关键词: 宫颈癌, RNA干扰, 低氧诱导因子1α, 定植, 侵袭

Abstract: Objective To construct 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 designed based on HIF-1αgene sequence and was cloned into pSilencer2.1-U6-neo vector. The resultant plasmid was transfected into HeLa cells with Lipofectamine 2000. The cells were incubated under hypoxic condition. The HIF-1αprotein and mRNA were detected by Western blot and real time 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-1αin HeLa cells decreased, and the number of colony formation in soft agar and cells penetrating matrigel also decreased under hypoxic condition. Conclusions The shRNA expressing plasmid targeting HIF-1αcould suppress the location and metastasis of cervical carcinoma cells under hypoxic condition.

Key words: Cervical carcinoma, RNA interference, Hypoxia-inducible factor 1α, Location, Metastasis