Department of Pathology, Xiangya Medical School, Central South University, Hunan Province, China.
Pathology. 2009;41(7):655-60. doi: 10.3109/00313020903273100.
Our current investigation attempts to study the role of the fascin1 gene in growth and metastasis of gastric cancer cell line MKN45.
Small interfering RNA (siRNA) was used to inhibit fascin1 expression in the human gastric cancer cell line MKN45. Expression of fascin1 in fascin1 siRNA transfected cells (sifascin1), non-transfected cells (NT) and non-specific fascin1 siRNA cells (CON) were examined by Western blotting and reverse transcription polymerase chain reaction (RT-PCR). Cell growth ability in vitro was evaluated by MTT and clone formation assays. Cell mobility in vitro was examined by the Boyden chamber assay. Nude mice metastasis models were established by abdominal cavity transfer method. Tumour growth was evaluated by immunohistochemistry with proliferating cell nuclear antigen (PCNA).
Knockdown of fascin1 expression in MKN45 cells resulted in decreased cellular proliferative and migratory abilities. In vitro, the cloning efficiency of siFascin1 cells (34.2%) was significantly lower compared to that in NT (78.5%) (p < 0.05). The migration rate in siFascin1 cells was significantly decreased (33.7%) compared with NT cells (89.4%) (p < 0.05). In vivo, the cell proliferation rate was lower in siFascin1 cells (25.8%) compared to that in NT (75.0%) (p < 0.05). The number of tumour clones in the liver was significantly lower in siFascin1 cells (2.0 +/- 1.1) compared to that in NT (5.1 +/- 1.6) (p < 0.05).
Our study demonstrates that down-regulation of fascin1 suppresses the proliferation and migration of gastric cancer cells MKN45, suggesting that fascin1 siRNA may offer a novel potential gene therapy approach for human gastric cancer with fascin1 over-expression.
本研究旨在探讨 fascin1 基因在人胃癌细胞系 MKN45 生长和转移中的作用。
应用小干扰 RNA（siRNA）抑制 fascin1 在人胃癌细胞系 MKN45 中的表达。Western blot 和逆转录聚合酶链反应（RT-PCR）检测 fascin1 siRNA 转染细胞（sifascin1）、未转染细胞（NT）和非特异性 fascin1 siRNA 细胞（CON）中 fascin1 的表达。MTT 法和克隆形成实验检测细胞体外生长能力，Boyden 室实验检测细胞体外迁移能力，腹腔转移法建立裸鼠转移模型，增殖细胞核抗原（PCNA）免疫组化检测肿瘤生长情况。
MKN45 细胞 fascin1 表达下调导致细胞增殖和迁移能力降低。体外实验中，siFascin1 细胞的克隆效率（34.2%）明显低于 NT 细胞（78.5%）（p<0.05）。siFascin1 细胞的迁移率（33.7%）明显低于 NT 细胞（89.4%）（p<0.05）。体内实验中，siFascin1 细胞的增殖率（25.8%）明显低于 NT 细胞（75.0%）（p<0.05）。siFascin1 细胞的肝内肿瘤克隆数（2.0±1.1）明显少于 NT 细胞（5.1±1.6）（p<0.05）。
本研究表明下调 fascin1 抑制胃癌细胞系 MKN45 的增殖和迁移，提示 fascin1 siRNA 可能为 fascin1 过表达的人类胃癌提供一种新的潜在基因治疗方法。
