ABCG2-SiRNA转染前后Hep-2细胞系侧群细胞含量变化的研究

Content change of sp cell subsets in hep-2 cell lines before and after ABCG2-SiRNA transfection

目的探讨腺苷三磷酸结合区转运蛋白G超家族成员-2(ABCG2)基因失活前后hep-2细胞系中侧群细胞含量的差别。方法体外培养hep-2细胞,应用荧光染料Hoechst33342让对数期hep-2细胞进行染色,对照组加入荧光染料Hoechst33342及MDRI(多重耐药基因multiple drug resistance)抑制剂维拉帕米,流式细胞仪检测SP细胞亚群含量;ABCG2-SiRNA转染hep-2细胞系后,应用荧光染料Hoechst33342对转染后细胞进行染色,对照组加入荧光染料Hoechst33342及维拉帕米,流式细胞仪检测SP细胞亚群含量,比较转染前后SP细胞亚群含量变化。结果 ABCG2-SiRNA转染前hep-2细胞Hoechst33342组SP细胞含量(3.15±0.32)%,Hoechst33342+维拉帕米组SP细胞含量(0.4±0.11)%;ABCG2-SiRNA转染后hep-2细胞Ho-echst33342组SP细胞含量(1.15±0.22)%,Hoechst33342+维拉帕米组SP细胞含量(0.0±0.09)%,转染后SP细胞亚群含量明显降低(P<0.05)。结论 ABCG2基因在喉癌SP细胞的发生发展过程中起重要作用,可能成为喉癌靶向治疗的潜在靶点。

Objective To investgate the contents of side population cells in hep-2 cell lines before and after the inactivation of ABCG2 gene. Methods Hep-2 cells were cultured in vitro and fluorescent dye of Hoechst33342 was utilized to stain hep-2 cells, for the other group, the fluorescent dye of Hoechst33342 ＋ Verapamil shall be added, flow cytometry was used to detect the content difference of sp cell subsets;ABCG2-SiRNA was transfected into Hep-2 cell lines, and utilize fluorescent dye of Hoechst33342 to stain the transfected cells. The control group shall add fluorescent dye of Hoechst33342 ＋ Verapamil, and flow cytometry was used to detect the content of sp cell subsets, and compare the contents of sp cell subsets before and after transfection. Results Before the transfection of ABCG2-SiR- NA, the content of sp cells in Hoechst33342 group of hep-2 cells was（3.15 ± 0.32）%, in the group of Hoechst33342 ＋ Verapamil, the content of sp cells was （ 0.4 ± 0.11 ） % ; After the transfection of ABCG2-SiRNA, the content of sp cells in Hoeehst33342 group of hep-2 cells was（ 1.15 ＋ 0.22）%, in the group of Hoeehst33342 ＋ Verapamil ,the content of sp ceils was （0.0 ± 0.09 ） %. Conclusion ABCG2 gene plays an important role in the occurrence and devel- opment of laryngeal cancer SP cells and may become a potential target of cancer therapy.