KISS-1 抑制胃癌细胞的增殖和侵袭。
KISS-1 inhibits the proliferation and invasion of gastric carcinoma cells.
机构信息
Department of Pathology, Xinxiang Medical University, Xinxiang 453003, Henan Province, China.
出版信息
World J Gastroenterol. 2012 Apr 21;18(15):1827-33. doi: 10.3748/wjg.v18.i15.1827.
AIM
To investigate the function of the KISS-1 gene in gastric carcinoma cells and to explore its potential mechanism.
METHODS
A KISS-1 eukaryotic expression vector was constructed and transfected into BGC-823 cells. Resistant clones were obtained through G418 selection. reverse transcription-polymerase chain reaction and western blotting were used to detect KISS-1 and matrix metalloproteinase-9 (MMP-9) expression in transfected cells. The growth of transfected cells was investigated by 3-(4, 5-dimethylthiazolyl-2)-2,5-diphenyltetrazolium bromide (MTT) proliferation assays, and the cells' invasive potential was analyzed by basement membrane (Matrigel) invasion assays. The anti-tumor effects of KISS-1 were tested in vivo using allografts in nude mice.
RESULTS
The expression level of KISS-1 mRNA and protein in BGC-823/KISS-1 transfected cells were significantly higher than in BGC-823/pcDNA3.1 transfected cells (P < 0.05) or the parental BGC-823 cell line (P < 0.05). The expression level of MMP-9 mRNA and protein in BGC-823/KISS-1 were significantly less than in BGC-823/pcDNA3.1 (P < 0.05) or BGC-823 cells (P < 0.05). MTT growth assays show the proliferation of BGC-823/KISS-1 cells at 48 h (0.642 ± 0.130) and 72 h (0.530 ± 0.164) were significantly reduced compared to BGC-823/pcDNA3.1 (0.750 ± 0.163, 0.645 ± 0.140) (P < 0.05) and BGC-823 cells (0.782 ± 0.137, 0.685 ± 0.111) (P < 0.05). Invasion assays indicate the invasive potential of BGC-823/KISS-1 cells (16.50 ± 14.88) is significantly reduced compared to BGC-823/pcDNA3.1 (20.22 ± 14.87) (P < 0.05) and BGC-823 cells after 24 h (22.12 ± 16.12) (P < 0.05). In vivo studies demonstrate the rate of pcDNA3.1-KISS-1 tumor growth is significantly slower than pcDNA3.1 and control cell tumor growth in nude mice. Furthermore, tumor volume of pcDNA3.1-KISS-1 tumors (939.38 ± 82.08 mm(3)) was significantly less than pcDNA3.1 (1250.46 ± 44.36 mm(3)) and control tumors (1284.36 ± 55.26 mm(3)) (P < 0.05). Moreover, the tumor mass of pcDNA3.1-KISS-1 tumors (0.494 ± 0.84 g) was significantly less than pcDNA3.1 (0.668 ± 0.55 g) and control tumors (0.682 ± 0.38 g) (P < 0.05).
CONCLUSION
KISS-1 may inhibit the proliferation and invasion of gastric carcinoma cells in vitro and in vivo through the downregulation of MMP-9.
目的
研究 Kiss-1 基因在胃癌细胞中的作用，并探讨其潜在机制。
方法
构建 Kiss-1 真核表达载体并转染 BGC-823 细胞，通过 G418 筛选获得抗性克隆。采用逆转录-聚合酶链反应和 Western blot 检测转染细胞中 Kiss-1 和基质金属蛋白酶-9（MMP-9）的表达。通过 3-(4,5-二甲基噻唑-2)-2,5-二苯基四唑溴盐（MTT）增殖实验研究转染细胞的生长情况，通过基底膜（Matrigel）侵袭实验分析细胞的侵袭能力。在裸鼠同种异体移植模型中检测 Kiss-1 的抗肿瘤作用。
结果
BGC-823/KISS-1 转染细胞中 Kiss-1 mRNA 和蛋白的表达水平明显高于 BGC-823/pcDNA3.1 转染细胞（P<0.05）或亲本 BGC-823 细胞系（P<0.05）。BGC-823/KISS-1 中 MMP-9 mRNA 和蛋白的表达水平明显低于 BGC-823/pcDNA3.1（P<0.05）或 BGC-823 细胞（P<0.05）。MTT 生长实验显示 BGC-823/KISS-1 细胞在 48 h（0.642±0.130）和 72 h（0.530±0.164）的增殖明显低于 BGC-823/pcDNA3.1（0.750±0.163，0.645±0.140）（P<0.05）和 BGC-823 细胞（0.782±0.137，0.685±0.111）（P<0.05）。侵袭实验表明 BGC-823/KISS-1 细胞的侵袭潜力（16.50±14.88）明显低于 BGC-823/pcDNA3.1（20.22±14.87）（P<0.05）和 BGC-823 细胞（22.12±16.12）（P<0.05）。体内研究表明，pcDNA3.1-KISS-1 肿瘤的生长速度明显慢于 pcDNA3.1 和对照组肿瘤在裸鼠中的生长速度。此外，pcDNA3.1-KISS-1 肿瘤的体积（939.38±82.08 mm3）明显小于 pcDNA3.1（1250.46±44.36 mm3）和对照组肿瘤（1284.36±55.26 mm3）（P<0.05）。此外，pcDNA3.1-KISS-1 肿瘤的瘤质量（0.494±0.84 g）明显小于 pcDNA3.1（0.668±0.55 g）和对照组肿瘤（0.682±0.38 g）（P<0.05）。
结论
Kiss-1 可能通过下调 MMP-9 来抑制胃癌细胞在体外和体内的增殖和侵袭。
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