下调表达SND1通过调控衰老相关分泌表型促进人二倍体成纤维细胞衰老的研究

孙曼; 杨晓文; 曹锋; 柯娜

doi:10.12182/20200560504

下调表达SND1通过调控衰老相关分泌表型促进人二倍体成纤维细胞衰老的研究

Downregulation of SND1 Expression Accelerates Cell Senescence of Human Diploid Fibroblasts 2BS via Modulating the SASP

摘要

摘要:
目的研究下调表达葡萄球菌核酸酶和tudor结构域1(staphylococcal nuclease and tudor domain containing 1, SND1) 对人二倍体成纤维细胞衰老的影响，并探讨其相关机制。
方法 Western blot和免疫组化分别检测SND1在年轻及衰老2BS细胞（博来霉素诱导2BS细胞老化）和老年组织（人结肠腺瘤组织）中的表达情况；免疫荧光检测SND1在年轻2BS细胞中的定位；CCK8和EDU分析检测2BS的增殖能力；集落形成分析评价2BS集落形成能力；表达芯片和RT-qPCR分析衰老相关分泌表型（SASP）表达改变；β半乳糖苷酶染色用于显示衰老的2BS细胞。
结果 SND1在衰老2BS细胞中的表达较年轻2BS细胞显著下调，且在人结肠腺瘤组织中的表达较非病变结肠组织明显下调。在年轻的2BS中，敲低SND1抑制2BS增殖和克隆形成，并出现增强的衰老相关β半乳糖苷酶染色(P<0.05)。表达芯片检测结果和RT-qPCR分析表明敲低SND1上调了SASP成分(P<0.05)。
结论本研究结果表明下调的SND1通过上调SASP表达来调控人二倍体细胞衰老。

Abstract:
Objective To investigate the effect of down-regulation of SND1 expression on senescence of human diploid fibroblasts.
Methods Western blot and immunohistochemistry were used to detect the expression of SND1 in young or senescent 2BS cells and aged tissues. Immunofluorescence was conducted to detect the localization of SND1 in young 2BS cells. CCK8 and EDU were performed to detect the proliferation of 2BS. Colony formation analysis was used to evaluate the capacity of colony formation of 2BS. Expression chip and RT-qPCR analysis were performed to detect the change of SASP expression level. β-galactosidase staining was employed to indicate the senescent 2BS cells.
Results The expression of SND1 in the senescent 2BS cells was significantly down-regulated compared with in the younger 2BS cells, and in human colon adenomas, its expression was also significantly down-regulated compared with in non-lesion colon tissues. In young 2BS, knockdown of SND1 inhibited the proliferation and colony formation of 2BS, and led to stronger senescence-associated beta-galactosidase staining (SA-β-gal). Expression chip and RT-qPCR analysis indicated that knockdown of SND1 up-regulated the expression of senescence-associated secretory phenotype components (SASP).
Conclusions Our data indicated that down-regulation of SND1 regulated human diploid cell senescence by up-regulating the expression of SASP components.

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