Zhu Lin, Sun Yangyan, Wu Qunying, Zhang Chenxia, Ling Jing
Department of Central laboratory, Affiliated Jiangyin Hospital of South-East University, Jiangyin, China.
Department of Obstetrics and Gynecology, Affiliated Jiangyin Hospital of South-East University, Jiangyin, China.
Ann Clin Lab Sci. 2020 Jan;50(1):24-30.
Calpain 6 (CAPN6) is one of the calcium-dependent intracellular nonlysosomal proteases that are dysregulated in uterine leiomyomas (UtLMs). However, its function and mechanism in UtLMs is still unknown.
The correlation between CAPN6 expression and UtLMs was analyzed by the Gene Expression Omnibus (GEO). The expression of CAPN6 and Rac1 was detected by quantitative real-time PCR (qPCR) and western blot analysis. Cell proliferation ability was analyzed by a Cell Counting Kit-8 (CCK-8) assay. Cell apoptosis was detected by flow cytometry.
CAPN6 was overexpressed in UtLMs compared with uterine smooth muscle cells (UtSMCs). The downregulation of CAPN6 resulted in decreased cell proliferation and increased apoptosis of UtLMs. Furthermore, mechanical investigations revealed that these inhibitory effects were correlated with Rac1/PAK1 signaling pathways. Silencing the expression of CAPN6 resulted in decreased Rac1 and phospho-PAK1. On the other hand, upregulated Rac1 expression could reverse the reduced phosphorylation of PAK1 induced by CAPN6 silencing.
This data suggests that CAPN6 regulates UtLMs proliferation and apoptosis while being mediated through the Rac1/PAK1 signaling pathway.
钙蛋白酶6(CAPN6)是一种钙依赖性细胞内非溶酶体蛋白酶,在子宫平滑肌瘤(UtLMs)中表达失调。然而,其在UtLMs中的功能和机制仍不清楚。
通过基因表达综合数据库(GEO)分析CAPN6表达与UtLMs之间的相关性。采用定量实时PCR(qPCR)和蛋白质印迹分析检测CAPN6和Rac1的表达。通过细胞计数试剂盒-8(CCK-8)检测分析细胞增殖能力。采用流式细胞术检测细胞凋亡。
与子宫平滑肌细胞(UtSMCs)相比,CAPN6在UtLMs中过表达。CAPN6的下调导致UtLMs细胞增殖减少和凋亡增加。此外,机制研究表明,这些抑制作用与Rac1/PAK1信号通路相关。沉默CAPN6的表达导致Rac1和磷酸化PAK1减少。另一方面,上调Rac1表达可逆转CAPN6沉默诱导的PAK1磷酸化降低。
这些数据表明,CAPN6通过Rac1/PAK1信号通路调节UtLMs的增殖和凋亡。