生物合成的MCT4siRNA抑制卵巢癌细胞的增殖作用

Bio-synthesized MCT4 siRNA inhibited proliferation of ovarian cancer cell

目的探索生物合成的单羧酸转运蛋白4(MCT4)siRNA对卵巢癌细胞的作用。方法将靶向单羧酸转运蛋白4(MCT4)的siRNA在大肠杆菌里进行合成,HPLC纯化后,转染卵巢癌细胞A2780和SKOV3,Western blot检测siRNA的干涉效果。利用葡萄糖、乳酸及氧消耗检测试剂盒检测细胞糖酵解水平,MTT实验检测siRNA对卵巢癌细胞增殖的抑制作用。结果利用大肠杆菌发酵获得mg级MCT4siRNA纯品。转染卵巢癌细胞A2780和SKOV3后Western blot显示能有效降低MCT4的表达,降低细胞的糖酵解水平,抑制卵巢癌细胞的增殖。结论生物合成的MCT4具有良好的生物活性,能显著降低靶蛋白的表达,发挥抑癌作用,具有发展成为新的核酸药物的潜力。

Objective To explore function of bio-synthesized MCT4siRNA in ovarian cancer cells.Methods MCT4siRNA was synthesized in E.coli and purified by HPLC.The purified siRNA was transfected in ovarian cell line A2780and SkOV3.Western blot was used to check MCT4protein interrupted by its siRNA.The glycolysis level was evaluated by assaying lactate level,glucose intake and oxygen consumption of cells.The proliferation of ovarian cells was examined by MTT assay kit.Results It obtained mg level purified MCT4siRNA from E.coli.Western blot showed that MCT4protein was decreased after transfected with siRNA.The glycolysis level was decreased and proliferation of ovarian cancer cells was inhibited by transfection with siRNA.Conclusion Bio-synthesized MCT4siRNA can significantly decreased MCT4expression and has good bio-activities in inhibiting function of MCT4in canccer cells.This research suggest bio-synthesized MCT4siRNA be recognized as new protential nuclear drug.