceine D on Proliferation,Migration,and Invasion of Breast Cancer MDA-MB-231 Cells

目的探讨KCNQ1OT1基因敲除联合鸦胆子素D对乳腺癌MDA-MB-231细胞生物学行为的影响及其机制。方法CCK8、划痕和Transwell侵袭实验分别检测鸦胆子素D和siKCNQ1OT1对MDA-MB-231细胞活力、迁移和侵袭能力的影响;qRT-PCR检测鸦胆子素D、siKCNQ1OT1对MDA-MB-231细胞中KCNQ1OT1表达的影响;Western blot法检测EMT相关蛋白及CDC42、p-MKK7、MKK7蛋白的表达情况。结果鸦胆子素D和si KCNQ1OT1处理的MDA-MB-231细胞活力、迁移和侵袭能力受到显著抑制,且二者联用时抑制作用更强(均P<0.05);鸦胆子素D处理后KCNQ1OT1在MDA-MB-231细胞中的表达下调(均P<0.05);鸦胆子素D联合si KCNQ1OT1处理组MDAMB-231细胞中CDC42、p-MKK7、N-c adherin和Vimentin的表达显著下调,E-cadherin的表达上调(均P<0.05)。结论鸦胆子素D联合si KCNQ1OT1抑制人乳腺癌MDA-MB-231细胞增殖、迁移、侵袭和EMT,其分子机制可能与CDC42/MKK7信号通路的阻断有关。

Objective To explore the effect of KCNQ1OT1 gene knockout combined with bruceine D on the proliferation,migration,and invasion of breast cancer MDA-MB-231 cells.Methods Cell Counting Kit-8,wound healing,and Transwell invasion assay were used to detect the effects of bruceine D and siKCNQ1OT1 on the viability,migration,and invasion of MDA-MB-231 cells.Effect of bruceine D and siKCNQ1OT1 on the expression of KCNQ1OT1 in MDA-MB-231 cells was detected by qRT-PCR.Western blot was used to detect the effect of bruceine D and siKCNQ1OT1 on the expression of EMT-related proteins and CDC42,p-MKK7,MKK7 proteins in MDA-MB-231 cells.Results Bruceine D and siKCNQ1OT1 could significantly inhibit the viability,migration,and invasion of MDA-MB-231 cells,and the inhibitory effect was enhanced when they were combined(all P<0.05);bruceine D downregulated the expression of KCNQ1OT1 in MDA-MB-231 cells(all P<0.05);bruceine D combined with siKCNQ1OT1 significantly decreased CDC42,p-MKK7,N-cadherin,and Vimentin expression in MDA-MB-231 cells and increased the expression of E-cadherin(all P<0.05).Conclusion Bruceine D combined with siKCNQ1OT1 significantly inhibit the proliferation,migration,invasion,and EMT of human breast cancer MDA-MB-231 cells,and its molecular mechanism may be related to the blocking of CDC42/MKK7 signaling pathway.