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目的:NDRG2是N-Myc downstream regulated gene家族的成员之一,与细胞增殖和分化相关。前期研究发现,抑制NDRG2表达可以提高宫颈癌Hela细胞对于顺铂的化疗敏感性,本研究采用RNA干扰技术抑制人宫颈癌Hela细胞中NDRG2基因的表达研究其对Hela细胞紫杉醇化疗敏感性的影响。方法:利用化学合成的针对NDRG2特异性siRNA瞬时转染Hela细胞株,采用RT-PCR和Western Blot检测NDRG2 mRNA和蛋白表达情况,通过MTT法检测其与对照细胞在顺铂作用下的体外存活率差异。SPSS11.0统计包处理,采用t检验,P<0.05为差异有统计学意义。结果:在mRNA和蛋白水平,化学合成的NDRG2特异性siRNA oligomer可使Hela细胞的NDRG2表达水平明显降低。在0.1、1、10、100、500、1000μg/mL紫杉醇浓度组,Negative-control和NDRG2siRNA细胞的相应细胞存活率分别为97.21±2.38、90.09±2.42、83.35±3.86、62.93±3.75、18.22±5.46、1.14±0.67和99.62±3.15、94.91±3.83、85.71±2.93、58.59±3.36、17.99±3.40、0.73±0.34,组间比较P值均大于0.05,差异无统计学意义。结论:抑制NDRG2表达不影响宫颈癌Hela细胞在紫杉醇作用下的细胞存活率,不能提高其对紫杉醇的化疗敏感性。进一步拓展了对该基因的功能认知。
Purpose: NDRG2 is a member of the N-Myc downstream regulated gene family and is involved in cell proliferation and differentiation. Previous studies found that inhibition of NDRG2 expression can improve the chemosensitivity of cervical cancer Hela cells to cisplatin. In this study, RNA interference technology was used to inhibit the expression of NDRG2 gene in human cervical cancer Hela cells to study its sensitivity to paclitaxel in Hela cells. METHODS: Hela cell lines were transiently transfected with NDRG2-specific siRNAs by chemical synthesis. The expression of NDRG2 mRNA and protein was detected by RT-PCR and Western Blot. The survival rates of NDRG2 and control cells in vitro were assayed by MTT assay difference. SPSS11.0 statistical package processing, using t test, P <0.05 for the difference was statistically significant. Results: At mRNA and protein levels, NDRG2-specific siRNA oligomer chemically synthesized significantly reduced NDRG2 expression in Hela cells. At 0.1, 1, 10, 100, 500 and 1000μg / mL paclitaxel concentration, the corresponding cell survival rates of Negative-control and NDRG2siRNA cells were 97.21 ± 2.38, 90.09 ± 2.42, 83.35 ± 3.86, 62.93 ± 3.75, 18.22 ± 5.46 , 1.14 ± 0.67 and 99.62 ± 3.15, 94.91 ± 3.83, 85.71 ± 2.93, 58.59 ± 3.36, 17.99 ± 3.40 and 0.73 ± 0.34 respectively. The P values in all groups were all higher than 0.05, with no significant difference. CONCLUSION: Inhibition of NDRG2 expression does not affect cell viability of Hela cells under paclitaxel treatment, and does not improve chemosensitivity to paclitaxel. Further expand the functional understanding of the gene.