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

龙凤; 赵玉; 黄勇; 刘晓燕; 周旋; 李雪; 叶海琳

doi:10.3971/j.issn.1000-8578.2023.23.0456

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

摘要

摘要:
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siKCNQ1OT1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siKCNQ1OT1处理组MDA-MB-231细胞中CDC42、p-MKK7、N-cadherin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siKCNQ1OT1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。

Abstract:
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.

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