RKIP基因表达上调对宫颈癌细胞生物学行为影响

THE EFFECTS OF RKIP GENE UP-REGULATION EXPRESSION ON BIOLOGICAL BEHAVIOR OF CERVICAL CANCER CELLS

目的探讨Raf激酶抑制蛋白(RKIP)基因表达上调对宫颈癌细胞生物学行为的影响。方法应用脂质体法,将含有正义(ss)RKIP cDNA的真核表达载体转染入宫颈癌Caski细胞,G418筛选阳性克隆,Western-blot方法检测转染前后细胞RKIP的表达,建立RKIP基因表达上调的稳定转染细胞系(ssRKIP)。应用四甲基偶氮唑蓝(MTT)法、双层软琼脂集落形成实验、体外侵袭实验(Transwell小室)方法,观察RKIP基因对宫颈癌细胞生物学行为的影响。结果建立了RKIP表达上调的稳定转染宫颈癌Caski细胞系。RKIP基因转染后,与未转染细胞比较,对照空载体pcDNA3.1(+)转染细胞的RKIP蛋白表达无明显变化,含正义cDNA的ssRKIP细胞的RKIP蛋白表达水平则上调(t=8.137,P<0.01)。RKIP基因表达上调能够抑制宫颈癌Caski细胞的体外生长增殖、锚定非依赖性生长及体外侵袭能力。结论 RKIP基因对宫颈癌细胞的增殖、锚定非依赖性生长和侵袭等生物学行为均有抑制作用,其在宫颈癌发病中可能起重要作用。

Objective To study the effect of RKIP gene up-regulation on biological behavior of cervical cancer cells. Methods Using liposome technique, eukaryotic expression vector that contained （ss） RKIP cDNA was transfected into Caski cells of cervical cancer. G418 was used to select positive clones. The Western-blot was employed to detect the expression of RKIP, be- fore and after the transfection, a ssRKIP that up-regulated RKIP expression was created. The effect of RKIP on biological behavior of cervical cancer cells were observed by applying Methylthiazoletetrazolium （MTT）, soft agar colony formation and in vitro inva- sion method. Results Stably transfected Caski cell line that up-regulated RKIP expression was established. After transfection, compared with non-transfected ceils, the RKIP protein expression levels of control ceils transfected with empty vector pcDNA3.1 （＋） did not change significantly, the RKIP protein expression levels of ssRKIP cells increased （t= 8.137, P〈0.01）. Up-regulation of RKIP gene could inhibit the growth and proliferation of Caski cells in vitro, and anchorage-independent growth and invasion abi- lity. Conclusion The RKIP gene has an inhibitory action on the proliferation, anchorage-independent growth and invasion of cervical cancer cells, which probably plays an important role in the pathogenesis of this malignancy.