Bloom解旋酶基因RNA干扰载体对前列腺癌PC3细胞的抑制作用

发布时间：2018-06-03 21:14

本文选题：RNA干扰 + 前列腺肿瘤　；参考：《第二军医大学学报》2016年06期

【摘要】：目的采用RNA干扰(RNAi)技术抑制前列腺癌PC3细胞系中Bloom解旋酶基因的表达,探讨Bloom解旋酶基因表达下调后对PC3细胞的抑制作用。方法使用实验室前期成功构建的两条针对于Bloom解旋酶基因的RNAi载体shRNA-1和shRNA-2转染前列腺癌PC3细胞,以未转染RNAi载体为对照组,分别在转染24、48、72h后通过MTT法检测细胞增殖情况,转染48h后通过Transwell小室法检验细胞侵袭、迁移能力,细胞划痕实验检测细胞迁移情况,Hoechst/PI双染法检测细胞凋亡情况。结果转染RNAi载体后,与未转染对照组相比,各实验时间点的转染组细胞增殖率降低(P0.05),Transwell细胞侵袭和迁移实验中穿过室膜的细胞数与对照组比均减少(侵袭:119±24、118±30vs 227±38;迁移:122±13、121±47vs 277±32,P0.05),划痕愈合率与划痕迁移距离均降低,48h时差异有统计学意义(P0.05),而且细胞凋亡明显增加。结论以RNAi载体干扰Bloom解旋酶基因的表达可抑制前列腺癌PC3细胞的增殖、迁移和侵袭能力并促进其凋亡,为前列腺癌的靶向基因治疗提供了依据。
[Abstract]:Objective to inhibit the expression of Bloom helicase gene in prostate cancer PC3 cell line by RNA interference RNAi technique and to investigate the inhibitory effect of Bloom helicase gene expression on PC3 cells. Methods two RNAi vectors shRNA-1 and shRNA-2 targeting the Bloom helicase gene were used to transfect the prostate cancer PC3 cells. The untransfected RNAi vector was used as the control group. The proliferation of the cells was detected by MTT assay 72 hours after transfection. After 48 hours of transfection, cell invasion and migration were detected by Transwell chamber assay, cell migration was detected by scratch assay and apoptosis was detected by Hoechst/ Pi double staining. Results after transfection of RNAi vector, compared with the control group, The number of cells passing through the ventricular membrane in the invasion and migration of P0.05 30vs cells in each experimental time point decreased compared with that in the control group (invasion: 119 卤24118 卤30vs 227 卤38; migration: 122 卤13121 卤47vs 277 卤32P 0.05). The rate of scratch healing and the distance between scratch migration and scratch healing were decreased at 48h. The difference was statistically significant (P 0.05), and apoptosis increased significantly. Conclusion interfering the expression of Bloom helicase gene with RNAi vector can inhibit the proliferation, migration, invasion and apoptosis of prostate cancer PC3 cells, and provide the basis for the targeted gene therapy of prostate cancer.
【作者单位】：贵州大学生命科学学院;贵州大学动物科学学院高原山地动物遗传育种与繁殖教育部重点实验室;贵州大学农学院;
【基金】：国家自然科学基金(31361406) 贵州省国际合作项目[黔科合外G字(2011)7008号]~~
【分类号】：R737.25

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