Asian Pacific Journal of Cancer Prevention

Asian Pacific Journal of Cancer Prevention (아시아태평양암예방학회)
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Gene Silencing of β-catenin by RNAi Inhibits Proliferation of Human Esophageal Cancer Cells by Inducing G0/G1 Cell Cycle Arrest

  • Wang, Jin-Sheng (Central Laboratory, Peace Hospital Affiliated to Changzhi Medical University) ;
  • Ji, Ai-Fang (Central Laboratory, Peace Hospital Affiliated to Changzhi Medical University) ;
  • Wan, Hong-Jun (Jinci College, Shanxi Medical University) ;
  • Lu, Ya-Li (Department of Pathology and Pathophysiology, Basic School of Medicine, Jiamusi University) ;
  • Yang, Jian-Zhou (Central Laboratory, Peace Hospital Affiliated to Changzhi Medical University) ;
  • Ma, Li-Li (Central Laboratory, Peace Hospital Affiliated to Changzhi Medical University) ;
  • Wang, Yong-Jin (Central Laboratory, Peace Hospital Affiliated to Changzhi Medical University) ;
  • Wei, Wu (Central Laboratory, Peace Hospital Affiliated to Changzhi Medical University)
  • Published : 2012.06.30
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Abstract

Objectives: The aim of the present study was to explore mechanisms underlying the effects of down-regulating ${\beta}$-catenin expression on esophageal carcinoma (EC) cells. Methods: Cell cycle distribution and apoptosis were determined using flow cytometry and annexin V apoptosis assay, respectively. Transmission electron microscopy (TEM) was used to examine changes in ultrastructure, while expression of cyclin D1 protein and mRNA was detected by western blot and real-time PCR. Proliferating cell nuclear antigen (PCNA) and extracellular signal-regulated kinase (ERK) 1-2 were evaluated by Western blot analysis. PCNA labeling index (LI) was determined by immunocytochemistry. Results: Compared with pGen-3-con transfected and Eca-109 cells, the percentage of G0/G1-phase pGen-3-CTNNB1 transfected cells was obviously increased (P<0.05), with no significant difference among the three groups with regard to apoptosis (P>0.05). pGen-3-CTNNB1 transfected cells exhibited obvious decrease in cyclin D1 mRNA and protein expression (P<0.05) and the ultrastructure of Eca-109 cells underwent a significant change after being transfected with pGen-3-CTNNB1, suggesting that down-regulating ${\beta}$-catenin expression can promote the differentiation and maturation. The expression of PCNA and the ERKI/2 phosphorylation state were also down-regulated in pGen-3-CTNNB1 transfected cells (P<0.05). At the same time, the PCNA labeling index was decreased accordingly (P<0.05). Conclusion: Inhibition of EC Eca-109 cellproliferation by down-regulating ${\beta}$-catenin expression could improve cell ultrastructure by mediating blockade in G0/G1 through inhibiting cyclin D1, PCNA and the MAPK pathway (p-ERK1/2).

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References

  1. Coqueret O (2002). Linking cyclins to transcriptional control. Gene, 299, 35-55. https://doi.org/10.1016/S0378-1119(02)01055-7
  2. de Castro J, Gamallo C, Palacios J, et al (2000). Beta-catenin expression pattern in primary oesophageal squamous cell carcinoma. Relationship with clinicopathologic features and clinical outcome. Virchows Arch, 437, 599-604. https://doi.org/10.1007/s004280000266
  3. Deng YJ, Rong TH, Zhou J, et al (2007). Establishment of a human esophageal carcinoma transplantation model with MUC1 high expression in nude mice. Ai Zheng, 26, 693-7.
  4. Jemal A, Bray F, Center MM, et al (2011). Global cancer statistics. CA Cancer J Clin, 61, 69-90. https://doi.org/10.3322/caac.20107
  5. Ji L, CaoXF, Wang HM, et al (2007). Expression level of $\beta$-catenin is associated with prognosis of esophageal carcinoma. World J Gastroenterol, 13, 2622-5. https://doi.org/10.3748/wjg.v13.i18.2622
  6. Kimura Y, Shiozaki H, Doki Y, et al (1999). Cytoplasmic betacatenin in esophageal cancers. Int J Cancer, 84, 174-8. https://doi.org/10.1002/(SICI)1097-0215(19990420)84:2<174::AID-IJC14>3.0.CO;2-E
  7. Krishna M, Narang H (2008). The complexity of mitogenactivated protein kinases (MAPKs) made simple. Cell Mol Life Sci, 65, 3525-44. https://doi.org/10.1007/s00018-008-8170-7
  8. Livak KJ, Schmittgen TD (2001). Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) method. Methods, 25, 402-8. https://doi.org/10.1006/meth.2001.1262
  9. Meloche S, Pouyssegur J (2007). The ERK1⁄2 mitogen-activated protein kinase pathway as a master regulator of the G1-to S-phase transition. Oncogene, 26, 3227-39. https://doi.org/10.1038/sj.onc.1210414
  10. Nunez R (2001). DNA measurement and cell cycle analysis by flow cytometry. Curr Issues Mol Biol, 3, 67-70.
  11. Ren HZ, Wang JS, Pan GQ, et al (2010). Comparative proteomic analysis of $\beta$-catenin-mediated malignant progression of esophageal squamous cell carcinoma. Dis Esophagus, 23, 175-84. https://doi.org/10.1111/j.1442-2050.2009.01001.x
  12. Sears RC, Nevins JR (2002). Signaling networks that link cell proliferation and cell fate. J Biol Chem , 277, 11617-20. https://doi.org/10.1074/jbc.R100063200
  13. Shiina H, Igawa M, Shigeno K, et al (2002). Beta-catenin mutations correlate with over expression of C-myc and cyclin D1 genes in bladder cancer. J Urol, 168, 2220-6. https://doi.org/10.1016/S0022-5347(05)64359-5
  14. Shtutman M, Zhurinsky J, Simcha I, et al (1999). The cyclin D1 gene is a target of the beta-catenin/LEF-1 pathway. Proc Natl Acad Sci USA, 96, 5522-7. https://doi.org/10.1073/pnas.96.10.5522
  15. Takahashi Y, Kawate S, Watanabe M, et al (2007). Amplification of c-myc and cyclin D1 genes in primary and metastatic carcinomas of the liver. Pathol Int, 57, 437-42. https://doi.org/10.1111/j.1440-1827.2007.02120.x
  16. Tetsu O, McCormick F (1999). Beta-catenin regulates expression of cyclin D1 in colon carcinoma cells. Nature, 398, 422-6. https://doi.org/10.1038/18884
  17. Van Engeland M, Nieland LJ, Ramaekers FC, et al (1998). Annexin V-affinity assay: a review on an apoptosis detection system based on phosphatidylserine exposure. Cytometry, 31, 1-9. https://doi.org/10.1002/(SICI)1097-0320(19980101)31:1<1::AID-CYTO1>3.0.CO;2-R
  18. Veeramachaneni NK, Kubokura H, Lin L, et al (2004). Downregulation of beta-catenin inhibits the growth of esophageal carcinoma cells. J Thorac Cardiovasc Surg, 127, 92-8. https://doi.org/10.1016/j.jtcvs.2003.06.008
  19. Wang JS, Wei W, Ji AF, et al (2011). Abnormal expression of $\beta$-catenin in esophageal squamous cell carcinoma and its clinical significance. Zhong Guo Zu Zhi Hua Xue Yu Xi Bao Hua Xue Za Zhi, 20, 124-9.
  20. Wang JS, Ji AF, Wen JF, et al (2010). Effect of $\beta$-catenin gene silencing by shRNA on biologic characteristics of human esophageal carcinoma cells. Zhonghua Bing Li Xue Za Zhi, 39, 835-41.
  21. Wang JS, Zheng CL, Wang YJ, et al (2009). Gene silencing of $\beta$-catenin by RNAi inhibits cell proliferation in human esophageal cancer cells in vitro and in nude mice. Dis Esophagus, 22, 151-62. https://doi.org/10.1111/j.1442-2050.2008.00875.x
  22. Xu XL, Huang YJ, Wang YQ, et al(2011) . 2,3,4',5-Tetrahydroxystilbene-2-O-$\beta$- D-glucoside inhibits platelet-derived growth factor-induced proliferation of vascular smooth muscle cells by regulating the cell cycle. Clin Exp Pharmacol Physiol, 38, 307-13. https://doi.org/10.1111/j.1440-1681.2011.05502.x
  23. Zhou XB, Lu N, Zhang W, et al (2002). Expression and significance of beta-catenin in esophageal carcinoma. Ai Zheng, 21, 877-80.
  24. Zou XN, Taylor PR, Mark SD, et al (2002). Seasonal variation of food consumption and selected nutrient intake in Linxian, a high risk area for esophageal cancer in China. Int J Vitam Nutr Res, 72, 375-82. https://doi.org/10.1024/0300-9831.72.6.375

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