Effects of annexin A5 low expression on migration and invasion of HeLa cells

doi:10.16098/j.issn.0529-1356.2016.04.009

Abstract

Abstract:

Objective To investigate the effects of annexin A5 (ANXA5) low expression on HeLa cell migration and invasion. Methods HeLa cells were divided into four groups, which were interference group, negative control group, and transfection reagent control group, and blank control group. RT-PCR and Western blotting were used to identify the inhibition efficiency at 72 hours after transfection. Cell scratch method and Transwell experiment were used to detect the ability of migration and invasion of HeLa cells in each group. E-cadherin and matrix metalloproteinase(MMP)-9 mRNA and protein were determined by RT-PCR and Western blotting respectively. Results HeLa cell ANXA5 expression on both mRNA and protein level was suppressed significantly(P<0.05). Compared with the negative control group, the abilities of migration and invasion of HeLa cells in the interference group were increased remarkably (P<0.05). The E-cadherin mRNA and protein expression of HeLa cells in interference group decreased while MMP-9 increased when compared with negative control group (P<0.05).
Conclusion The siRNA which targets ANXA5 can highly suppress ANXA5 expression. ANXA5 knockdown promotes migration and invasion of HeLa cells via upregulation of E-cadherin and downregulation of MMP-9.

Key words: Annexin A5, HeLa cell, Migration, Invasion, Cell wound scratch assay, Transwell assay

WANG Xiao-jie YANG Yan-jie2TIAN Huan-na LIAGN Xiu-jun CHEN Long LI Xin*. Effects of annexin A5 low expression on migration and invasion of HeLa cells[J]. AAS, 2016, 47(4): 487-492.

References

［1］Gerke V, Moss SE. Annexins: from structure to function［J］. Physiol Rev, 2002, 82(2):331-371.
［2］Li X, Gao FL, Li JT, et al. Expression of annexin A5 in human uterine cervical squamous cell carcinomas［J］. Acta Anatomica Sinica, 2008, 39(6):923926. (in Chinese)
李欣, 高福禄, 李建团, 等. 膜联蛋白A5在人子宫颈鳞癌组织中的表达［J］. 解剖学报, 2008, 39(6):923-926.
［3］Li X, Zhou XCh, Xu Q, et al. Influence of low expression of annexin A5 on the proliferation and apoptosis of HeLa cells［J］. Acta Anatomica Sinica, 2009, 40(1):48-51. (in Chinese)李欣, 周晓春, 许倩, 等. 膜联蛋白A5低表达对人宫颈癌细胞系HeLa细胞增殖与凋亡的影响［J］. 解剖学报, 2009, 40(1):48-51.
［4］Peng B, Guo C, Guan H, et al. Annexin A5 as a potential marker in tumors［J］. Clin Chim Acta, 2014, 427:42-48
［5］Ray ME, Mehra R, Sandler HM, et al. E-cadherin protein expression predicts prostate cancer salvage radiotherapy outcomes［J］. J Urol, 2006, 176(4 Pt1):1409-1414.
［6］Albelda SM. Role of integrins and other cell adhesion molecules in tumor progression and metastasis［J］. Lab Invest, 1993, 68(1):4-17.
［7］Nawrocki-Raby B, Gilles C, Polette M, et al. E-cadherin mediates MMP down-regulation in highly invasive bronchial tumor cells［J］. Am J Pathol, 2003, 162(2):653-661.
［8］Frixen UH, Behrens J, Sachs M, et al. E-cadherin-mediated cell-cell adhesion prevents invasiveness of human carcinoma cells ［J］. J Cell Biol, 1991, 113(1):173-185.
［9］Mareel M, Bracke M, Van Roy F, et al. Cancer metastasis: negative regulation by an invasion-suppressor complex［J］. Cancer Detect Prev, 1995, 19(5):451-464.
［10］Perl AK, Wilgenbus P, Dahl U, et al. A causal role for E-cadherin in the transition from adenoma to carcinoma［J］. Nature, 1998, 392(6672):190-193.
［11］Imao T, Koshida T, Endo Y, et al. Dominant role of E-cadherin in the progression of bladder cancer［J］. J Urol, 1999, 161(2):692-698.
［12］Lowy AM, Knight J, Groden J. Restoration of E-cadherin/β-catenin expression in pancreatic cancer cells inhibits growth by induction of apoptosis［J］. Surgery, 2002, 132(2) :141-148.
［13］Gong L, Zhao H, Wang L, et al. Upregulation of annexin A5 affects the biological behaviors of lung squamous carcinoma cells in vitro［J］. Oncology, 2014, 59(28):3610-3620.
［14］Lynch CC. Matrix metalloproteinases as master regulators of the vicious cycle of bone metastasis［J］. Bone, 2011, 48(1):44-53.
［15］Nelson AR, Fingleton B, Rothernerg ML, et al. Matrix metalloproteinases: biologic activity and clinical implications［J］. J Clin Oncol, 2000, 18(5):1135-1149.
［16］Yang LJ, Chen Zh. Expression of OPN and MMP-9 in colorectal cancer and the correlation with liver metastasis［J］. Contemporary Medicine, 2012, 18(12):7-9. (in Chinese)
杨丽君, 陈钟. OPN和MMP-9在结直肠癌中的表达及其与肝转移的相关性［J］. 当代医学, 2012, 18(12):7-9.
［17］Sun XH, Lü HB, Li H, et al.Expressions of MMP-9 and TIMP-1 mRNA in esophageal squamous cell carcinoma tissue and their significances［J］.Journal of Jilin University(Medicine Edition), 2013, 39(6):1247-1251. (in Chinese)
孙晓宏, 吕红博, 李卉, 等. MMP-9和TIMP-1mRNA在食管鳞状细胞癌组织中的表达及其意义［J］. 吉林大学学报(医学版), 2013, 39(6):1247-1251.

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