摘要
目的探究外源性膜联蛋白A5(ANXA5)对人胃癌MGC-803细胞增殖、凋亡和迁移的影响,并初步探讨其影响迁移的机制。方法制备ANXA5蛋白培养液,将人胃癌MGC-803细胞分为外源ANXA5组和空白对照组,分别用ANXA5蛋白培养液和等量培养基进行处理,培养24 h,倒置显微镜观察其形态学变化。MTT和Western blot法检测外源性ANXA5对细胞增殖的影响。PI荧光染色法对两组细胞的凋亡情况进行定量分析。细胞划痕实验检测两组细胞的迁移能力。Western blot法在蛋白水平上检测外源性ANXA5对E-钙黏蛋白(E-cadherin)和基质金属蛋白酶9(MMP-9)表达的影响。结果将ANXA5蛋白外源性作用于MGC-803细胞后,细胞发生了细胞变圆、胞浆呈空泡状等明显的形态学变化。外源ANXA5组MGC-803细胞增殖、迁移能力明显低于空白对照组(P<0.05),且外源ANXA5组凋亡细胞的数量明显增加(P<0.001)。外源ANXA5组E-cadherin的表达显著高于空白对照组(P<0.01),而MMP-9的表达低于空白对照组(P<0.05)。结论ANXA5蛋白可通过外源性添加的方式抑制人胃癌MGC-803细胞的增殖和迁移,并促进细胞凋亡,而且可能通过对E-cadherin和MMP-9的调控抑制了细胞的迁移。
Objective To explore the effects of exogenous annexin A5(ANXA5)on proliferation,apoptosis and migration of human gastric cancer MGC-803 cells,and to preliminarily discuss its mechanism.Methods ANXA5 protein culture solution was prepared,and human gastric cancer MGC-803 cells were divided into exogenous ANXA5 group and blank control group,which was treated with ANXA5 protein culture solution and an equal amount of culture medium for 24 h,respectively.Then the morphological changes were observed with an inverted microscope.The effects of exogenous ANXA5 on cell proliferation were detected by MTT and Western blot.PI fluorescence staining was used to quantitatively analyze the apoptosis in the two groups.The cell migration ability was detected by cell scratch assay.Western blot was used to detect the effect of exogenous ANXA5 on the expression of E-cadherin and matrix metalloproteinase 9(MMP-9)at the protein level.Results After MGC-803 cells were cultured with exogenous ANXA5 protein,the cells had obvious morphological changes such as cell rounding and vacuole-like cytoplasm.The proliferation and migration abilities of MGC-803 cells in exogenous ANXA5 group was significantly lower than those in blank control group(P<0.05),while the number of apoptotic cells in exogenous ANXA5 group was significantly higher(P<0.001).The expression of E-cadherin in exogenous ANXA5 group was significantly higher than that in control group(P<0.01),while MMP-9 expression was lower(P<0.05).Conclusion Exogenous ANXA5 protein can inhibit the proliferation and the migration of human gastric cancer MGC-803 cells,and promote the apoptosis.Exogenous ANXA5 protein may inhibit the cell migration by regulating E-cadherin and MMP-9.
作者
代雅蕊
张佳璐
王小杰
赵依纳
王欢
李欣
DAI Yarui;ZHANG Jialu;WANG Xiaojie;ZHAO Yina;WANG Huan;LI Xin(Basic Medical Institute,Chengde Medical University,Chengde 067000,China;Department of Reproductive Medicine,Hebei Maternity Hospital;Department of Histology and Embryology,Chengde Medical University)
出处
《山西医科大学学报》
CAS
2020年第11期1155-1160,共6页
Journal of Shanxi Medical University
基金
河北省高校省级重点学科项目(201304)
河北省高等学校科学技术研究重点项目(Z2014026,ZD2018058)
河北省科学技术研究与发展计划项目(10276142)
作者简介
代雅蕊,女,1997-05生,在读本科,E-mail:yaruidai@qq.com;通信作者:李欣,E-mail:hbcdlx@cdmc.edu.cn。
