Construction of SIRT1 gene shRNA lentivirus vector and its effect on the proliferation of breast cancer cells

Yanbo Yue,Xin Li,Lili He,Mi Tian

doi:10.14715/cmb/2020.66.3.33

Abstract

Current experiment aimed to investigate the construction of the SIRT1 gene shRNA lentivirus vector and its effect on proliferation of breast cancer cells. Altogether 80 cases of breast cancer tissues and 80 cases of normal adjacent tissues were collected. qPCR was used for detecting SIRT1 expression. Western blot was used to detect the expression of EMT marker protein. The effect of lentivirus infected sh-SIRT1 on the cell biological function of SK-BR-3 and MDA-MB-231 cells was detected. MTT assay was used to detect cell activity, Transwell cell was used to detect cell invasion and migration, and cell apoptosis detected by flow cytometry. Compared with normal tissues adjacent to cancer, the expression of SIRT1 in cancer tissues increased significantly. Compared with human breast epithelial cells (MCF 10A), SIRT1 expression in breast cancer cells (MDA-MB-231, SK-BR-3) increased significantly. The above results showed that SIRT1 was significant greatly expressed in breast cancer. Compared with the sh-Control group, the cell activity, invasion and migration of the sh-SIRT1 group were enhanced, while cell apoptosis was weakened. In the sh-SIRT1 group infected by lentivirus, cell activity, cell invasion and migration decreased, while cell apoptosis increased. Compared with sh-Control, the expression of α-catenin, PTEN and E-cadherin in the sh-SIRT1 group in SK-BR-3 and MDA-MB-231 cells was down-regulated, while the expression of N- cadherin, β-catenin and Vimentin was up-regulated. Compared with sh-Control, the expression of α-catenin, PTEN and E-cadherin in the sh-SIRT1 group infected by lentivirus was up-regulated, while the expression of N- cadherin, β-catenin and Vimentin was down-regulated. To sum up, SIRT1 is highly expressed in breast cancer cells. The proliferation of breast cancer cells was inhibited after lentivirus infection with sh-SIRT1.

Highlights

Breast cancer has been known as the most common female malignant tumor, and even life-threatening when severe, has a major impact on women's physical and mental health [1,2,3]
The results showed that over-expression of Silent information regulator 1 (SIRT1) could promote the proliferation, migration and invasion of SK-BR-3 and MDAMB-231 cells, and inhibit cell apoptosis
Some studies have shown that SIRT1 can inhibit cell growth, proliferation and transformation by regulating glutamine metabolism, playing its role in promoting cancer

Summary

Introduction

Breast cancer has been known as the most common female malignant tumor, and even life-threatening when severe, has a major impact on women's physical and mental health [1,2,3]. Researchers have paid more and more attention to the role of molecular targeted therapy in breast cancer. Gene target therapy has progressively become a hot spot in breast cancer studies [7, 8]. Researchers have paid attention to the role of SIRT1 in the tumor. Et al found that cisplatin combined with SiRNA-SIRT 1 can reduce the expression level of drug-resistant genes in cervical cancer cells, enhancing the sensitivity of cervical cancer cells to cisplatin. Most studies believed that SIRT1 is narrowly related to the happening and development of various tumors, and it can promote the occurrence and development of breast cancer, but its exact role in breast cancer has not been clearly defined [14, 15]

Methods

Results

Conclusion