Effects of RNA interference targeting HIF-1α on location and metastasis in HeLa cells / 基础医学与临床

ABSTRACT

Objective To construct an eukaryotic vector expressing short hairpin RNA(shRNA) of HIF-1α,and to observe its effects on location and metastasis of HeLa cells under hypoxic condition.Methods shRNA templates was developed based on HIF-1α gene sequence and then cloned into pSilencer2.1-U6-neo vector.The resultant plasmid was transfected into HeLa cells with Lipofectamine 2000.The cells were incubated in hypoxic condition.The HIF-1α protein and mRNA were detected by Western blot and RT-PCR.The colony formation assay and Transwell cabin assay were performed to measure the colony formation and metastasis.Results The plasmid pSilencer2.1-U6-neo-HIF-1α was successfully constructed and transfected into HeLa cells.The expression of HIF-1α in HeLa cells decreased,and the number of colony formation in soft agar and cells penetrating matrigel also decreased under hypoxic condition.Conclusion The shRNA expressing plasmid targeting at HIF-1α may suppress the location and metastasis of cervical carcinoma cells under hypoxic condition.