Secretion of Mutant DNA and mRNA by the Exosomes of Breast Cancer Cells

被引:18
作者
Andreeva, Olga E. [1 ]
Shchegolev, Yuri Y. [1 ]
Scherbakov, Alexander M. [1 ]
Mikhaevich, Ekaterina I. [1 ]
Sorokin, Danila V. [1 ]
Gudkova, Margarita V. [1 ]
Bure, Irina V. [2 ]
Kuznetsova, Ekaterina B. [2 ]
Mikhaylenko, Dmitry S. [2 ]
Nemtsova, Marina V. [2 ]
Bagrov, Dmitry V. [3 ]
Krasil'nikov, Mikhail A. [1 ]
机构
[1] Minist Hlth Russia, NN Blokhin Natl Med Res Ctr Oncol, Inst Carcinogenesis, Dept Expt Tumor Biol, Moscow 115522, Russia
[2] Sechenov Univ, IM Sechenov First Moscow State Med Univ, Inst Mol Med, Lab Med Genet, Moscow 119991, Russia
[3] Lomonosov Moscow State Univ, Fac Biol, Dept Bioengn, Moscow 119234, Russia
基金
俄罗斯科学基金会; 俄罗斯基础研究基金会;
关键词
exosomes; breast cancer cells; DNA transferring; cancer; signalling; STRANDED GENOMIC DNA; EXTRACELLULAR VESICLES; SERUM EXOSOMES; KRAS;
D O I
10.3390/molecules26092499
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Exosomes are the small vesicles that are secreted by different types of normal and tumour cells and can incorporate and transfer their cargo to the recipient cells. The main goal of the present work was to study the tumour exosomes' ability to accumulate the parent mutant DNA or RNA transcripts with their following transfer to the surrounding cells. The experiments were performed on the MCF7 breast cancer cells that are characterized by the unique coding mutation in the PIK3CA gene. Using two independent methods, Sanger sequencing and allele-specific real-time PCR, we revealed the presence of the fragments of the mutant DNA and RNA transcripts in the exosomes secreted by the MCF7 cells. Furthermore, we demonstrated the MCF7 exosomes' ability to incorporate into the heterologous MDA-MB-231 breast cancer cells supporting the possible transferring of the exosomal cargo into the recipient cells. Sanger sequencing of the DNA from MDA-MB-231 cells (originally bearing a wild type of PIK3CA) treated with MCF7 exosomes showed no detectable amount of mutant DNA or RNA; however, using allele-specific real-time PCR, we revealed a minor signal from amplification of a mutant allele, showing a slight increase of mutant DNA in the exosome-treated MDA-MB-231 cells. The results demonstrate the exosome-mediated secretion of the fragments of mutant DNA and mRNA by the cancer cells and the exosomes' ability to transfer their cargo into the heterologous cells.
引用
收藏
页数:7
相关论文
共 19 条
[1]   Tumour microvesicles contain retrotransposon elements and amplified oncogene sequences [J].
Balaj, Leonora ;
Lessard, Ryan ;
Dai, Lixin ;
Cho, Yoon-Jae ;
Pomeroy, Scott L. ;
Breakefield, Xandra O. ;
Skog, Johan .
NATURE COMMUNICATIONS, 2011, 2
[2]   SRY gene transferred by extracellular vesicles accelerates atherosclerosis by promotion of leucocyte adherence to endothelial cells [J].
Cai, Jin ;
Guan, Weiwei ;
Tan, Xiaorong ;
Chen, Caiyu ;
Li, Liangpeng ;
Wang, Na ;
Zou, Xue ;
Zhou, Faying ;
Wang, Jialiang ;
Pei, Fang ;
Chen, Xinjian ;
Luo, Hao ;
Wang, Xinquan ;
He, Duofen ;
Zhou, Lin ;
Jose, Pedro A. ;
Zeng, Chunyu .
CLINICAL SCIENCE, 2015, 129 (03) :259-269
[3]   Extracellular vesicle-mediated transfer of donor genomic DNA to recipient cells is a novel mechanism for genetic influence between cells [J].
Cai, Jin ;
Han, Yu ;
Ren, Hongmei ;
Chen, Caiyu ;
He, Duofen ;
Zhou, Lin ;
Eisner, Gilbert M. ;
Asico, Laureano D. ;
Jose, Pedro A. ;
Zeng, Chunyu .
JOURNAL OF MOLECULAR CELL BIOLOGY, 2013, 5 (04) :227-238
[4]   Exosomes: key players in cancer and potential therapeutic strategy [J].
Dai, Jie ;
Su, Yangzhou ;
Zhong, Suye ;
Cong, Li ;
Liu, Bang ;
Yang, Junjun ;
Tao, Yongguang ;
He, Zuping ;
Chen, Chao ;
Jiang, Yiqun .
SIGNAL TRANSDUCTION AND TARGETED THERAPY, 2020, 5 (01)
[5]   Micronuclei as biomarkers of DNA damage, aneuploidy, inducers of chromosomal hypermutation and as sources of pro-in fl ammatory DNA in humans [J].
Fenech, Michael ;
Knasmueller, Siegfried ;
Bolognesi, Claudia ;
Holland, Nina ;
Bonassi, Stefano ;
Kirsch-Volders, Micheline .
MUTATION RESEARCH-REVIEWS IN MUTATION RESEARCH, 2020, 786
[6]   Indication of Horizontal DNA Gene Transfer by Extracellular Vesicles [J].
Fischer, Stefanie ;
Cornils, Kerstin ;
Speiseder, Thomas ;
Badbaran, Anita ;
Reimer, Rudolph ;
Indenbirken, Daniela ;
Grundhoff, Adam ;
Brunswig-Spickenheier, Baerbel ;
Alawi, Malik ;
Lange, Claudia .
PLOS ONE, 2016, 11 (09)
[7]   KRAS and BRAF mutations in serum exosomes from patients with colorectal cancer in a Chinese population [J].
Hao, Yi-Xin ;
Li, Yong-Mei ;
Ye, Ming ;
Guo, Yan-Yan ;
Li, Qiu-Wen ;
Peng, Xiu-Mei ;
Wang, Qi ;
Zhang, Shu-Fang ;
Zhao, Hui-Xia ;
Zhang, He ;
Li, Guang-Hui ;
Zhu, Jian-Hua ;
Xiao, Wen-Hua .
ONCOLOGY LETTERS, 2017, 13 (05) :3608-3616
[8]   Identification of Double-stranded Genomic DNA Spanning All Chromosomes with Mutated KRAS and p53 DNA in the Serum Exosomes of Patients with Pancreatic Cancer [J].
Kahlert, Christoph ;
Melo, Sonia A. ;
Protopopov, Alexei ;
Tang, Jiabin ;
Seth, Sahil ;
Koch, Moritz ;
Zhang, Jianhua ;
Weitz, Juergen ;
Chin, Lynda ;
Futreal, Andrew ;
Kalluri, Raghu .
JOURNAL OF BIOLOGICAL CHEMISTRY, 2014, 289 (07) :3869-3875
[9]   Discovery of Double-Stranded Genomic DNA in Circulating Exosomes [J].
Kalluri, Raghu ;
LeBleu, Valerie S. .
TARGETING CANCER, VOL 81, 2016, 2016, 81 :275-280
[10]   Enhanced shedding of extracellular vesicles from amoeboid prostate cancer cells Potential effects on the tumor microenvironment [J].
Kim, Jayoung ;
Morley, Samantha ;
Minh Le ;
Bedoret, Denis ;
Umetsu, Dale T. ;
Di Vizio, Dolores ;
Freeman, Michael R. .
CANCER BIOLOGY & THERAPY, 2014, 15 (04) :409-418