shrna干扰蛋白激酶d—2基因提高tca8113对化疗药物敏感性探究

《shrna干扰蛋白激酶d—2基因提高tca8113对化疗药物敏感性探究》由会员上传分享，免费在线阅读，更多相关内容在学术论文-天天文库。

1、shRNA干扰蛋白激酶D—2基因提高Tca8113对化疗药物敏感性探究　　[摘要]目的探讨蛋白激酶D（PKD）-2基因沉默对Tca8113细胞增殖、细胞程序性死亡及对化疗药物敏感性的影响。方法构建针对pkd-2基因的shRNA干扰质粒及空载体对照组质粒，建立pkd-2基因沉默的稳定细胞株；采用甲基噻唑基四唑（MTT）方法检测shRNA干扰后细胞株的增殖情况及对化疗药物的半数抑制质量浓度（IC50）；采用流式细胞术检测pkd-2基因沉默前后细胞程序性死亡率及对化疗药物的敏感性；采用免疫组化方法检测pkd-2沉默后细胞中P糖蛋白（P-gp）的表达。结果筛选并建立pkd-2基因沉默

2、的稳定细胞株；经shRNA干扰后Tca8113细胞的增殖速度与野生型细胞相比差异无统计学意义，但IC50明显降低；pkd-2沉默后Tca8113细胞程序性死亡率明显上升，且对化疗药物的敏感性显著提高；与野生型对照组Tca8113相比较，shRNA-pkd-2基因沉默的Tca8113经化疗药物诱导后P-gp表达明显下降。结论shRNA干扰技术特异性沉默Tca8113中的pkd-2基因，促进了肿瘤细胞的程序性死亡，降低对化疗药物的IC50，并明显提高肿瘤细胞对化疗药物的敏感性，同时降低了P-gp的表达。[关键词]蛋白激酶D；shRNA干扰；细胞程序性死亡12[中图分类号]Q78[

3、文献标志码]A[doi]10.7518/hxkq.2014.01.019TheshRNA-mediateddownregulationofproteinkinaseD-2enhancedchemosensitivityofTca8113DaiLina1，ZhangPing2，SuQin2.（1.Dept.ofChildren’sDentalCentral，TheAffiliatedHospitalofStomatology，ChongqingMedicalUniver-sity，ChongqingKeyLaboratoryforOralDiseasesandBiomedica

4、lSciences，Chongqing400015，China；2.StateKeyLaboratoryofOralDiseases，WestChinaHospitalofStomatology，SichuanUniversity，Chengdu610041，China）[Abstract]ObjectiveToexploretheeffectofsilencingproteinkinaseD（PKD）-2onTca8113cellproliferation，pro-grammedcelldeath，andchemosensitivity.MethodsThestablece

5、lllinesofpkd-2genesilencingandemptyvectorplasmidgroupwereestablished.Theproliferationand50%inhibitoryconcentration（IC50）ofshRNA-mediatedTca8113chemotherapydrugsweredetectedthroughmethylthiazolyltetrazoliumassay（MTT）.The12programmedcelldeathrateandsensitivitytoTca8113chemotherapydrugsbeforeo

6、rafterpkd-2genesilencingweremeasuredthroughflowcytometry.P-glycoprotein（P-gp）expressionofpkd-2silencingcellswasidentifiedbyimmunohistochemicalmethods.ResultsStablecelllinesofpkd-2genesilencingwereestablished.Comparedwithparentalcells，theproliferationofshRNA-mediatedTca8113wasnotsignificantl

7、ydifferent，butitsIC50waslower.Meanwhile，cellprogrammeddeathrateandsensitivitytochemotherapeuticdrugsofshRNA-mediatedTca8113significantlyincreased.ComparedwithwildgroupTca8113，asignificantdecreaseinP-gpex-pressionwasinducedbychemotherapydrugswithshRNA-pkd