KIF3B基因对三阴性乳癌细胞生物学特性和阿霉素化疗敏感性的影响

EFFECT OF THE KIF3B GENE ON THE BIOLOGICAL CHARACTERISTICS AND DOXORUBICIN CHEMOSENSITIVITY OF TRIPLE-NEGATIVE BREAST CANCER CELLS

目的研究KIF3B基因对三阴性乳癌MDA-MB-231细胞生物学特性和阿霉素化疗敏感性的影响。方法用Western blot技术检测KIF3B在人三阴性乳癌细胞系MDA-MB-231和MDA-MB-468中的表达,取KIF3B高表达细胞MDA-MB-231用于后续实验。用sh-NC质粒转染MDA-MB-231细胞作为对照组,用sh-KIF3B沉默质粒转染MDA-MB-231细胞作为实验组。应用Western blot方法检测基因沉默效率和基因沉默后上皮-间质转化(EMT)相关蛋白E-cadherin、基质金属蛋白酶2(MMP-2)和MMP-9表达变化,Transwell实验检测细胞迁移、侵袭能力变化,流式细胞术检测细胞周期变化,MTT实验检测细胞增殖能力和对阿霉素化疗敏感性的变化。结果KIF3B在三阴性乳癌细胞MDA-MB-231中高表达,而在MDA-MB-468中低表达(t=19.92,P<0.01)。沉默MDA-MB-231细胞KIF3B基因后,细胞迁移、侵袭数目明显减少(t=29.54、18.32,P<0.01),G_(0)/G_(1)期细胞比例降低而G_2/M期细胞比例增加(t=4.82、19.05,P<0.01),同时细胞增殖能力被显著抑制(F=7.56~270.09,P<0.01),对阿霉素化疗敏感性明显增加(F=26.37~167.11,P<0.01),E-cadherin表达升高(t=19.71,P<0.01),而MMP-2和MMP-9表达降低(t=26.57、16.11,P均<0.01)。结论沉默三阴性乳癌MDA-MB-231细胞KIF3B基因表达可以抑制细胞增殖、阻滞细胞周期并增加细胞对阿霉素化疗敏感性,并可以通过EMT抑制细胞迁移和侵袭。

Objective To investigate the effect of the KIF3B gene on the biological characteristics and doxorubicin chemosensitivity of triple-negative breast cancer MDA-MB-231 cells.Methods Western blot was used to measure the expression of KIF3B in human triple-negative breast cancer cell lines MDA-MB-231 and MDA-MB-468,and the MDA-MB-231 cells with high expression of KIF3B were selected for subsequent experiments.MDA-MB-231 cells transfected with sh-NC plasmid were established as control group,and those transfected with sh- KIF3B silencing plasmid were established as experimental group.Western blot was used to measure silencing efficiency and changes in the expression of the epithelial-mesenchymal transition(EMT)-related proteins E-cadherin,matrix metallopeptidase-2(MMP-2),and matrix metallopeptidase-9(MMP-9);Transwell assay was used to measure the changes in cell migration and invasion abilities;flow cytometry was used to observe the change in cell cycle;MTT assay was used to measure the changes in proliferative capacity and doxorubicin chemosensitivity.Results KIF3B showed high expression in triple-negative breast cancer MDA-MB-231 cells and low expression in MDA-MB-468 cells(t=19.92,P<0.01).After the KIF3B gene was silenced in MDA-MB-231 cells,there were significant reductions in the number of migrating and invading cells(t=29.54,18.32;P<0.01),a significant reduction in the proportion of cells in G_(0)/G_(1) phase,and a significant increase in the proportion of cells in G 2/M phase(t=4.82,19.05;P<0.01).Meanwhile,cell proliferation was significantly inhibited(F=7.56-270.09,P<0.01),and doxorubicin sensitivity was significantly increased(F=26.37-167.11,P<0.01).There were an increase in the expression of E-cadherin(t=19.71,P<0.01)and reductions in the expression of MMP-2 and MMP-9(t=26.57,16.11;P<0.01).Conclusion Silencing of the KIF3B gene in triple-negative breast cancer MDA-MB-231 cells can inhibit cell proliferation,cause cell cycle arrest,and increase the doxorubicin chemosensitivity of cells,and it can also inhibit cell migration and invasion through EMT.