Establishment of MDA-MB-231 cell line stably expressing inducible sh RNA targeting PLK1 and effects of PLK1 knockdown on cell proliferation of MDA-MB-231 cells
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Establishment of MDA-MB-231 cell line stably expressing inducible sh RNA targeting PLK1 and effects of PLK1 knockdown on cell proliferation of MDA-MB-231 cells
Journal of Chongqing Medical UniversityVol. 42, Issue 11, Pages: 1400-1405(2017)
Xue Hui, Guo Xin, Li Na, et al. Establishment of MDA-MB-231 cell line stably expressing inducible sh RNA targeting PLK1 and effects of PLK1 knockdown on cell proliferation of MDA-MB-231 cells. [J]. Journal of Chongqing Medical University 42(11):1400-1405(2017)
DOI:
Xue Hui, Guo Xin, Li Na, et al. Establishment of MDA-MB-231 cell line stably expressing inducible sh RNA targeting PLK1 and effects of PLK1 knockdown on cell proliferation of MDA-MB-231 cells. [J]. Journal of Chongqing Medical University 42(11):1400-1405(2017) DOI: 10.13406/j.cnki.cyxb.001400.
Establishment of MDA-MB-231 cell line stably expressing inducible sh RNA targeting PLK1 and effects of PLK1 knockdown on cell proliferation of MDA-MB-231 cells
Objective:To establishMDA-MB-231 cell line stably expressing inducible small hairpin RNA(shRNA)targeting polo-like kinase 1(PLK1)
to explore the effects of PLK1 knockdown on cell proliferation and cell cycle in MDA-MB-231 cells as well as to explore its possible mechanism. Methods:Two oligonucleotides targeting PLK1 were synthesized and cloned into lentivirus expression plasmid p LVTHM according to si RNA principle(named PLK1 shRNA). MDA-MB-231 cells were infected at first with lentivirus prepared by p LV-t TR/KRAB-Red empty vector
and then infected with another lentivirus prepared by PLK1 shRNA recombinant plasmid.MDA-MB-231 cells infected with aforementioned lentivirus were treated with doxycycline(DOX)for 96 hours
the expression of PLK1 m RNA and protein was determined by real time PCR(q RT-PCR)and Western blot respectively. MTT method was used to determine cell proliferation and flow cytometry and PI staining were used to detect cell cycle. Results :The lentivirus expression plasmids containing PLK1 shRNA were constructed successfully
and MDA-MB-231 cells were infected with the lentivirus prepared by the aforementioned plasmids. MDA-MB-231 cell line expressing stably inducible shRNA targeting PLK1 was established successfully and PLK1 expression levels were decreased obviously at m RNA and protein levels after induction by DOX(P<0.01). PLK1 knockdown could obviously inhibit the proliferation of MDA-MB-231 cells and arrest the cell cycle in G2/M phase(P<0.05). Conclusions:PLK1 knockdown induced by DOX could inhibit MDA-MB-231 cell proliferation and arrest cell cycle in G2/M phase
its possible mechanism may involve in ERK1/2/Fra1/ZEB1 signal pathway.