The roles of mutated SPINK1 gene in prostate cancer cells
The roles of mutated SPINK1 gene in prostate cancer cells
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突变的SPINK1基因在前列腺癌细胞中的作用
DOI：
10.1093/mutage/geac019
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发表时间：
2022-09-16
期刊：
影响因子：
2.7
通讯作者：
Chen，Ni
中科院分区：
文献类型：
--
作者：
Pan，Xiuyi;Tan，Junya;Chen，Ni
SPINK1阳性前列腺癌(PCa)是一种侵袭性前列腺癌亚型。然而，除了来源于人前列腺癌CWR22R异种移植瘤的22Rv1细胞外，目前还缺乏明确的研究来阐明SPINK1在大多数前列腺癌细胞中表达缺失的潜在机制。本研究旨在探讨SPINK1蛋白阳性/阴性表达的机制及其在PCa细胞中的生物学作用。与LNCaP、C4-2B、DU145和PC-3细胞相比，22Rv1细胞高表达SPINK1mRNA，且仅在22Rv1细胞中检测到该蛋白。在这些细胞系中，野生型SPINK1编码序列仅在22Rv1细胞中发现，在其他细胞系中发现了两个突变位点：c.194G>；A错义突变和c.210T>；C同义突变。我们的进一步研究表明，这些突变与SPINK1mRNA和蛋白水平的降低有关。功能实验表明，SPINK1促进PC-3细胞的增殖、迁移和侵袭，而敲除SPINK1则抑制22Rv1细胞的增殖、迁移和侵袭。野生型SPINK1基因比突变的SPINK1基因更能促进细胞的恶性行为。流式细胞仪细胞周期分析显示，SPINK1降低G0/G1期细胞百分率，增加S期细胞百分率。我们发现SPINK1基因的c.194G>；A和c.210T>；C突变降低了该基因的mRNA和蛋白水平。野生型SPINK1基因与PCa细胞的侵袭性生物学行为有关，可能成为治疗PCa的潜在靶点。
SPINK1-positive prostate cancer (PCa) has been identified as an aggressive PCa subtype. However, there is a lack of definite studies to elucidate the underlying mechanism of the loss of SPINK1 expression in most PCa cells except 22Rv1 cells, which are derived from a human prostatic carcinoma xenograft, CWR22R. The aim of this study was to investigate the mechanisms of SPINK1 protein positive/negative expression and its biological roles in PCa cell lines. SPINK1 mRNA was highly expressed in 22Rv1 cells compared with LNCaP, C4-2B, DU145, and PC-3 cells, and the protein was only detected in 22Rv1 cells. Among these cell lines, the wild-typeSPINK1coding sequence was only found in 22Rv1 cells, and two mutation sites, the c.194G>A missense mutation and the c.210T>C synonymous mutation, were found in other cell lines. Our further research showed that the mutations were associated with a reduction in SPINK1 mRNA and protein levels. Functional experiments indicated that SPINK1 promoted PC-3 cell proliferation, migration, and invasion, while knockdown of SPINK1 attenuated 22Rv1 cell proliferation, migration, and invasion. The wild-typeSPINK1gene can promote the malignant behaviors of cells more than the mutated ones. Cell cycle analysis by flow cytometry showed that SPINK1 decreased the percentage of cells in the G0/G1phase and increased the percentage of S phase cells. We demonstrated that the c.194G>A and c.210T>C mutations in theSPINK1gene decreased the mRNA and protein levels. The wild-typeSPINK1gene is related to aggressive biological behaviors of PCa cells and may be a potential therapeutic target for PCa.