膜联蛋白A7低表达对胃癌MGC-803细胞生物学特性的影响

田焕娜 王小杰 陈龙 梁秀军 刘兰芳 李欣*

doi:10.16098/j.issn.0529-1356.2015.02.012

解剖学报 ›› 2015, Vol. 46 ›› Issue (2) : 216-220. DOI: 10.16098/j.issn.0529-1356.2015.02.012

肿瘤生物学

膜联蛋白A7低表达对胃癌MGC-803细胞生物学特性的影响

田焕娜¹王小杰¹陈龙¹梁秀军¹刘兰芳²李欣^1*

作者信息 +

Effect of annexin A7 knockdown on biological behaviour of MGC-803 cells

TIAN Huan-na¹ WANG Xiao-jie¹ CHEN Long¹ LIANG Xiu-jun¹LIU Lan-fang²LI Xin 1^*

Author information +

文章历史 +

摘要

目的探讨膜联蛋白A7(ANXA7)低表达对人胃癌低分化细胞MGC-803的影响。方法将细胞分为实验组、阴性对照组和空白对照组。采用RNA干扰技术将靶向ANXA7的siRNA和阴性对照siRNA以脂质体转染法分别转染胃癌细胞系MGC-803细胞，空白对照组不予任何处理。转染48h后，采用Western blotting 和实时定量-PCR法进行抑制效果的鉴定。检测ANXA7低表达对MGC-803细胞黏附、伸展、剥脱和迁移等生物学行为的影响。结果靶向ANXA7的siRNA可显著抑制ANXA7的表达；ANXA7表达受抑后，细胞黏附、伸展和剥脱能力降低，与阴性对照组和空白对照组相比，差异显著（P<0.05）。细胞迁移变化不显著。结论 ANXA7表达降低后MGC-803细胞行为学发生明显改变。

Abstract

Objective To explore the effect of annexin A7(ANXA7) knockdown on biological behaviour of MGC-803 cells. Methods The MGC-803 cells were grouped into experiment group, negative group and blank group. RNA interference technology was used to transfect the siRNA targeted ANXA7, scrambled siRNA into MGC-803 cells with lipofectine 2000 without any treatment to the blank group. After transfection for about 48 hours, Western blotting and Real-time PCR were used to assure the suppression of ANXA7 on both protein level and mRNA level. Cell adhesion assay, detachment assay, and spreading assay and wound healing assay were used to detect the adhesion, detachment, spreading and migration abilities of MGC-803 cells. Results The expression of ANXA7 was significantly suppressed in the cells which were transfected with ANXA7 siRNA. When ANXA7 expression was suppressed, the cell adhesion rate, detachment rate, spreading rate in experiment group were significantly lower than those in negative group and blank group(P<0.05). Compared with the negative group and blank group, migration ability of experiment group was not significantly different. Conclusion The biological behaviour of MGC-803 cells change obviously when ANXA7 expression is suppressed and ANXA7 could become potential target of therapeutic strategies.

导出引用

田焕娜王小杰陈龙梁秀军刘兰芳李欣*. 膜联蛋白A7低表达对胃癌MGC-803细胞生物学特性的影响[J]. 解剖学报. 2015, 46(2): 216-220 https://doi.org/10.16098/j.issn.0529-1356.2015.02.012

TIAN Huan-na WANG Xiao-jie CHEN Long LIANG Xiu-jun LIU Lan-fang LI Xin*. Effect of annexin A7 knockdown on biological behaviour of MGC-803 cells[J]. Acta Anatomica Sinica. 2015, 46(2): 216-220 https://doi.org/10.16098/j.issn.0529-1356.2015.02.012

参考文献

［1］Gerke V, Creutz CE, Moss SE. Annexins: linking Ca2+signaling to membrane dynamics［J］. Nat Rev Mol Cell Bio, 2005, 6(6):449-461.
［2］Rescher U, Gerke V. Annexinsunique membrane binding proteins with diverse functions［J］. J Cell Sci, 2004, 117(13):2631-2639.
［3］Fatimathas L, Moss SE. Annexins as disease modifiers［J］. Histol Histopathol, 2010, 25(4):527-532.
［4］Srivastava M, Bubendorf L, Srikantan V, et al. ANX7, a candidate tumor suppressor gene for prostate cancer［J］. Proc Natl Acad Sci USA, 2001, 98(8):4575-4580. 
［5］Srivastava M, Bubendorf L, Raffeld M, et al. Prognostic impact of ANX7-GTPase in metastatic and HER2-negative breast cancer patients［J］. Clin Cancer Res, 2004, 10(7):2344-2350.
［6］Kataoka TR, Ito A, Asada H, et al. Annexin VII as a novel marker for invasive phenotype of malignant melanoma［J］. Jpn J Cancer Res, 2000, 91(1):75-83.
［7］Yadav AK, Renfrow JJ, Scholtens DM, et al. Monosomy of chromosome 10 associated with dysregulation of epidermal growth factor signaling in glioblastomas［J］. JAMA, 2009, 302(3):276-289.
［8］Xi JM, Zhao Q. Expression of ANXA7 in gastric cancer tissues and their effects on the differentiation and metastasis of gastric cancer ［J］. Journal of Clinical and Experimental Medicin, 2010, 9(10):726-727. (in Chiense)
奚剑敏, 赵强. AnnexinA7表达与胃癌分化和转移关系研究［J］. 临床和实验医学杂志, 2010, 9(10):726-727.
［9］Creutz CE, Pazoles CJ, Pollard HB. Identification and purification of an adrenal medullary protein (synexin) that causes calcium-dependent aggregation of isolated chromaffin granules［J］. J Biol Chem, 1978, 253(8):2858-2866. 
［10］Srivastava M, Torosyan Y, Raffeld M, et al. ANXA7 expression represents hormonerelevant tumor suppression indifferent cancers［J］. Int J Cancer, 2007, 121(12): 2628-2636.
［11］Ji H, Moritz RL, Kim YS, et al. Analysis of Ras-induced oncogenic transformation of NIH-3T3 cells using differential display 2-DE proteomics ［J］. Electrophoresis, 2007, 28(12):1997-2008.
［12］Srivastava M, Bubendorf L, Nolan L, et al. ANX7 as a bio-marker in prostate and breast cancer progression［J］. Dis Markers, 2001, 17(2):115-120.
［13］Mussunoor S, Murray GI. The role of annexins in tumour development and progression［J］. J Pathol, 2008, 216(2): 131-140.
［14］Wang XJ, Li X. Influence of annexin A7 low expression on proliferation of HepG2 cells ［J］. Acta Anatomica Sinica, 2013,44(5) :656-660. (in Chiense)
王小杰, 李欣. 膜联蛋白A7低表达对人肝癌HepG2细胞增殖的影响［J］. 解剖学报, 2013, 44(5) :656-660.
［15］Wang ZQ, Tang JW, Wang ShQ, et al. Establishment of mouse hepatic cancer cell line Hca-F with knockdown of ANXA7 gene［J］. Academic Journal of Second Military Medical University, 2008, 29(9):1029-1033. (in Chiense)
王志强, 唐建武, 王绍青, 等. 膜联蛋白A7表达稳定下调的小鼠肝癌Hca-F细胞株的建立［J］. 第二军医大学学报, 2008, 29(9):1029-1033.
［16］Caterino M, Ruoppolo M, Orrù S, et al. Characterization of red cell membrane proteins as a function of red cell density: AnnexinⅦ in different forms of hereditary spherocytosis［J］. FEBS Lett, 2006, 580(28-29):6527-6532.
［17］Huang YH, Asma Saleem qazi, Mohmmed mibrahim, et al. Study of proliferation and invasion activity in different mouse hepatocarcinoma cell lines and location and isoform expression of ANXA7［J］. China Journal of Modern Medicine, 2012, 22(35):9-13. (in Chiense)
黄玉红, Asma saleem qazi, Mohmmed mibrahim, 等. 不同小鼠腹水型肝癌细胞增殖侵袭能力与膜联蛋白A7的亚细胞定位和亚型表达研究［J］. 中国现代医学杂志, 2012, 22(35):9-13.