摘要
目的筛选含有Tip30的涎腺腺样囊性癌高转移细胞,检测TIP30蛋白在涎腺腺样囊性癌高转移细胞中的表达,检测携带外源性TIP30细胞的细胞周期分布变化。方法应用脂质体转染方法将TIP30导入ACCM细胞中,G418筛选阳性克隆,用免疫印记杂交法(Western blot)检测转染细胞中TIP30蛋白的表达;流式细胞仪测定细胞周期分布。结果筛选14天后,含有TIP30的ACC-M单克隆形成,TIP30蛋白在ACC-M中表达稳定;携带外源性TIP30的细胞G0～G1期比例增高,G2-M、S期细胞比例降低。结论ACC-M细胞能稳定表达外源基因TIP30,携带外源性TIP30的细胞G0-G1期细胞比例增高,G2～M、S期细胞比例降低,从而影响ACCM细胞的生长,是TIP30抑制涎腺腺样囊性癌细胞生长的可能机制之一。
Objective To screen the adenoid cystic carcinoma-M with TIP30 and to detect the expression of TIP30 and cell cycle distribution by flow cytometry in ACC - M cells line. Methods Wide type TIP30 gene was transfected into ACC - M by using lipofectamine. The positive cell clones with G418 were identified. Western blot detected the expression of TIP30 protein in the cells. Flow cytometry detected the cell cycle distribution. Results Fourteen days after the selection, G418 cell clones containing TIP30 were identified. Western blot revealed a steady expression of the TIP30 protein in the transfected cells. DNA flow cytometric analysis established an increased percentage of AAC-M with the TIP30 in G0-G1 phases. The cell percentage was decreased in the G2 ~ M,S phases. Conclusion ACC- M can steadily express exogene TIP30. An increased percentage of ACC-M cells with TIP30 might be a contributing factor of the TIP30 to inhibit the proliferation of ACC- M.
出处
《现代口腔医学杂志》
CAS
CSCD
北大核心
2005年第1期54-56,共3页
Journal of Modern Stomatology
