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Src在胃癌腹腔转移多细胞团簇抗失巢凋亡中的作用
Role of Src protein in anoikis resistance of multicellular aggregates derived from peritoneal metastatic gastric cancer cell lines
in vitro - 第三军医大学学报 2017年39卷第22期 页码：2157-2162
- 作者机构：
400038 重庆，第三军医大学西南医院全军普通外科中心
- 作者简介：
[ 余佩武, E-mail: yupeiwu01@sina.com YU Peiwu, E-mail: yupeiwu01@sina.com
- 基金信息：国家自然科学基金面上项目(81272366);重庆市基础科学与前沿技术研究(CSTC 2016jcyjA0104)
DOI：10.16016/j.1000-5404.201709157
中图分类号： R394.2;R730.23;R735.2收稿：2017-09-21，
修回：2017-10-17，
纸质出版：2017-11-30
- 稿件说明：
移动端阅览
何晨晨, 唐波, 董慧, 等. Src在胃癌腹腔转移多细胞团簇抗失巢凋亡中的作用[J]. 第三军医大学学报, 2017,39(22):2157-2162.
Chenchen HE, Bo TANG, Hui DONG, et al. Role of Src protein in anoikis resistance of multicellular aggregates derived from peritoneal metastatic gastric cancer cell lines
in vitro [J]. Journal of Third Military Medical University, 2017, 39(22): 2157-2162.何晨晨, 唐波, 董慧, 等. Src在胃癌腹腔转移多细胞团簇抗失巢凋亡中的作用[J]. 第三军医大学学报, 2017,39(22):2157-2162. DOI： 10.16016/j.1000-5404.201709157.
Chenchen HE, Bo TANG, Hui DONG, et al. Role of Src protein in anoikis resistance of multicellular aggregates derived from peritoneal metastatic gastric cancer cell lines
in vitro [J]. Journal of Third Military Medical University, 2017, 39(22): 2157-2162. DOI： 10.16016/j.1000-5404.201709157.
摘要
目的
2
探讨Src蛋白对体外三维培养的胃癌多细胞团簇(multicellular aggregates
MCAs)增殖及凋亡的影响。
方法
2
体外三维培养人胃腺癌细胞BGC823和MKN45成MCAs，Western blot检测MCAs和单层贴壁培养的胃癌细胞中Src蛋白表达情况。siRNA法干扰Src表达，转染胃癌细胞后检测其对胃癌细胞BGC823和MKN45 MCAs形态学的影响。分别采用CCK-8和流式细胞术检测干扰Src表达对胃癌BGC823和MKN45 MCAs细胞活力和细胞凋亡率的影响。
结果
2
利用BGC823和MKN5成功建立胃癌MCAs体外三维培养模型，Western blot检测结果表明，与对照组比较，MCAs中Src的表达水平明显高于单层贴壁培养的胃癌细胞[BGC823：(0.615±0.068)
vs
(0.219±0.017)，
P
<
0.01；MKN45：(0.657±0.087)
vs
(0.420±0.015)，
P
<
0.05
]
。干扰胃癌细胞BGC823和MKN45 Src表达，可以显著降低MCAs形成能力，并明显影响MCAs活力。流式细胞术检测结果表明，干扰Src表达可以显著诱导MCAs发生凋亡[BGC823：(9.456±0.209)
vs
(3.026±0.201)，
P
<
0.01；MKN45：(9.356±0.416)
vs
(2.541±0.251)，
P
<
0.05
]
。
结论
2
Src是胃癌MCAs抗失巢凋亡的关键分子，抑制Src表达可以显著抑制胃癌BGC823 MCAs和MKN45 MCAs形成，影响胃癌MCAs增殖，并诱导胃癌MCAs凋亡。
Abstract
Objective
2
To investigate the role of Src in the proliferation and apoptosis of multicellular aggregates (MCAs) derived from peritoneal metastatic gastric cancer cell lines in a three-dimensional (3D) culture system.
Methods
2
Human gastric adenocarcinoma cell lines BGC823 and MKN45 were induced to form MCAs in a 3D culture system. The expression of Src protein in the MCAs and in dispersed intraperitoneal free cancer cells was detected using Western blotting. A lentiviral siRNA vector expressing human Src was transfected in the cells and the morphological changes of BGC823 and MKN45 MCAs were observed; the viability and apoptosis of the MCAs were assessed using CCK-8 assay and flow cytometry
respectively.
Results
2
We successfully established a 3D culture system to simulate MCA formation by native gastric cancer cells in the peritoneal cavity. The results of Western blotting showed significantly increased expression levels of Src protein in the MCAs derived from BGC823 and MKN45 cells as compared with their monolayers in routine culture (0.615±0.068
vs
0.219±0.017
P
<
0.01; 0.657±0.087
vs
0.420±0.015
P
<
0.05). Inhibiting Src expression in BGC823 and MKN45 cells with the lentiviral siRNA vector significantly suppressed the ability of the cells to form MCAs and obviously lowered the viability of the MCAs. The siRNA-mediated Src knockdown effectively induced apoptosis of the MCAs derived from BGC823 (9.456±0.209
vs
3.026±0.201
P
<
0.01) and MKN45 cells (9.356±0.416
vs
2.541±0.251
P
<
0.05).
Conclusion
2
Src is a key molecule that plays a vital role in anoikis resistance of peritoneal metastatic gastric cancer cell clusters. Inhibition of Src expression significantly suppresses the formation of MCAs in BGC823 and MKN45 cells and induces proliferation suppression and apoptosis of the MCAs.
关键词
Keywords
references
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