抑制NDRG2基因表达对宫颈癌Hela细胞紫杉醇化疗敏感性的影响  

Regulatory Effects of RNA Interference against NDRG2 Gene on Chemosensitivity of Cervical Cancer Hela Cells to Paclitaxel

作　　者：杨乐[1,2] 张健[2] 张璟[2] 刘文超[3] 刘军叶[4,2] 

机构地区：[1]西安市中心医院肿瘤内科,陕西西安710003 [2]第四军医大学生物化学与分子生物学教研室肿瘤生物学国家重点实验室,陕西西安710032 [3]第四军医大学西京医院肿瘤中心,陕西西安710032 [4]第四军医大学放射医学教研室,陕西西安710032

出　　处：《现代生物医学进展》2013年第17期3230-3233,共4页Progress in Modern Biomedicine

基　　金：国家自然科学基金项目(81272490);西安市卫生局2011年度科研计划项目(J2011023)

摘　　要：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。Objective： NDRG2,a member of N-Myc downstream regulated gene family,plays some roles in cellular proliferation and differentiation.We have found that down-regulation of NDRG2 could enhance sensitivity of Hela cells to cisplatin.This study is to explore the effects of small interfering RNA（siRNA） specific to NDRG2 on chemosensitivity to paclitaxel of cervical cancer Hela cells.Methods： The siRNA specific for NDRG2 was synthesized and transfected into Hela cells by lipofectamine.NDRG2 mRNA expression and protein contents were detected by RT-PCR and Western Blot respectively.Under the action of chemotherapy drug paclitaxel,the changes in proliferation of Hela cells before and after the transfection of NDRG2 siRNA were studied by MTT.Statistical analyses were performed with the SPSS software（version 11.0）.t-test for independent groups.A P level of 0.05 was considered statistically significant.Results： Hela cells transfected with the NDRG2siRNA oligomer showed significantly reduced NDRG2 transcript and protein levels than cells transfected with negative-control oligomer.After treated with different concentration of paclitaxel,the growth inhibition rate of negative control and NDRG2siRNA 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±3.40,0.73±0.34 respectively,Statistical analysis showed on significant difference（P0.05）.Conclusion： Under the action of paclitaxel,down-regulation of NDRG2 can＇t influence the proliferation of Hela cells,neither does it increase sensitivity to paclitaxel in Cervical cancer Hela cells.It helps enrich the understanding of this gene＇s function.

关 键 词：NDRG2 RNA干涉 化疗敏感性 紫杉醇 HELA细胞 

分 类 号：R737.33[医药卫生—肿瘤]