Effects of annexin A5 knockdown on cell cycle of human gastric cancer MGC-803 and MKN-45 cells&nbsp;

WANG Xiao-jie ZHAO Yi-na LI Xin*

doi:10.16098/j.issn.0529-1356.2018.03.012

PDF(400 KB)

Acta Anatomica Sinica ›› 2018, Vol. 49 ›› Issue (3) : 342-346. DOI: 10.16098/j.issn.0529-1356.2018.03.012

Cancer Biology

Effects of annexin A5 knockdown on cell cycle of human gastric cancer MGC-803 and MKN-45 cells

WANG Xiao-jie ZHAO Yi-na LI Xin*

Author information +

History +

Abstract

Objective To investigate the effects of annexin A5 (ANXA5) konckdown on cell cycle of human gastric cancer cell lines MGC-803, MKN-45 cells. Methods Cells were diveded into interference group, negative control group and blank control group. The siRNA targeted to ANXA5 was transfected into MGC-803, MKN-45 cells by lipofectamine 2000 respectively. At the same time, negative siRNA was also transfected into negative control group and in blank control group nothing was added in. Real-time PCR and Western blotting was used to identify whether the siRNA targeted to ANXA5 can highly suppress annexin A5 expression 48 hours after transfection. When the effect of the siRNA was assured, Real-time PCR and Western blotting were applied to observe the expression of p21^cip1 mRNA and P21^cip1 protein, Western blotting was used to detect the expression of cyclinD1, flow cytometry(FCM) was applied to observe the cell cycle progression. Results The expression of p21^cip1 mRNA, P21^cip1 protein and cyclinD1 decreased significantly when compared with the negative control group and the blank control group(P<0.05). FCM analysis showed that the percentage of cells in G1 phase increased in interference group(P<0.05). Conclusion ANXA5 knockdown can down-regulate the expression of p21^cip1 mRNA, P21^cip1 protein and cyclinD1 in human gastric cancer MGC-803 and MKN-45 cells, it is speculated that ANXA5 may affect the development of gastric cancer by inducing cell cycle arrest by acting on cell cycle related proteins.

Key words

Annexin A5 / MGC-803 / MKN-45 / CyclinD1 / Real-time PCR / Western blotting / Flow cytometry / Human

Cite this article

EndNote

Ris (Procite)

Bibtex

Download Citations

WANG Xiao-jie ZHAO Yi-na LI Xin. Effects of annexin A5 knockdown on cell cycle of human gastric cancer MGC-803 and MKN-45 cells [J]. Acta Anatomica Sinica. 2018, 49(3): 342-346 https://doi.org/10.16098/j.issn.0529-1356.2018.03.012

References

［1］Gong L, Zhao H, Wang L, et al. Upregulation of annexin A5 affects the biological behaviors of lung squamous carcinoma cells in vitro［J］. Science Bulletin, 2014, 59(28):3610-3620.

［2］Peng B, Gao C, Guan H, et al. Annexin A5 as a potential marker in tumors［J］. Clin Chim Acta, 2014, 427:42-48.

［3］Wang B. Analyzing cell cycle checkpoints in response to ionizing radiation on Mammalian cells ［J］. Methods Mol Bio, 2014, 1170:313-320.

［4］Flores JM, Martin-Caballero J, Garcfa-Fernandez RA. P21 and p27 a shared senescence history［J］. Cell Cycle, 2014, 13(11):1655-1656.

［5］Chen A, Huang X, Xue Z, et al. The role of p21 in apoptosis, proliferation, cell cycle arrest, and antioxidant activity in UVB-irradiated human HaCaT keratinocytes［J］.Med Sci Monit Basic Res, 2015, 21:86-95.

［6］Piazuelo MB, Correa P. Gastric cancer: overview［J］. Colomb Med(Cali), 2013, 44(3):192-201.

［7］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.

［8］Wang XJ, Yang YL, Tian HN, et al. Effects of annexin A5 low expression on migration and invasion of HeLa cells［J］. Acta Anatomica Sinica, 2016, 47(4):487-492. (in Chinese)王小杰，杨艳丽，田焕娜，等. 膜联蛋白A5低表达对人宫颈癌HeLa细胞迁移和侵袭的影响［J］. 解剖学报, 2016, 47(4):487-492.

［9］Guo W, Man X, Yuan H, et al. Proteomic analysis on portal vein tumor thrombus associated proteins for hepatocellular carcinoma［J］. Natl Med J China, 2007, 87(30):2094-2097.

［10］Xue G, Hao L, Ding F, et al. Expression of annexin A5 is associated with higher tumor stage and poor prognosis in colorectal adenocarcinomas［J］. J Clin Gastroenterol, 2009, 43(9):831-837.

［11］Cui Y, Zhang D, Jia Q, et al. Proteomic and tissue array profiling identifies elevated hypoxia-regulated proteins in pancreatic ductal adenocarcinoma［J］. Cancer Invest, 2009, 27(7):747-755.

［12］Yong ST, Wang XF.A novel, non-apoptotic role for Scythe/BAT3: a functional switch between the pro-and anti-proliferative roles of p21 during the cell cycle［J］. PLoS One, 2012, 7(6):e38085.

［13］Wei M, Zhu L, Li Y, et al. Knocking down cyclin D1b inhibits breast cancer cell growth and suppresses tumor development in a breast cancer model［J］. Cancer Sci, 2011, 1202(8):1537-1544.

［14］Ru Y, Chen XJ, Zhao ZW, et al. CyclinD1 and p57kip2 as biomarkers in differentiation, metastasis and prognosis of gastric cardia adenocarcinoma［J］. Oncotarget, 2017, 8(43):73860-73870.

［15］Patel H, Abduljabbar R, Lai CF, et al. CDK7, cyclin H and MAT1 is elevated in breast cancer and is prognostic in estrogen receptor-positive breast cancer［J］. Clin Cancer Res, 2016, 22(3):5929-5938.