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RNA干扰对胃癌中CD133(+)细胞生物学特性影响的研究

Study on the Biological Characteristics of CD133 (+) Cells Interfered by RNA Interference in Gastric Cancer.

作者信息

Yu Ji-Wei, Wang Shou-Lian, Wu Ju-Gang, Lu Rui-Qi, Ni Xiao-Chun, Cai Cheng, Jiang Bo-Jian

机构信息

Department of General Surgery, Shanghai 3rd People's Hospital, School of Medicine, Shanghai Jiao-Tong University, No. 280, Mohe Road, Shanghai 201900, China.

出版信息

ISRN Gastroenterol. 2014 Mar 19;2014:329519. doi: 10.1155/2014/329519. eCollection 2014.

DOI:10.1155/2014/329519
PMID:25006468
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC3977524/
Abstract

Background. To detect the changes of biological characteristics in gastric cancer cells interfered by CD133-specific small interfering RNA (siRNA). Methods. First to select the siRNA which has the strongest interference effect among 3 siRNAs (i.e., siRNA1, siRNA2, and siRNA3) in KATO-III cells by RT-PCR and Western blotting assays. Then, CD133(+) cells were sorted out from KATO-III cells using an immunomagnetic bead sorting method and transfected with the selected siRNA. Furthermore, the proliferating characteristics, the antichemotherapeutic assessment, Transwell invasion assay, monoclonal sphere formation assay, and subcutaneous transplanted tumor formation assay in nude mice were investigated. Results. siRNA3 showed the strongest interference effect in KATO-III cells. As compared to the uninterfered control group, the CD133(+) cells treated by siRNA3 showed significant decreases in the abilities of proliferation, invasion, clone sphere formation, and resistance to antitumour drugs as well as the weight and size of the transplanted tumor, which was nearly similar to that of CD133(-) cells. Additionally, the protein expression level of the EMT factor E-cadherin increased while those of EMT-related Snail and N-cadherin decreased in CD133(+) cells interfered by siRNA3. Conclusion. Inhibition of CD133 gene expression reduces the abilities of gastric cancer cells in proliferation, invasion, clonal sphere formation, and chemoresistance as well as tumor formation in nude mice.

摘要

背景。检测CD133特异性小干扰RNA（siRNA）干扰后胃癌细胞生物学特性的变化。方法。首先通过RT-PCR和蛋白质印迹分析在KATO-III细胞中从3种siRNA（即siRNA1、siRNA2和siRNA3）中筛选出干扰作用最强的siRNA。然后，采用免疫磁珠分选法从KATO-III细胞中筛选出CD133(+)细胞，并用筛选出的siRNA进行转染。此外，研究了裸鼠的增殖特性、抗化疗评估、Transwell侵袭实验、单克隆球形成实验和皮下移植瘤形成实验。结果。siRNA3在KATO-III细胞中显示出最强的干扰作用。与未干扰的对照组相比，用siRNA3处理的CD133(+)细胞在增殖、侵袭、克隆球形成和抗肿瘤药物耐药能力以及移植瘤的重量和大小方面均显著降低，这与CD133(-)细胞几乎相似。此外，在受siRNA3干扰的CD133(+)细胞中，EMT因子E-钙黏蛋白的蛋白表达水平升高，而EMT相关的Snail和N-钙黏蛋白的表达水平降低。结论。抑制CD133基因表达可降低胃癌细胞的增殖、侵袭、克隆球形成和化疗耐药能力以及裸鼠体内肿瘤的形成能力。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ad41/3977524/941fb5a65816/ISRN.GASTROENTEROLOGY2014-329519.009.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ad41/3977524/9b449aa53d20/ISRN.GASTROENTEROLOGY2014-329519.001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ad41/3977524/66ee3c093b76/ISRN.GASTROENTEROLOGY2014-329519.002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ad41/3977524/7f214ee39ecb/ISRN.GASTROENTEROLOGY2014-329519.003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ad41/3977524/f60d43696066/ISRN.GASTROENTEROLOGY2014-329519.004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ad41/3977524/99a207ed9a93/ISRN.GASTROENTEROLOGY2014-329519.005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ad41/3977524/333d8e360654/ISRN.GASTROENTEROLOGY2014-329519.006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ad41/3977524/668d9f692892/ISRN.GASTROENTEROLOGY2014-329519.007.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ad41/3977524/6b5cd37f793f/ISRN.GASTROENTEROLOGY2014-329519.008.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ad41/3977524/941fb5a65816/ISRN.GASTROENTEROLOGY2014-329519.009.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ad41/3977524/9b449aa53d20/ISRN.GASTROENTEROLOGY2014-329519.001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ad41/3977524/66ee3c093b76/ISRN.GASTROENTEROLOGY2014-329519.002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ad41/3977524/7f214ee39ecb/ISRN.GASTROENTEROLOGY2014-329519.003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ad41/3977524/f60d43696066/ISRN.GASTROENTEROLOGY2014-329519.004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ad41/3977524/99a207ed9a93/ISRN.GASTROENTEROLOGY2014-329519.005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ad41/3977524/333d8e360654/ISRN.GASTROENTEROLOGY2014-329519.006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ad41/3977524/668d9f692892/ISRN.GASTROENTEROLOGY2014-329519.007.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ad41/3977524/6b5cd37f793f/ISRN.GASTROENTEROLOGY2014-329519.008.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ad41/3977524/941fb5a65816/ISRN.GASTROENTEROLOGY2014-329519.009.jpg

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