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

doi:10.11958/20222057

Abstract

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

CLC Number:

R737.9

Figures/Tables 8

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

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^＊＊

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

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

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^＊＊

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

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^＊＊

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

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