TIP30蛋白在ACC-M细胞中的表达鉴定及作用

目的 :筛选含有TIP3 0的涎腺腺样囊性癌高转移细胞 ,检测TIP3 0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TIP3 0细胞的细胞周期分布变化。方法 :应用脂质体转染方法将TIP3 0导入ACC M细胞中 ,G418筛选阳性克隆 ,用免疫印记杂交法 (Westernblot)检测转染细胞中TIP3 0蛋白的表达 ;流式细胞仪测定细胞周期分布。结果 :筛选 14d后 ,含有TIP3 0的ACC M克隆形成 ,TIP3 0蛋白在ACC M中表达稳定 ;携带外源性TIP3 0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TIP3 0 ,携带外源性TIP3 0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ACC M细胞的生长 ,是TIP3 0抑制涎腺腺样囊性癌细胞生长的可能机制之一。

The effect of TIP30 on cell cycle distribution of ACC-M cells

Objective: To study the effect of TIP30 gene on the cell cy cle distribution in adenoid cystic carcinoma ACC-M cells. Methods: Wild type TIP30 gene was transfected into ACC-M cells by using lipofectamin e, the postive cell clone was selected with G418,the expression of TIP30 protei n in the cells was detected by Western blot. Cell cycle distribution was detecte d by flow cytometry. Results:14 days after seletion anti-G418 c ell clone was obtained. Wstern blot revealed that the TIP30 protein was expresse d in the transfected cells.Flow cytometric analysis indicated that TIP30 induced a arrest of the cells in G 0-G 1 phase and led to a decrease of cell percent age in G 2-M and S phase. Conclusion:ACC-M can stably express exogene TIP30.TIP30 can inhibit proliferation of ACC-M cells.