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敲减BLM解旋酶表达增强前列腺癌PC3细胞对丝裂霉素C的敏感性
引用本文:吴萍,许厚强,赵佳福,刘忠伟,段志强,陈福,谌颖莲. 敲减BLM解旋酶表达增强前列腺癌PC3细胞对丝裂霉素C的敏感性[J]. 中国生物化学与分子生物学报, 2017, 33(2): 190-197. DOI: 10.13865/j.cnki.cjbmb.2017.02.13
作者姓名:吴萍  许厚强  赵佳福  刘忠伟  段志强  陈福  谌颖莲
基金项目:国家自然科学基金(No.31361406)资助
摘    要:丝裂霉素C是一种广谱抗肿瘤抗生素,对多种癌症有抗癌作用,其作用原理可使细胞的DNA发生链间交联,引起DNA双链断裂,阻碍DNA的复制,从而抑制肿瘤细胞分裂。临床上主要用于胃癌、肠癌、肝癌及胰腺癌等消化道癌方面的治疗。本文研究丝裂霉素C对转染人BLM解旋酶基因(shRNA载体)前后前列腺癌PC3细胞活性的影响。使用前期成功构建的干扰载体转染PC3细胞,在转染48 h后加药,通过荧光定量PCR、MTT法、Transwell小室实验、细胞划痕实验、流式细胞术,分别检测加药12、24、36 h BLM基因的表达量、PC3细胞增殖能力、侵袭能力、迁移能力及凋亡情况的变化。结果显示,敲减BLM基因表达后的PC3细胞相对于正常PC3细胞其增殖能力、侵袭能力和迁移能力能显著被丝裂霉素C抑制,且丝裂霉素C能显著促进其细胞的凋亡,说明BLM基因低表达的前列腺癌细胞对丝裂霉素C更敏感。研究结果为丝裂霉素C在前列腺癌的临床治疗上奠定了理论基础。

关 键 词:丝裂霉素C  前列腺癌细胞  BLM解旋酶  shRNA载体  细胞活性  
收稿时间:2016-11-23

Knockdown of BLM Helicase Promotes the Sensitivity of Prostate Cancer PC3 Cells to Mitomycin C
WU Ping,XU Hou Qiang,ZHAO Jia-Fu,LIU Zhong-Wei,DUAN Zhi-Qiang,CHEN FU,CHEN Ying-Lian. Knockdown of BLM Helicase Promotes the Sensitivity of Prostate Cancer PC3 Cells to Mitomycin C[J]. Chinese Journal of Biochemistry and Molecular Biology, 2017, 33(2): 190-197. DOI: 10.13865/j.cnki.cjbmb.2017.02.13
Authors:WU Ping  XU Hou Qiang  ZHAO Jia-Fu  LIU Zhong-Wei  DUAN Zhi-Qiang  CHEN FU  CHEN Ying-Lian
Abstract:Mitomycin C, a broad spectrum anticancer drug, can induce inter-strand cross-link and double-stranded breaks, thereby blocking DNA replication. Clinically, it is used in the treatment of gastrointestinal cancers such as gastric cancer, colorectal cancer, hepatoma and pancreatic cancer. This study aimed to analyze the effects of mitomycin C on the proliferation, migration and invasion of prostate cancer PC3 cells transfected with human BLM helicase shRNA plasmid. After 48 hours transfection, the cells were treated with mitomycin C for 12, 24 and 36 hours followed by qRT-PCR, MTT, Transwell chamber, wound scratch and flow cytometry assays. The levels of BLM helicase and the proliferation, migration, invasion and apoptosis of PC3 cells transfected with BLM helicase shRNA were analyzed. The results show that the proliferation, migration and invasion of BLM helicase-deficient PC3 cells was significantly inhibited by mitomycin C when compared to the control PC3 cells, whereas the apoptosis in BLM helicase-deficient PC3 cells was significantly promoted. The results indicate that the BLM helicase-deficient PC3 cells had higher sensitivity to mitomycin C, which suggests that mitomycin C may be applied for the clinic treatment of prostate cancer.
Keywords:mitomycin C   prostate cancer cells   BLM helicase   RNAi vector   cell viability  
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