Effects of hypoxia inducible factor-1alpha siRNA
on the invasion of human Hela cells and expression of related proteins

doi:10.1007/s11684-009-0060-3

Front. Med.

2009, Vol. 3

Issue (3) : 303-308 https://doi.org/10.1007/s11684-009-0060-3

Research articles

Effects of hypoxia inducible factor-1alpha siRNA on the invasion of human Hela cells and expression of related proteins

Bin YANG MS ¹, Xianglin YUAN ², Yanmei ZOU ², Qingsong XI ², Guoxian LONG ², Qiang FU ², Guangyuan HU MM ²,

1.Center of Oncology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, China；First Ward of Internal Medicine, Tumor Hospital of Hubei Province, Wuhan 430067, China; 2.Center of Oncology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, China;

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Abstract The effects of hypoxia on the invasion and the related protein expression of Hela cells and the role of hypoxia inducible factor-1α (HIF-1α) were investigated. The Hela cells were divided into three groups, namely, H₀ (non-transfected Hela cells), H₁ (pGenesil-1 empty plasmid-transfected Hela cells), and H₂ (HIF-1α-shRNA plasmid-transfected Hela cells), and were cultured under hypoxia (1% O₂) and normoxia for 48h. The expression of HIF-1α, E-cadherin, β-catenin, and actin was detected using Western blot. The scratch test and the invasion assay were applied to examine the invasion in each group. The expression of HIF-1α, E-cadherin, and β-catenin in tumor grafts was assayed immunohistochemically. Western blot results revealed that the bands of HIF-1α, E-cadherin, β-catenin, and actin proteins were detected in the H₀ and H₁ groups under hypoxia for 48h. The expression of E-cadherin, β-catenin, and actin was detected in the H₂ group under hypoxia for 48h, and normoxia. In the H₀, H₁, and H₂ groups under normoxia, and the H₂ group under hypoxia for 48h, no expression of HIF-1α was detectable. The scratch test showed that the invasive ability in the H₂ group was significantly alleviated. Immunohistochemically, it was found that there was a significant difference in the expression of HIF-1α, E-cadherin, and β-catenin between the H₁ and H₂ groups (P<0.05), but the difference was not significant between the H₀ and H₁ groups. It was concluded that the effects of hypoxia on the invasion of human cervical cancer Hela cells and the expression of related proteins (E-cadherin, β-catenin, and actin) depend on HIF-1α.

Keywords hypoxia actin;β-catenin E-cadherin invasion

Issue Date: 05 September 2009

Cite this article:

Bin YANG MS,Yanmei ZOU,Xianglin YUAN, et al. Effects of hypoxia inducible factor-1alpha siRNA on the invasion of human Hela cells and expression of related proteins[J]. Front. Med., 2009, 3(3): 303-308.

URL:

https://academic.hep.com.cn/fmd/EN/10.1007/s11684-009-0060-3
https://academic.hep.com.cn/fmd/EN/Y2009/V3/I3/303

	Fidler I J. The pathogenesis of cancer metastasis: the seed and oil hypothesisrevisited. Nat Rev Cancer, 2003, 3(6): 453―458 doi: 10.1038/nrc1098
	Rivard N. Phosphatidylinositol3-kinase: a key regulator in adherent junction formation and function. Front Biosci, 2009, 14: 510―522 doi: 10.2741/3259
	Sonq Y, Wang W, Qu X, Sun S. Effects ofhypoxia inducible factor-1alpha (HIF-1alpha) on the growth & adhesionin tongue squamous cell carcinoma cells. Indian J Med Res. 2009, 129(2): 154―163
	Song I S, Wang A G, Yoon S Y, Kim J M, Kim J H, Lee D S, Kim N S. Regulation of glucose metabolism-relatedgenes and VEGF by HIF-1alpha and HIF-1beta, but not HIF-2alpha, ingastric cancer. Exp Mol Med, 2009, 41(1): 51―58 doi: 10.3858/emm.2009.41.1.007
	Yuan X L, Yu S L, Zhuang L, Xie T, Yang B, Dong L L, Xiong H. Interference of HIF-1αby RNA to enhance radio sensitivity of HeLa cell tumor transplantedin BALB/c mice. Zhongguo Aizheng Zazhi, 2007, 17(4): 303―306 (in Chinese)
	Sambrook J, Russell D W. The Condensed Protocols fromMolecular Cloning: a Laboratory Manual (in Chinese, trans. Huang Peitang). 3rd ed. Beijing: Science Press, 2002
	Escuin D, Simons J W, Giannakakou P. Exploitation of the HIF axis for Cancer Therapy. Cancer Biol Ther, 2004, 3(7): 608―611
	Fidler I J, Kim S J, Langley R R. The role of the organ microenvironment in the biologyand therapy of cancer metastasis. J CellBiochem, 2007, 101(4): 927―936 doi: 10.1002/jcb.21148
	Luo Y, He D L, Ning L. Hypoxia-inducible factor-1alpha induces the epithelialmesenchymal transition of human prostatecancer cells. Chin Med, 2006, 119(9): 713―718
	Ma J, Zhang L, Ru G Q, Zhao Z S, Xu W J. Upregulation of hypoxia inducible factor1alpha mRNA is associated with elevated vascular endothelial growthfactor expression and excessive angiogenesis and predicts a poor prognosisin gastric carcinoma. World J Gastroenterol, 2007, 13(11): 1680―1686
	Cardone R A, Bellizzi A, Busco G, Weinman E J, Dell'Aquila M E, Casavola V, Azzariti A, Mangia A, Paradiso A, Reshkin S J. The NHERF1 PDZ2 domain regulates PKA-RhoA-p38-mediated NHE1 activationand invasion in breast tumor cells. MolBiol Cell, 2007, 18(5): 1768―1780 doi: 10.1091/mbc.E06-07-0617
	Wojciak-Stothard B, Tsang L Y, Paleolog E. Hall S M, Harworth S G. Rac1 and RhoA as regulatorsof endothelial phenotype and barrier function in hypoxia-induced neonatalpulmonary hypertension. Am J Physiol LungCell Mol Physiol, 2006, 290(6): L1173―1182 doi: 10.1152/ajplung.00309.2005
	Aono Y, Ariyoshi H, Tsuji Y, Ueda A, Tokunaga M, Sakon M, Monden M. Localized activation of m-calpainin human umbilical vein endothelial cells upon hypoxia. Thtomb Res, 2001, 102(4): 353―361 doi: 10.1016/S0049-3848(01)00238-9
	Nelson W J, Nusse R. Convergence of Wnt, beta-catenin,and cadherin pathways. Science, 2004, 303(5663): 1483―1487 doi: 10.1126/science.1094291
	Menez J, Le Maux Chansac B, Dorothée G, Vergnon I, Jalil A, Carlier M F, Chouaib S, Mami-Chouaib F. Oncogene, 2004, 23 (15): 2630―2639 doi: 10.1038/sj.onc.1207347
	Calvisi D F, Ladu S, Conner E A, Factor V M, Thorgeirsson S S. Disregulation of E-cadherinin transgenic mouse models of liver cancer. Lab Invest, 2004, 84(9): 1137―1147 doi: 10.1038/labinvest.3700147
	Sun S, Ning X, Zhang Y, Lu Y, Nie Y, Han S, Liu L, Du R, Xia L, He L, Fan D. Hypoxia-induciblefactor-1alpha induces Twist expression in tubular epithelial cellssubjected to hypoxia, leading to epithelial-to-mesenchymal transition. Kidney Int, 2009, 75(12): 1278―1287 doi: 10.1038/ki.2009.62
	Yan W, Fu Y, Tian D, Liao J, Liu M, Wang B, Xia L, Zhu Q, Luo M. PI3 kinase/Aktsignaling mediates epithelial-mesenchymal transition in hypoxic hepatocellularcarcinoma cells. Biochem Biophys Res Commun, 2009, 2009, 382(3): 631―636
	Hrroshi I, Hideyuki S, Masatoshi H. Expression of hypoxia inducible factor-1α predictsmetastasis-free survical after radiation therapy alone in stage IIIBcervical squamous cell carcinoma. Int JRadiat Oncol Biol Phys, 2004, 50(4): 865―872
	Ishikawa H, Sakurai H, Hasegawa M, Mitsuhashi N, Takahashi M, Masuda N, Nakajima M, Kitamoto Y, Saitoh J, Nakano T. Expressionof Hypoxic-inducible factor-l alpha predicts metastasis-free survivalafter radiation therapy alone in stage ШB cervical squamous cellcarcinoma. Internat J Radia Oncol Phys, 2004, 60(2): 513―521 doi: 10.1016/j.ijrobp.2004.03.025
	Bachtiary B, Schindl M, Potter R, Dreier B, Knocke T H, Hainfellner J A, Horvat R, Birner P. Overexpression of hypoxiainducible factor-1alpha indicates diminished response to radiotherapyand unfavorable prognosis in patients receiving radiotherapy for cervicalcancer. Clin Cancer Res, 2003, 9(6): 2234―2240

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