In vitro effects of hypoxia-inducible factor 1alpha on the biological characteristics of the SiHa uterine cervix cancer cell line.
BACKGROUND
Hypoxia-inducible factor 1alpha (HIF-1alpha) regulates the transcription of many genes involved in key aspects of cancer biology. The aim of our study was to explore the effects of HIF-1alpha on the biological characteristics of the uterine cervix cancer (UCC) cell line SiHa, such as proliferation, apoptosis, and migration under normoxia and hypoxia.


METHODS
Full-length HIF-1alpha (fL HIF-1alpha) and dominant-negative HIF-1alpha (dn HIF-1alpha) were transfected into UCC SiHa cells. The expression of HIF-1alpha and its targets such as vascular endothelial growth factor (VEGF), CXC chemokine receptor 4 (CXCR4), and human growth and transformation-dependent protein (HGTD-P) was detected by immunocytochemistry, Western blot, and semiquantitative reverse transcription-polymerase chain reaction. Cell proliferation, apoptosis, and migration were surveyed by methyl thiazolyl tetrazolium assay, terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick-end labeling staining, and scratching test.


RESULTS
The expression of HIF-1alpha increased in fL HIF-1alpha but not in dn HIF-1alpha SiHa cells. Consistently, the expression of HIF-1alpha target genes such as VEGF, CXCR4, and HGTD-P increased in fL HIF-1alpha-transfected SiHa cells but decreased in dn HIF-1alpha-transfected SiHa cells. The UCC cells transfected with fL HIF-1alpha had increased cellular proliferation and migration. However, the inhibition of HIF-1alpha through dn HIF-1alpha attenuated cell proliferation and migration under both normoxia and hypoxia.


CONCLUSIONS
Hypoxia-inducible factor 1alpha affects the proliferation, apoptosis, and migration of UCC SiHa cells in part by regulating the expression of its target genes such as VEGF, HGTD-P, and CXCR4. Targeting HIF-1alpha may be a promising strategy for molecular therapy for UCC.
Tang
Binzhi
B
Department of Pediatrics, West China Women and Children's Hospital, Sichuan University, Chengdu, China.
Qu
Yi
Y
Zhao
Fengyan
F
Mao
Meng
M
Tang
Jun
J
Li
Xihong
X
Ferriero
Donna
D
Mu
Dezhi
D
D016587:Antimutagenic Agents / Q000494:pharmacology; D017209:Apoptosis; D015153:Blotting, Western; D002448:Cell Adhesion; D002465:Cell Movement; D049109:Cell Proliferation; D003035:Cobalt / Q000494:pharmacology; D005260:Female; D005786:Gene Expression Regulation / Q000502:physiology; D006801:Humans; D051795:Hypoxia-Inducible Factor 1, alpha Subunit / Q000502:physiology*; D007150:Immunohistochemistry; D066298:In Vitro Techniques; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism; D024101:Mitochondrial Proteins / Q000235:genetics / Q000378:metabolism; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D019718:Receptors, CXCR4 / Q000235:genetics / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014162:Transfection; D014407:Tumor Cells, Cultured; D002583:Uterine Cervical Neoplasms / Q000378:metabolism* / Q000473:pathology*; D042461:Vascular Endothelial Growth Factor A / Q000235:genetics / Q000378:metabolism
D016587:Antimutagenic Agents; C527276:CXCR4 protein, human; C486647:FAM162A protein, human; C497442:HIF1A protein, human; D051795:Hypoxia-Inducible Factor 1, alpha Subunit; D008565:Membrane Proteins; D024101:Mitochondrial Proteins; D012333:RNA, Messenger; D019718:Receptors, CXCR4; C467484:VEGFA protein, human; D042461:Vascular Endothelial Growth Factor A; D003035:Cobalt; C018021:cobaltous chloride