Effects of KIF3A gene silencing and overexpression on proliferation, migration and invasion of human triple negative breast cancer cells

Wang Weilin; Wango Haib; Zhang Runze; Wang Xiao; Wang Jin; Lin Mei; Wang Chengqin

doi:10.3760/cma.j.issn.1001-9030.2019.12.014

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• 实验研究 •

KIF3A基因沉默和过表达对人三阴性乳腺癌细胞增殖、迁移及侵袭能力的影响

中华实验外科杂志, 2019,36(12) : 2173-2175. DOI: 10.3760/cma.j.issn.1001-9030.2019.12.014

摘要

目的

观察KIF3A基因沉默和过表达对人三阴性乳腺癌(TNBC)细胞MDA-MB-231增殖、迁移及侵袭能力的影响。

方法

通过慢病毒干扰技术对高表达KIF3A的人TNBC细胞株MDA-MB-231进行KIF3A基因沉默，通过脂质体转染法使KIF3A低表达的人TNBC细胞株MDA-MB-468 KIF3A基因过表达。应用蛋白质印迹法(Western blot)检测KIF3A基因沉默和过表达效率。应用平板克隆实验、Transwell迁移及侵袭实验检测KIF3A基因沉默和过表达对细胞增殖、迁移及侵袭能力的影响，应用SPSS 23.0统计软件分析。

结果

Western blot检测显示KIF3A沉默组(KIF3A-shRNA组)细胞中KIF3A的蛋白水平明显低于阴性对照组(Scr-shRNA组)，KIF3A过表达组(KIF3A-pEX组)细胞中KIF3A的蛋白水平明显高于阴性对照组(Vector组)；平板克隆实验结果显示KIF3A-shRNA组细胞克隆形成数目明显低于Scr-shRNA组[174.80±46.26比293.20±20.93，P<0.01]；KIF3A-pEX组细胞克隆形成数目明显高于Vector组[292.00±75.59比151.40±68.58，P<0.05]；Transwell迁移实验显示KIF3A-shRNA组穿膜细胞数/低倍视野明显低于Scr-shRNA组[(47.60±5.77)个比(161.40±20.16)个，P<0.01]；KIF3A-pEX组穿膜细胞数/低倍视野明显高于Vector组[(262.00±23.35)个比(155.00±29.15)个，P<0.01]；Transwell侵袭实验显示KIF3A-shRNA组穿膜细胞数/低倍视野明显低于Scr-shRNA组[(161.40±16.16)个比(281.00±19.77)个，P<0.01]；KIF3A-pEX组穿膜细胞数/低倍视野明显高于Vector组[(214.00±34.54)个比(125.40±19.94)个，P<0.01]。

结论

KIF3A基因沉默显著抑制TNBC细胞的增殖、迁移和侵袭能力，而KIF3A基因过表达可显著促进TNBC细胞的增殖、迁移和侵袭能力。

引用本文: 王伟淋, 王海波, 张润泽, 等. KIF3A基因沉默和过表达对人三阴性乳腺癌细胞增殖、迁移及侵袭能力的影响 [J] . 中华实验外科杂志,2019,36 (12): 2173-2175. DOI: 10.3760/cma.j.issn.1001-9030.2019.12.014

参考文献导出: Endnote NoteExpress RefWorks NoteFirst 医学文献王

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乳腺癌是女性最常见的恶性肿瘤和癌症死亡原因之一^[1]。三阴性乳腺癌(TNBC)是乳腺癌一个亚型，即雌激素受体(ER)、孕激素受体(PR)和人类表皮生长因子受体2(Her-2)均为阴性。且具有恶性程度高、侵袭性强、治疗棘手等特点^[2]。

贡献者信息

王伟淋

青岛大学基础医学院　266021

王海波

青岛大学附属医院乳腺外科　266003

张润泽

青岛大学基础医学院　266021

王笑

青岛大学基础医学院　266021

王瑾

青岛大学基础医学院　266021

林梅

青岛大学基础医学院　266021

王成勤

青岛大学基础医学院　266021

通信作者

王成勤

青岛大学基础医学院　266021

Email：wcq7618@126.com

关键词

KIF3A; 慢病毒干扰载体; 三阴性乳腺癌; 增殖; 迁移; 侵袭;

基金项目

国家自然科学基金（81672606）

利益冲突

利益冲突　所有作者均声明不存在利益冲突

历史

出版日期：2019-12-08

收稿日期：2019-07-11

Experimental Study

Effects of KIF3A gene silencing and overexpression on proliferation, migration and invasion of human triple negative breast cancer cells

Wang Weilin, Wango Haib, Zhang Runze, Wang Xiao, Wang Jin, Lin Mei, Wang Chengqin

Published 2019-12-08

Cite as Chin J Exp Surg, 2019,36(12): 2173-2175. DOI: 10.3760/cma.j.issn.1001-9030.2019.12.014

Abstract

Objective

To study the effects of KIF3A silencing and overexpression on tumor proliferation, migration and invasion in human triple negative breast cancer (TNBC) cell lines.

Methods

With lentivirus-mediated interference technology made KIF3A gene silence in TNBC MDA-MB-231 cells, KIF3A gene was transfected into TNBC MDA-MB-468 cells bv liposome method. Subsequently, Western blotting analysis was performed to detect protein expression of KIF3A in these cells. Colony-formation assay, transwell migration and invasion assay were applied to estimate proliferation, migration and invasion ability of cells.

Results

The expression of KIF3A protein in the KIF3A-shRNA group cells was significantly silenced compared with those in the Scr-shRNA group cells. The expression of KIF3A in the KIF3A-pEX group cells was increased compared with those in the Vector group cells. The colony numbers in the KIF3A-shRNA group were significantly lower than those in Scr-shRNA group (174.8±46.26 vs. 293.2±20.93, P<0.01). The colony numbers in the KIF3A-pEX group were significantly higher than those in Vector group cells (292.00±75.59 vs. 151.40±68.58, P<0.05). The result of the transwell migrated assay showed that the numbers of migrated cells in the KIF3A-shRNA group were significantly lower than those in Scr-shRNA group (47.60±5.77 vs. 161.40±20.16, P<0.01) and the numbers of migrated cells in the KIF3A-pEX group were significantly higher than those in Vector group (262.00±23.35 vs. 155.00±29.15, P<0.01). It was confirmed via the transwell invasion assay results that the cell’s invasive ability of KIF3A-shRNA group was progressively suppressed as compared with the Scr-shRNA group (161.40±16.16 vs. 281.00±19.77, P<0.01) and the cell’s invasive ability of KIF3A-pEX group was progressively promoted as compared with the Vector group (214.00±34.54 vs. 125.40±19.94, P<0.01).

Conclusion

Down-regulation of KIF3A in TNBC cell can inhibit the cell proliferation, migration and invasion. Overexpression of KIF3A in TNBC cell can promote the cell proliferation, migration and invasion. KIF3A might hold potential as a therapeutic drug target for human TNBC.

Key words:

KIF3A; Short hairpin RNA; Triple negative breast cancer; Proliferation; Migration; Invasion

Contributor Information

Wang Weilin