Weilin Wang, Haib Wango, Runze Zhang, Xiao Wang, Jin Wang, Meisui Lin, Chengqin Wang
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Colony-formation assay, transwell migration and invasion assay were applied to estimate proliferation, migration and invasion ability of cells. \n \n \nResults \nThe 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). \n \n \nConclusion \nDown-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. \n \n \nKey words: \nKIF3A; Short hairpin RNA; Triple negative breast cancer; Proliferation; Migration; Invasion","PeriodicalId":10065,"journal":{"name":"中华实验外科杂志","volume":"36 1","pages":"2173-2175"},"PeriodicalIF":0.0000,"publicationDate":"2019-12-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"Effects of KIF3A gene silencing and overexpression on proliferation, migration and invasion of human triple negative breast cancer cells\",\"authors\":\"Weilin Wang, Haib Wango, Runze Zhang, Xiao Wang, Jin Wang, Meisui Lin, Chengqin Wang\",\"doi\":\"10.3760/CMA.J.ISSN.1001-9030.2019.12.014\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"Objective \\nTo study the effects of KIF3A silencing and overexpression on tumor proliferation, migration and invasion in human triple negative breast cancer (TNBC) cell lines. \\n \\n \\nMethods \\nWith 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. \\n \\n \\nResults \\nThe 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). \\n \\n \\nConclusion \\nDown-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. 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引用次数: 0
摘要
目的研究KIF3A沉默和过表达对人三阴性乳腺癌(TNBC)细胞系肿瘤增殖、迁移和侵袭的影响。方法采用慢病毒介导干扰技术使KIF3A基因沉默TNBC MDA-MB-231细胞，脂质体法将KIF3A基因转染TNBC MDA-MB-468细胞。随后，采用Western blotting分析检测KIF3A蛋白在这些细胞中的表达。采用菌落形成试验、跨井迁移和侵袭试验评估细胞的增殖、迁移和侵袭能力。结果与Scr-shRNA组细胞相比，KIF3A蛋白在KIF3A- shrna组细胞中表达明显沉默。与Vector组细胞相比，KIF3A- pex组细胞中KIF3A的表达增加。KIF3A-shRNA组的菌落数显著低于Scr-shRNA组(174.8±46.26 vs 293.2±20.93,P<0.01)。kif3 - pex组细胞集落数显著高于Vector组(292.00±75.59 vs. 151.40±68.58,P<0.05)。transwell迁移实验结果显示，kif3 - shrna组的迁移细胞数显著低于Scr-shRNA组(47.60±5.77 vs. 161.40±20.16,P<0.01)， kif3 - pex组的迁移细胞数显著高于Vector组(262.00±23.35 vs. 155.00±29.15,P<0.01)。跨井侵袭实验结果证实，与Scr-shRNA组相比，kif3 - shrna组细胞的侵袭能力被逐渐抑制(161.40±16.16比281.00±19.77,P<0.01);与Vector组相比，kif3 - pex组细胞的侵袭能力被逐渐增强(214.00±34.54比125.40±19.94,P<0.01)。结论下调KIF3A可抑制TNBC细胞的增殖、迁移和侵袭。KIF3A在TNBC细胞中过表达可促进细胞增殖、迁移和侵袭。KIF3A可能具有作为人类TNBC治疗药物靶点的潜力。关键词:KIF3A;短发夹RNA;三阴性乳腺癌;扩散;迁移;入侵
Effects of KIF3A gene silencing and overexpression on proliferation, migration and invasion of human triple negative breast cancer cells
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
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