[Influence of hypoxia inducible factor-1alpha on cervical cancer cell line HeLa in vitro].
OBJECTIVE
To explore the direct influence of hypoxia inducible factor-1alpha (HIF-1alpha) on the development of invasive cervical cancer and the possible molecular mechanism.


METHODS
Recombinant antisense targeting HIF-1alpha eukaryotic expression vector was constructed and transfected into cultured human cervical cancer cell line HeLa to reduce the expression of HIF-1alpha and its effect on cell proliferation, apoptosis, invasion and the cascade downstream gene expression of HIF-1alpha, including vascular endothelial growth factor (VEGF), glucose transport 1 (GLUT1) and multidrug resistance 1 (MDR1) genes was observed. The chemical method using CoCl(2) to induce hypoxia environment of growing cell was performed. Cells were divided into six groups, NN (normal non-transfected), NI (normal invalid transfected), NT (normal transfected), HN (hypoxia non-transfected), HI (hypoxia invalid transfected), and HT (hypoxia transfected). Methyl thiazolyl tetrazolium (MTT), flow cytometry, and Transwell methods were performed to evaluate the proliferation, invasion and apoptosis, and RT-PCR method was used to detect the gene expression of HIF-1alpha, VEGF, GLUT1 and MDR1.


RESULTS
After induction of hypoxia by CoCl(2), the change of gene expression of HIF-1alpha in HN (or HI) group compared to that in NN (or NI) group was not obvious (P > 0.05), but expression of VEGF, GLUT1 and MDR1 were all enhanced and overall proliferation was promoted, apoptosis inhibited [(11.46 +/- 0.28)% vs (29.27 +/- 0.18)%, (15.77 +/- 0.49)% vs (31.13 +/- 0.08)%], and transmembrane behavior enhanced [(37 +/- 12)% vs (26 +/- 7)%, (40 +/- 9)% vs (28 +/- 5)%], and the variations were significant (P < 0.05). On the contrary, transfection with pcDNA3.0/HIF-1alpha was companied by declined gene expression of HIF-1alpha (NT: 0.05 +/- 0.12, HT: 0.04 +/- 0.16), and all the variations were significant (P < 0.05).


CONCLUSIONS
HIF-1alpha may participate in malignant biological behaviors of cervical cancer such as anti-apoptosis, accelerating proliferation, increasing supply of blood and energy, increased resistance to chemotherapy through upregulation of its downstream genes. Suppression of HIF-1alpha expression in vitro can inhibit cervical cancer cell line.
Cheng
Yan-xiang
YX
Department of Obstetrics and Gynecology, Tongji Hospital of Tongji Medical College, Huazhong Science and Technology University, Wuhan 430032, China.
Pu
De-min
DM
Liu
Rong
R
Li
Tian
T
Yin
Ling
L
Ma
Ding
D
D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1 / Q000235:genetics; D017209:Apoptosis / Q000235:genetics / Q000502:physiology*; D015687:Cell Hypoxia; D049109:Cell Proliferation; D005260:Female; D005434:Flow Cytometry; D015972:Gene Expression Regulation, Neoplastic; D051272:Glucose Transporter Type 1 / Q000235:genetics; D006367:HeLa Cells; D006801:Humans; D051795:Hypoxia-Inducible Factor 1, alpha Subunit / Q000235:genetics / Q000502:physiology*; D016376:Oligonucleotides, Antisense / Q000235:genetics; D010957:Plasmids; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014162:Transfection; D002583:Uterine Cervical Neoplasms / Q000235:genetics / Q000473:pathology; D042461:Vascular Endothelial Growth Factor A / Q000235:genetics
D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1; D051272:Glucose Transporter Type 1; D051795:Hypoxia-Inducible Factor 1, alpha Subunit; D016376:Oligonucleotides, Antisense; D012333:RNA, Messenger; D042461:Vascular Endothelial Growth Factor A