抑癌基因SASH1在乳腺癌中结合Vimentin并负性调节Vimentin的表达

doi:10.11958/20222057

天津医药 ›› 2023, Vol. 51 ›› Issue (9): 904-908.doi: 10.11958/20222057

抑癌基因SASH1在乳腺癌中结合Vimentin并负性调节Vimentin的表达

杨娉娉¹^,²(), 张景², 陈红宇¹^,², 张淼³, 周定安², 方文¹^,^△()

1.贵州医科大学医学检验学院，贵州医科大学附属医院临床检验中心（邮编550004）
2.贵州医科大学附属医院临床研究中心
3.贵州医科大学附属医院内分泌代谢科

收稿日期:2023-01-13 修回日期:2023-04-25 出版日期:2023-09-15 发布日期:2023-09-13
通讯作者: △E-mail：fangwen@gmc.edu.cn
作者简介:杨娉娉（1997），女，硕士在读，主要从事乳腺癌的基础方面研究。E-mail：1553723457@qq.com
基金资助:
国家自然科学基金地区项目(31860319);国家自然科学基金地区项目(32060165);贵州省科技计划项目(黔科合基础-ZK[2022]一般419)

Tumor suppressor gene SASH1 binds Vimentin and negatively regulates the expression of Vimentin in breast cancer

YANG Pingping¹^,²(), ZHANG Jing², CHEN Hongyu¹^,², ZHANG Miao³, ZHOU Dingan², FANG Wen¹^,^△()

1. School of Clinical Laboratory Science, Guizhou Medical University, Clinical Laboratory Center, the Affiliated Hospital of Guizhou Medical University, Guiyang 550004, China
2. Clinical Medical Research Center, the Affiliated Hospital of Guizhou Medical University
3. Department of Endocrinology, the Affiliated Hospital of Guizhou Medical University

Received:2023-01-13 Revised:2023-04-25 Published:2023-09-15 Online:2023-09-13
Contact: △E-mail：fangwen@gmc.edu.cn

杨娉娉, 张景, 陈红宇, 张淼, 周定安, 方文. 抑癌基因SASH1在乳腺癌中结合Vimentin并负性调节Vimentin的表达[J]. 天津医药, 2023, 51(9): 904-908.

YANG Pingping, ZHANG Jing, CHEN Hongyu, ZHANG Miao, ZHOU Dingan, FANG Wen. Tumor suppressor gene SASH1 binds Vimentin and negatively regulates the expression of Vimentin in breast cancer[J]. Tianjin Medical Journal, 2023, 51(9): 904-908.

摘要/Abstract

摘要：

目的寻找参与抑癌基因SASH1调节乳腺癌发生发展的伙伴分子，并分析两者的相关性以及在乳腺癌发生发展中的作用。方法利用液相色谱-串联质谱联用（LC-MS/MS）技术和免疫共沉淀技术分析与SASH1结合并可能调节乳腺癌发生发展的分子。免疫组织化学和蛋白质免疫印迹法（Western blot）检测30例正常乳腺组织和123例乳腺癌组织SASH1和波形蛋白（Vimentin）的表达水平，并在乳腺癌中分析两者之间的相关性。Western blot检测乳腺癌细胞中敲低SASH1对Vimentin表达的影响。结果 SASH1与Vimentin存在相互作用的可能性为100%。免疫共沉淀证实SASH1与Vimentin可发生蛋白相互作用。免疫组织化学染色结果显示，与正常乳腺组织相比，乳腺癌组织中SASH1表达降低而Vimentin表达增高，且两者呈负相关（P<0.01）。Western blot实验证实在16例乳腺癌组织和对应的癌旁组织中，乳腺癌组织表现出SASH1低表达，而Vimentin高表达。在乳腺癌细胞中敲低SASH1后Vimentin的表达增高。结论 SASH1可能通过负性调节Vimentin的表达参与乳腺癌的发生发展。

关键词: 乳腺肿瘤, 波形蛋白, SASH1, 蛋白相互作用

Abstract:

Objective To investigate the partner molecules that regulate the tumor suppressor gene SASH1 in the occurrence and development of breast cancer, and to analyze the correlation and role of them in breast cancer development. Methods Liquid chromatography tandem mass spectrometry (LC-MS/MS) technology, bioinformatics and immunoprecipitation were used to analyze and confirm molecules that may bind with SASH1 and potentially regulate the occurrence and development of breast cancer. Immunohistochemistry and Western blot assay were used to detect the expression levels of SASH1 and Vimentin in normal breast tissue and breast cancer tissue in humans, and to analyze the correlation between the two in breast cancer. The effect of knocking down SASH1 on the expression of Vimentin was also examined in breast cancer cells using Western blot assay. Results LC-MS/MS technology combined with bioinformatics showed that SASH1 was more likely to bind to Vimentin. Immunoprecipitation indicated that SASH1 could bind to Vimentin. In the immunohistochemical staining of 30 cases of normal breast tissue and 123 cases of breast cancer tissue, it was found that compared with normal breast tissue, SASH1 expression was decreased and Vimentin expression was increased in breast cancer tissue. Furthermore, a negative correlation was observed between the immunohistochemical scores of the two proteins (P<0.01). Western blot assay indicated that in 16 cases of breast cancer tissue and matched adjacent tissue, the breast cancer tissue showed low expression of SASH1 and high expression of Vimentin. After knocking down SASH1 expression in breast cancer cells, the expression of Vimentin increased. Conclusion The tumor suppressor protein SASH1 may participate in the occurrence and development of breast cancer by negatively regulating the expression of Vimentin.

Key words: breast neoplasms, Vimentin, SASH1, protein-protein interaction

中图分类号:

R737.9

图/表 8

图1 免疫共沉淀验证Vimentin与SASH1蛋白相互作用 A：GFP抗体检测到GFP-SASH1和HA-Vimentin存在相互作用；B：HA抗体检测到HA-Vimentin和GFP-SASH1存在相互作用。

Fig.1 The interaction between SASH1 and Vimentin protein verified by immunoprecipitation

表1 SASH1和Vimentin在正常乳腺组织与乳腺癌组织中的表达（$\bar{x}\pm s$）

Tab.1 Expression of SASH1 and Vimentin in breast cancer tissue and normal breast tissue

组别	n	SASH1	Vimentin
正常组	30	8.93±1.57	4.80±1.04
乳腺癌组	123	6.33±1.13	7.53±1.01
t		8.579^＊＊	13.110^＊＊

图2 SASH1在正常乳腺组织与乳腺癌组织中的表达（IHC，×100）

Fig.2 Expression of SASH1 in normal breast tissue and breast cancer tissue (IHC, ×100)

图3 Vimentin在正常乳腺组织与乳腺癌组织中的表达（IHC，×100）

Fig.3 Expression of Vimentin in normal breast tissue and breast cancer tissue (IHC, ×100)

表2 SASH1和Vimentin在不同分子亚型乳腺癌组织及正常乳腺组织中的表达（$\bar{x}\pm s$）

Tab.2 Expression of SASH1 and Vimentin in different molecular subtypes of breast cancer tissue and normal breast tissue

组别	n	SASH1	Vimentin
正常组	30	8.93±1.57	4.80±1.04
Luminal A型	47	6.56±1.06^a	8.05±0.78^a
Luminal B型	34	6.73±1.08^a	7.40±1.01^ab
HER2阳性	22	6.01±1.06^a	6.54±0.94^abc
三阴性	20	5.42±0.88^abc	7.60±0.75^ad
F		35.026^＊＊	64.799^＊＊

图4 SASH1和Vimentin在乳腺癌组织和癌旁组织中的表达情况

Fig.4 Expression of SASH1 and Vimentin in breast cancer tissue and adjacent tissue

表3 癌旁组织与乳腺癌组织中SASH1和Vimentin表达水平的比较（n=16，$\bar{x}\pm s$）

Tab.3 Comparison of expression levels of SASH1 and Vimentin between adjacent tissue and breast cancer tissue

组别	SASH1/GAPDH	Vimentin/GAPDH
癌旁组织	1.47±0.40	0.08±0.08
乳腺癌组织	0.20±0.14	0.72±0.64
t	12.207^＊＊	3.984^＊＊

图5 敲低SASH1对乳腺癌细胞中Vimentin表达的影响 a与对照组比较，b与病毒空载组比较，P<0.05。

Fig.5 Effect of SASH1 knockdown on the expression of Vimentin in breast cancer cells

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抑癌基因SASH1在乳腺癌中结合Vimentin并负性调节Vimentin的表达

Tumor suppressor gene SASH1 binds Vimentin and negatively regulates the expression of Vimentin in breast cancer

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