Inhibition Of Hela Cell Growth And The Expression Of HIF-1α And VEGF By Antisense Oligodexynucleotide Targeting Hypoxia Inducible Factor 1α Gene

Background: Recently, antisense technology was generally applied to the research of tumour gene theraphy. With the development of genomics and proteomics . Some of target gene was screened out in the course of transformation and development of tumour. some author discover that HIF-1αmay be the key translator-factor to adapt the hypoxia condition for most cells. HIF-1αwas overexpression in multiple malignant neoplasm. HIF-1αcan modulate expression of multiple target gene like vascular endothelial growth factor(VEGF),erythropoietin(EPO) and sustain energy metabolism of neoplasm, contribute angiogenesis, promote proliferation and metastasis of neoplasm. VEGF was important in endothelial growth and tumorigenesis. It was correlation between angiogenesis and growth and metastasis of tumour. So HIF-1αmay be potential target for tumour gene therapy.Objective: To study inhibitive effect of HIF-1αfully phosporothioated antisense oligodeoxynucleotide (HIF-1αASODN) on the expression of HIF-1αand VEGF and proliferation of cervical cancer Hela cell line in vitro. This study will help to identify HIF-1αas a new target for gynecologic tumor gene therapy.Methods:1.Hela cell line were were transfected by ASODN and SODN with lipofectamineTM 2000 reagent. There were four groups in this experiment which was ASODN, SODN, lipsomal and control group.2.The proliferation activity of cervical cancer Hela cells was determined using methyl thiazolyl tetrazolium(MTT).3. The expression of HIF-1αand VEGF was detected by immunofluorescence, RT-PCR and western blot. HIF-1αgene silencing was investigated.4.Using SPSS11.5 software to assay the data.Results:1.The inhibitory rates on proliferation of Hela cells transfected by ASODN were significantly higher compared with those of control, SODN, lipofectamine TM2000 and blank group(all P < 0. 05) with dose-dependent .2.The mRNA and protein expression of HIF-1αand VEGF in cervical cancer Hela cells was significantly downregulated by HIF-1αASODN (all P < 0. 05) .The SODN and lipofectamine TM2000 showed no such effect( P>0.05). The results indicated ASODN inhibited expression of HIF-1αwith specificity.3.Indirect immunofluorescene analysis of the expression of HIF-1αand VEGF. HIF-1αwas staining in the nuclear and cytoplasm and VEGF was staining only in the cytoplasm.The fluorescence intensity was significantly subdued by HIF-1αASODN than other control groups. The result was detected that conformed to previous measure.Conclusion:1.HIF-1αwas staining in the nuclear and cytoplasm and VEGF was staining only in the cytoplasm.2.HIF-1αantisense oligodexynucleotide can obviously inhibit the proliferation and expression of HIF-1αand VEGF in Hela cells.3.HIF-1αASODN transfection might be an important new treatment for uterine cervix carcinoma.