【摘要】 目的:检测纳米分子聚酰胺-胺(PAMAM)介导耐药基因-ATP结合转运蛋白G超家族成员2(ABCG2)沉默对宫颈癌侧群细胞的治疗效果。方法:从宫颈癌细胞中采用流式细胞术分离宫颈癌侧群细胞,检测ABCG2基因的表达,噻唑蓝(MTT)法检测多柔比星对细胞的抑制率;实验分为si RNA组、无义组和对照组,PAMAM介导ABCG2基因沉默后,蛋白质印迹法(western blot)检测ABCG2的表达,MTT法检测多柔比星对细胞敏感性的恢复情况;建立宫颈癌侧群细胞荷瘤鼠模型,给予多柔比星药物治疗,测量肿瘤体积。结果:宫颈癌侧群细胞ABCG2的蛋白水平高于非侧群细胞(P<0.05);MTT耐药实验显示,多柔比星对非侧群细胞的抑制率高于侧群细胞(P<0.01)。si RNA组在ABCG2表达沉默后,抑制率恢复到(69.4±11.3)%,与对照组及无义组比较差异均有统计学意义(P<0.01);动物实验结果显示,si RNA组肿瘤体积小于对照组和无义组(P<0.05)。结论:以PAMAM为介质可实现宫颈癌侧群细胞团中ABCG2基因表达沉默,恢复宫颈癌侧群细胞对化疗药物的治疗敏感性。更多还原

【Abstract】 Objective:To detect treatment effect on cervical cancer side population cells by nano-molecular polyamideamine(PAMAM)-mediated drug resistance gene ATP-binding cassette superfamily G member 2(ABCG2) silencing. Methods:The cervical cancer side population cells were isolated by flow cytometry assay from cervical cancer cells. The protein level of ABCG2 was detected by immunofluorescence method. MTT assay was used to detect the inhibition rate of tumor cells induced by doxorubicin. According to the experimental requirements, the three groups, si RNA group, antisense group and control group are set up. After the design and synthesis of si RNA targeting ABCG2 gene,it was transfected into tumor cells by PAMAM,then the ABCG2 gene expression was detected by western blot,and the restore of tumor cells to doxorubicin sensitivity by detected by MTT assay. Finally,these cells were transplated into the subcutaneous of de-thymus mice and the tumor volume was detected.Results:The protein level of ABCG2 in cervical cancer side population cells was higher than that of nonside population cells(P <0.05);The inhibition rate of doxorubicin to non side population cell was significantly higher than that of side population cells(P <0.01) by MTT,after ABCG2 gene silence,the inhibition rate in si RNA group recoverd to(69.4 ±11.3)%, showing significant differences with the control group and the non sense group(P<0.01). The animal experimental results show that the tumor volume in si RNA treatment group was significantly lower than the control group and antisense sequence group(P<<0.05).Conclusions:PAMAM can induce ABCG2 gene silencing in cervical cancer side population cell mass,to restore its treatment sensitivity to chemotherapeutic agents.更多还原