Guo Jiao, Xu Shaohang, Huang Xuanlin, Li Lin, Zhang Congmin, Pan Qingfei, Ren Zhen, Zhou Ruo, Ren Yan, Zi Jin, Wu Lin, Stenvang Jan, Brünner Nils, Wen Bo, Liu Siqi
CAS Key Laboratory of Genome Sciences and Information, Beijing Institute of Genomics, Chinese Academy of Sciences , Beijing, 100101, China.
University of Chinese Academy of Sciences, Beijing, 100049, China.
J Proteome Res. 2016 Nov 4;15(11):4047-4059. doi: 10.1021/acs.jproteome.6b00387. Epub 2016 Aug 16.
A priority in solving the problem of drug resistance is to understand the molecular mechanism of how a drug induces the resistance response within cells. Because many cancer cells exhibit chromosome aneuploidy, we explored whether changes of aneuploidy status result in drug resistance. Two typical colorectal cancer cells, HCT116 and LoVo, were cultured with the chemotherapeutic drugs irinotecan (SN38) or oxaliplatin (QxPt), and the non- and drug-resistant cell lines were selected. Whole exome sequencing (WES) was employed to evaluate the aneuploidy status of these cells, and RNAseq and LC-MS/MS were implemented to examine gene expression at both mRNA and protein level. The data of gene expression was well-matched with the genomic conclusion that HCT116 was a near diploid cell, whereas LoVo was an aneuploid cell with the increased abundance of mRNA and protein for these genes located at chromosomes 5, 7, 12, and 15. By comparing the genomic, transcriptomic, and proteomic data, the LoVo cells with SN38 tolerance showed an increased genome copy in chromosome 14, and the expression levels of the genes on this chromosome were also significantly increased. Thus, we first observed that SN38 could impact the aneuploidy status in cancer cells, which was partially associated with the acquired drug resistance.
解决耐药性问题的一个优先事项是了解药物如何在细胞内诱导耐药反应的分子机制。由于许多癌细胞表现出染色体非整倍性，我们探究了非整倍性状态的变化是否会导致耐药性。将两种典型的结肠癌细胞HCT116和LoVo用化疗药物伊立替康（SN38）或奥沙利铂（QxPt）进行培养，并筛选出非耐药和耐药细胞系。采用全外显子组测序（WES）来评估这些细胞的非整倍性状态，并实施RNA测序和液相色谱-串联质谱（LC-MS/MS）以检测mRNA和蛋白质水平的基因表达。基因表达数据与基因组结论高度匹配，即HCT116是近二倍体细胞，而LoVo是具有位于5号、7号、12号和15号染色体上这些基因的mRNA和蛋白质丰度增加的非整倍体细胞。通过比较基因组、转录组和蛋白质组数据，对SN38耐受的LoVo细胞在14号染色体上显示出基因组拷贝增加，并且该染色体上基因的表达水平也显著增加。因此，我们首次观察到SN38可影响癌细胞的非整倍性状态，这与获得性耐药部分相关。
