脆性组氨酸三联体基因FHIT对乳腺癌细胞株MCF-7的作用

目的 克隆人类脆性组氨酸三联体基因(FHIT)并构建其真核表达载体pcDNA3.1(+)/FHIT,将人FHIT基因转染人乳腺细胞MCF-7中稳定表达,检测转染后细胞生物学特性的变化.方法 构建FHIT基因表达载体pcDNA3.1(+)/FHIT,用脂质体法将FHIT基因的真核质粒表达载体pcDNA3.1(+)/FHIT导入人乳腺癌细胞MCF-7中,细胞计数、流式细胞术分析转染后细胞的生物学特性变化.结果 将逆转录-聚合酶链反应(RT-PCR)扩增后的产物克隆到表达载体peDNA 3.1(+)上,经PCR扩增筛选鉴定和测序鉴定证实为所需序列,转染乳腺癌MCF-7细胞后,FHIT基因的表达明显增强,细胞周期分析发现MCF-7/FHIT与MCF-7比较S期及G2/M期的细胞减少,G0/G1期的细胞增加,MCF-7/FHIT细胞的凋亡率为(29.75±5.90)%,MCF-7细胞的凋亡率为(11.21±6.10)%,和MCF-7比较,转染后的MCF-7/FHIT细胞的凋亡明显增加,MCF-7细胞的增殖明显抑制.结论 FHIT基因表达载体可有效抑制乳腺癌细胞MCF-7的增殖.

Transfection of FHTT into human MCF-7 cell line and its biological characteristics

Objective To construct FHIT eukaryotic expression vector and transfect FHIR gene into human MCF-7 breast carcinoma cell line.and analyze the effects of FHIT on MCF-7.Methods FHIT eukaryotic expression vector pcDNA3.1(+)/FHIT Was constructed.FHIT full length cDNA was transfected into MCF-7 cell line by the vector of pcDNA3.1(+)/FHIT,and the transfectant with stable expression of FHIT was obtained by clone selection.The effects of FHIT on breast carcinoma cells were analyzed.including the alteration of MCF-7 cell lines in cell cycle.apoptosis and rate of growth.Results pcDNA3.1 (+)/FHIT was successfully constructed.and the expression of FHIT in MCF-7 cells inhibited the growth of breast carcinoma cells.Compared with MCF-7 cells.MCF-7/FHIT cells had significandy decreased number in S and G2/M-phases.and increased number in G0/G1-phase.The apoptosis rate of MCF-7/FHIT and MCF-7 Was(29.75±5.90)%,and(11.21±6.10)% resepctively.As compared with MCF-7 cells.the apoptotic MCF-7/FHIT cells were significantly increased.Conclusion Transfection of FHIT inhibited growth and proliferation,and induce apoptosis of breast carcinoma cells MCF-7.