ABCC1促进人正常肺上皮细胞BEAS-2B恶性转化的作用研究

Study on the role of ABCC1 in promoting malignant transformation of normal human lung epithelial cell line BEAS-2B

目的探讨ABCC1对人正常肺上皮细胞BEAS-2B细胞增殖、侵袭和迁移等恶性表型的影响。方法在人正常肺上皮细胞BEAS-2B细胞中,通过慢病毒途径过表达ABCC1,RT-PCR和Western Blot分别检测ABCC1 mRNA及蛋白的表达;CCK-8和流式细胞周期检测ABCC1对BEAS-2B细胞增殖的影响;Transwell检测ABCC1对BEAS-2B细胞侵袭能力的影响;细胞划痕实验检测ABCC1对BEAS-2B细胞迁移能力的影响。结果重组病毒感染BEAS-2B细胞后72h,RT-PCR和Western Blot结果显示Lv-ABCC1组ABCC1基因的mRNA和蛋白表达较Lv-Control组与对照组明显增强,说明成功建立了ABCC1高表达BEAS-2B细胞;流式细胞周期分析表明Lv-ABCC1组细胞较对照组细胞进入DNA合成期(S期)的比例明显增加;Transwell检测结果显示Lv-ABCC1组细胞侵袭能力明显增强;划线法检测显示Lv-ABCC1组细胞迁移能力明显增强。结论ABCC1能够显著增加BEAS-2B细胞的增殖、侵袭和迁移能力,促进BEAS-2B细胞的恶性转化。

Objective To explore the effect of ABCC1 on the proliferation,invasion and migration of normal human lung epithelial cells line BEAS-2B.Methods The overexpression of ABCC1 in BEAS-2B cell line was established by lentivirus.Real-time quantitative RT-PCR and Western Blot were used to detect the expression of ABCC1 mRNA and protein,respectively.CCK-8 and flow cytometry were used to detected the cell proliferation.Transwell assay was used to detected the effect of ABCC1 on the invasion ability of BEAS-2B cell line,and the cell scratch test was used to detect the effect of ABCC1 on the migration ability of BEAS-2B cell line.Results 72 hours after the recombinant virus infection,the results showed that the mRNA and protein expressions of ABCC1 in the Lv-ABCC1 group was significantly increased compared with the Lv-Control group and the control group.Flow cytometry analysis showed that the cells of the synthesis phase in the Lv-ABCC1 group increased significantly compared with the control group.Transwell assay results showed that the cell invasion activity of the Lv-ABCC1 group was significantly enhanced,and the cell scratch detection showed that the cell migration ability of the Lv-ABCC1 group was also significantly enhanced.Conclusion ABCC1 can significantly increase the proliferation,invasion and migration capacity of BEAS-2B cell line,and promote the malignant transformation of BEAS-2B cells.