The Primary Effect on the Proteome of ARID1A-mutated Ovarian Clear Cell Carcinoma is Downregulation of the Mevalonate Pathway at the Post-transcriptional Level.
The Primary Effect on the Proteome of ARID1A-mutated Ovarian Clear Cell Carcinoma is Downregulation of the Mevalonate Pathway at the Post-transcriptional Level.
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DOI：
10.1074/mcp.m116.062539
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发表时间：
2016-11
期刊：
影响因子：
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通讯作者：
Speicher DW
中科院分区：
文献类型：
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作者：
Goldman AR;Bitler BG;Schug Z;Conejo-Garcia JR;Zhang R;Speicher DW
ARID1A编码SWI/SNF染色质重塑复合体的一个亚单位，它的失活突变在超过一半的卵巢透明细胞癌病例中发现，更广泛地存在于大多数类型的癌症中。为了确定细胞内信号通路中ARID1A依赖的变化，我们对表达或不表达ARID1A的同基因卵巢透明细胞癌细胞系进行了蛋白质组分析。在含有野生型ARID1A的卵巢透明细胞癌细胞系OVCA429中，ARID1A基因的敲除主要导致甲氧戊酸途径的下调，甲戊酸途径是参与类异戊二烯合成、胆固醇合成和其他下游途径的重要代谢途径。在一项补充实验中，野生型ARID1A在含有突变ARID1A的卵巢透明细胞癌细胞系OVISE中的表达以交互方式影响甲氧戊酸途径。这些分析的一个引人注目的方面是，尽管只有5%的检测到的蛋白质组显示出显著的丰度变化，但甲氧戊酸途径中的大多数蛋白质都受到ARID1A状态的协调影响。当将蛋白质组学数据与我们之前发表的ARID1A在OVISE细胞系中异位表达的微阵列数据进行比较时，通常有相应的变化。然而，甲氧戊酸途径内的基因没有检测到依赖ARID1A的变化。这一差异表明甲氧戊酸途径不受ARID1A介导的转录直接调控，而可能是转录后调控。我们得出结论，ARID1A状态间接影响甲氧戊酸途径，并可能影响其他过程，包括糖原代谢和14-3-3介导的信号转导。此外，我们的发现表明，mRNA水平的变化有时不能很好地指示癌细胞中受基因操作影响的信号通路。
Inactivating mutations in ARID1A, which encodes a subunit of the SWI/SNF chromatin-remodeling complex, are found in over half of ovarian clear cell carcinoma cases and more broadly across most types of cancers. To identify ARID1A-dependent changes in intracellular signaling pathways, we performed proteome analyses of isogenic ovarian clear cell carcinoma cell lines with or without ARID1A expression. Knockout of ARID1A in an ovarian clear cell carcinoma cell line with wild-type ARID1A, OVCA429, primarily resulted in downregulation of the mevalonate pathway, an important metabolic pathway involved in isoprenoid synthesis, cholesterol synthesis, and other downstream pathways. In a complementary experiment, expression of wild-type ARID1A in an ovarian clear cell carcinoma cell line containing mutated ARID1A, OVISE, affected the mevalonate pathway in a reciprocal manner. A striking aspect of these analyses was that, although only 5% of the detected proteome showed significant abundance changes, most proteins in the mevalonate pathway were coordinately affected by ARID1A status. There were generally corresponding changes when comparing the proteomics data to our previously published microarray data for ectopic expression of ARID1A in the OVISE cell line. However, ARID1A-dependent changes were not detected for genes within the mevalonate pathway. This discrepancy suggests that the mevalonate pathway is not regulated directly by ARID1A-mediated transcription and may be regulated post-transcriptionally. We conclude that ARID1A status indirectly influences the mevalonate pathway and probably influences other processes including glycogen metabolism and 14-3-3-mediated signaling. Further, our findings demonstrate that changes in mRNA levels are sometimes poor indicators of signaling pathways affected by gene manipulations in cancer cells.