| Original Article |
|
|
|
|
| Antagomir Dependent MicroRNA-205 Reduction Enhances Adhesion Ability of Human Corneal Epithelial Keratinocytes |
| Jun Li1,2†, Hua Bai3†, Yong Zhu1, Xiao-yan Wang1, Fang Wang1, Jun-wu Zhang1, Robert M. Lavker4, and Jia Yu1* |
a Department of Biochemistry, Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing 100005, China
b Department of Cardiology, Chongqing First-aid Medical Center, Chongqing 400014, China
c Department of Ophthalmology, The Military General Hospital of Beijing PLA, Beijing 100700, China
d Department of Dermatology, Feinberg School of Medicine, Northwestern University, Chicago 60611, USA |
|
|
|
|
Abstract Objective To investigate the effect of microRNA-205 reduction by antagomirs on adhesion ability of normal human corneal epithelial keratinocytes (NHCEKs). Methods Antagomir-205, complementary and inhibitory to microRNA-205, was used to suppress endogenous microRNA-205 in NHCEKs. The adhesion ability of treated NHCEKs was then assessed by cell adhesion assay. Immunoblot and immunohistochemistry were conducted to determine the level of two focal adhesion-related proteins, focal adhesion kinase (FAK) and paxillin (Pax). Phalloidin staining was performed to measure the level of filamentous actin in antagomir-treated NHCEKs. Results Antagomir-205 markedly reduced the level of microRNA-205 in NHCEKs and significantly enhanced adhesion ability of NHCEKs (P<0.01). Further protein analysis validated that inhibition of microRNA-205 increased the number of phosphorylated FAK and phosphorylated Pax, and decreased filamentous actin. Conclusion Our findings suggest that microRNA-205 has down-regulating effect on cell motility in NHCEKs.
|
|
Received: 05 May 2010
Published: 10 June 2010
|
| Fund: Supported by Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences Grant (National Institutes of Health Grants (Corresponding author Tel: 86-10-65296423 |
| About author: Corresponding author Tel: 86-10-65296423, E-mail: j-yu@ibms.pumc.edu.cn |
| 1. |
Landgraf P, Rusu M, Sheridan R, et al. A mammalian mi-croRNA expression atlas based on small RNA library se-quencing. Cell 2007; 129:1401-14.
|
| 2. |
Lu L, Reinach PS, Kao WW. Corneal epithelial wound healing. Exp Biol Med 2001; 226:653-64.
|
| 3. |
Raja, Sivamani K, Garcia MS, et al. Wound re-epithelialization: modulating keratinocyte migration in wound healing. Front Biosci 2007; 12:2849-68.
|
| 4. |
Ma L, Teruya-Feldstein J, Weinberg RA. Tumour invasion and metastasis initiated by microRNA-10b in breast cancer. Nature 2007; 449:682-8.
|
| 5. |
Tavazoie SF, Alarcón C, Oskarsson T, et al. Endogenous human microRNAs that suppress breast cancer metastasis. Nature 2008; 451:147-52.
|
| 6. |
Valastyan S, Reinhardt F, Benaich N, et al. A pleiotropically acting microRNA, miR-31, inhibits breast cancer metastasis. Cell 2009; 137:1032-46.
|
| 7. |
Ryan DG, Oliveira-Fernandes M, Lavker RM. MicroRNAs of the mammalian eye display distinct and overlapping tissue specificity. Mol Vis 2006; 12:1175-84.
|
| 8. |
Yu J, Ryan DG, Getsios S, et al. MicroRNA-184 antagonizes microRNA-205 to maintain SHIP2 levels in epithelia. PNAS 2008; 105:19300-5.
|
| 9. |
Qian Y, Corum L, Meng Q, et al. PI3K induced actin filament remodeling through Akt and p70S6K1: implication of essential role in cell migration. Am J Physiol Cell Physiol 2004; 286:C153-63.
|
| 10 |
Dyson JM, O’Malley CJ, Becanovic J, et al. The SH2- containing inositol polyphosphate 5-phosphatase, SHIP-2, binds filamin and regulates submembraneous actin. J Cell Biol 2001; 155:1065-79.
|
| 11 |
Prasad NK, Decker SJ. SH2-containing 5’-inositol phos-phatase, SHIP2, regulates cytoskeleton organization and ligand-dependent down-regulation of the epidermal growth factor receptor. J Biol Chem 2005; 280:13129-36.
|
| 12 |
Vandermoere F, El Yazidi-Belkoura I, Demont Y, et al. Proteomics exploration reveals that actin is a signaling target of the kinase Akt. Mol Cell Proteomics 2007; 6: 114-24.
|
| 13 |
Leslie NR, Downes CP. PTEN function: how normal cells control it and tumour cells lose it. Biochem J 2004; 382:1-11.
|
| 14 |
Lim MA, Yang L, Zheng Y, et al. Roles of PDK-1 and PKN in regulating cell migration and cortical actin formation of PTEN-knockout cells. Oncogene 2004; 23:9348-58.
|
| 15 |
Salmena L, Carracedo A, Pandolfi PP. Tenets of PTEN tumor suppression. Cell 2008; 133:403-14.
|
| 16 |
Yurko MA, O’Toole EA, Woodley DT. Phosphorylation of focal adhesion kinase (pp125(FAK)) is increased in human keratinocytes induced to migrate by extracellular matrices. J Cell Physiol 2001; 188:24-32.
|
| 17 |
Trinkaus-Randall V, Kewalramani R, Payne J, et al. Calcium signaling induced by adhesion mediates protein tyrosine phosphorylation and is independent of pHi. J Cell Physiol 2000; 184:385-99.
|
| 18 |
Shi Q, Boettiger D. A novel mode for integrin-mediated signaling: tethering is required for phosphorylation of FAK Y397. Mol Biol Cell 2003; 14:4306-15.
|
| 19 |
Ridley AJ, Schwartz MA, Burridge K, et al. Cell migration: integrating signals from front to back. Science 2003; 302:1704-9.
|
| 20 |
Gregory PA, Bert AG, Paterson EL, et al. The miR-200 family and miR-205 regulate epithelial to mesenchymal transition by targeting ZEB1 and SIP1. Nat Cell Biol 2008; 10:593-601.
|
| 21 |
Wu H, Zhu S, Mo YY. Suppression of cell growth and invasion by miR-205 in breast cancer. Cell Res 2009; 19:439-48.
|
| 22 |
Iorio MV, Casalini P, Piovan C, et al. microRNA-205 regulates HER3 in human breast cancer. Cancer Res 2009; 69:2195-200.
|
| 23 |
Gandellini P, Folini M, Longoni N, et al. miR-205 Exerts tumor-suppressive functions in human prostate through down-regulation of protein kinase Cepsilon. Cancer Res 2009: 69:2287-95.
|
| 24 |
Song H, Bu G. MicroRNA-205 inhibits tumor cell migration through down-regulating the expression of the LDL re-ceptor-related protein 1. Biochem Biophys Res Commun 2009; 388:400-5.
|
| 25 |
Iorio MV, Ferracin M, Liu CG, et al. MicroRNA gene expression deregulation in human breast cancer. Cancer Res 2005; 65:7065-70.
|
| 26 |
Iorio MV, Visone R, Di Leva G, et al. MicroRNA signatures in human ovarian cancer. Cancer Res 2007; 67:8699-707.
|
| 27 |
Jiang J, Lee EJ, Gusev Y, et al. Real-time expression profiling of microRNA precursors in human cancer cell lines. Nucleic Acids Res 2005; 33:5394-403.
|
| 28 |
Tran N, McLean T, Zhang X, et al. MicroRNA expression profiles in head and neck cancer cell lines. Biochem Biophys Res Commun 2007; 358:12-7.
|
| 29 |
Lebanony D, Benjamin H, Gilad S, et al. Diagnostic assay based on hsa-miR-205 expression distinguishes squa- mous from nonsquamous non-small-cell lung carcinoma. J Clin Oncol 2009; 27:2030-7.
|
| 30 |
Aman MJ, Lamkin TD, Okada H, et al. The inositol phos-phatase SHIP inhibits Akt/PKB activation in B cells. J Biol Chem 1998; 273:33922-8.
|
| 31 |
Cantley LC. The phosphoinositide 3-kinase pathway. Science 2002; 296:1655-7.
|
| 32 |
Hawkins PT, Anderson KE, Davidson K, et al. Signaling through Class I PI3Ks in mammalian cells. Biochem Soc Trans 2006; 34:647-62.
|
| 33 |
Le Clainche C, Carlier MF. Regulation of actin assembly associated with protrusion and adhesion in cell migration. Physiol Rev 2008; 88:489-513.
|
| [1] |
Wen Zhang, Xuan Zhang, Xiao-li Wu, Liu-sheng He, Xiao-feng Zeng, Amrie C. Grammer and Peter E. Lipsky. Petition between TRAF2 and TRAF6 Regulates NF-κB Activation in Human B Lymphocytes[J]. Chinese Medical Sciences Journal, 2010, 25(1): 1
-12
. |
| [2] |
Xiao-xiao Liu and Yong-ping Jiang. Pharmacokinetic Study of a Novel Recombinant Human Granulocyte Colony-stimulating Factor in Rats[J]. Chinese Medical Sciences Journal, 2010, 25(1): 13
-19
. |
| [3] |
Feng-xia Wu, Li-jun Wu, Xiong-yan Luo, Ming-hui Yang, Zhong Tang, Chuan-mei Xie, Jing-guo Zhou, Jian-long Guan and Guo-hua Yuan. Expression of FLICE-inhibitory Protein in Synovial Tissue and Its Association with Synovial Inflammation in Juvenile Idiopathic Arthritis[J]. Chinese Medical Sciences Journal, 2010, 25(1): 20
-26
. |
| [4] |
Meng-tao Li, Jun Ai, Zhuang Tian, Quan Fang, Wen-jie Zheng and Xue-jun Zeng. Prevalence of Anti-endothelial Cell Antibodies in Patients with Pulmonary Arterial Hypertension Associated with Connective Tissue Diseases[J]. Chinese Medical Sciences Journal, 2010, 25(1): 27
-31
. |
| [5] |
Guang-wen Li, Guan-yi Zheng, Jin-guo Li and Xu-dong Sun. Relationship between Carotid Atherosclerosis and Cerebral Infarction[J]. Chinese Medical Sciences Journal, 2010, 25(1): 32
-37
. |
| [6] |
Song Fu, Zhi-cun Feng and Schranz Dietmar. Factors Influencing Pleural Effusion after Fontan Operation:an Analysis with 95 Patients[J]. Chinese Medical Sciences Journal, 2010, 25(1): 38
-43
. |
| [7] |
Jian Cao, Hui Zou, Bing-po Zhu, Hao Wang, Jian Li, Yu Ding and Xiao-ying Li. Sex Hormones and Androgen Receptor:Risk Factors of Coronary Heart Disease in Elderly Men[J]. Chinese Medical Sciences Journal, 2010, 25(1): 44
-49
. |
| [8] |
Yusoh Nuriyah, Xue-tao Ren, Li Jiang, Xi-pu Liu and Yan-hong Zou. Comparison between Ophthalmologists and Community Health Workers in Screening of Shallow Anterior Chamber with Oblique Flashlight Test[J]. Chinese Medical Sciences Journal, 2010, 25(1): 50
-52
. |
| [9] |
Hong-sheng Liu, Shan-qing Li, Ying-zhi Qin, Zhi-yong Zhang and Hua Ren. Surgical Treatment of Intralobar Pulmonary Sequestration[J]. Chinese Medical Sciences Journal, 2010, 25(1): 53
-56
. |
| [10] |
Ji-dong Wu, Yue-hong Zheng, Nim Choi and Furtado Rui. Revascularization for Iliac-femoral Artery Pseudoaneurysm with Greater Saphenous Vein[J]. Chinese Medical Sciences Journal, 2010, 25(1): 57
-60
. |
|
|
Viewed |
|
|
|
Full text
|
|
|
|
|
Abstract
|
|
|
|
|
Cited |
|
|
|
|
| |
Shared |
|
|
|
|
| |
Discussed |
|
|
|
|
2010, Vol. 25 
