Xi bao yu fen zi mian yi xue za zhi = Chinese journal of cellular and molecular immunology · 2016
[Overexpression of IL-8 promotes migration of BT549 breast cancer cells].
Аннотация на английскомСтатьяПоддержка исследования: вне правительства СШАНазвание и аннотация: Перевод в очереди
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Deng Fang, Wang Jing, Fan Mengtian, Guo Yangliu, Li Ya, Shi Qiong
Аннотация
OBJECTIVE: To construct a recombinant adenovirus vector containing IL-8 gene and observe its effect on the proliferation, cell cycle and migration of BT549 breast cancer cells.
METHODS: IL-8 gene was amplified by PCR using the cDNA from 143B bone sarcoma cells and inserted into shuttle plasmid pAdTrack-TO4. The recombinant shuttle plasmid pAdTrack-TO4-IL-8 was digested by PmeI and then transformed to AdEasier competent cells. The obtained recombinant adenovirus plasmid pAdIL-8 was digested by PacI, and then transfected to HEK293 cells for package and amplification by Lipofectamine(TM) 2000. The titer was tested by dilution assay. The expression of IL-8 mRNA and protein in BT549 cells was detected by reverse transcription PCR and ELISA, respectively. Effect of IL-8 overexpression on proliferation, cell cycle and migration in BT549 cells was respectively investigated by MTT assay, flow cytometry and wound-healing test.
RESULTS: PCR and DNA sequence analysis verified the recombinant shuttle plasmid pAdTrack-TO4-IL-8. Restriction enzymes PacI confirmed the recombinant adenovirus plasmid pAdIL-8. IL-8 was overexpressed in BT549 cells after AdIL-8 infection. Overexpression of IL-8 promoted BT549 cell migration and arrested the cell cycle in the S phase, but it made no significant difference in the proliferation of BT549 cells.
CONCLUSION: IL-8 overexpression can promote migration of BT549 breast cancer cells.
METHODS: IL-8 gene was amplified by PCR using the cDNA from 143B bone sarcoma cells and inserted into shuttle plasmid pAdTrack-TO4. The recombinant shuttle plasmid pAdTrack-TO4-IL-8 was digested by PmeI and then transformed to AdEasier competent cells. The obtained recombinant adenovirus plasmid pAdIL-8 was digested by PacI, and then transfected to HEK293 cells for package and amplification by Lipofectamine(TM) 2000. The titer was tested by dilution assay. The expression of IL-8 mRNA and protein in BT549 cells was detected by reverse transcription PCR and ELISA, respectively. Effect of IL-8 overexpression on proliferation, cell cycle and migration in BT549 cells was respectively investigated by MTT assay, flow cytometry and wound-healing test.
RESULTS: PCR and DNA sequence analysis verified the recombinant shuttle plasmid pAdTrack-TO4-IL-8. Restriction enzymes PacI confirmed the recombinant adenovirus plasmid pAdIL-8. IL-8 was overexpressed in BT549 cells after AdIL-8 infection. Overexpression of IL-8 promoted BT549 cell migration and arrested the cell cycle in the S phase, but it made no significant difference in the proliferation of BT549 cells.
CONCLUSION: IL-8 overexpression can promote migration of BT549 breast cancer cells.