Research on the Negatively Regulation of Long Intergenic Non-Coding RNA 00210 to miR-424-5p in Breast Cancer Cells

2021 ◽  
Vol 11 (3) ◽  
pp. 520-526
Author(s):  
Xingguo Cui ◽  
Weiguang Xu

In this work, we investigate the expression of long intergenic non-coding RNA 00210 (LINC00210) and its effects on the behavior of breast cancer cells. To this end, we measured LINC00210 and miR-424-5p expression using RT-qPCR. Bioinformatics, dual luciferase report experiments, and RT-qPCR were applied to determine the potential function of LINC00210 in the regulation of miR-424-5p. Four groups of T-47D cells were set up: si-NC, si-LINC00210, si-LINC00210 + anti-miR-NC, and si-LINC00210 + anti-miR-424-5p. Then, cell viability, apoptosis, migration, and invasion were detected, respectively. Western blot analysis was applied to measure the expression levels of E-cadherin, N-cadherin, Bax, and Bcl-2. Our results showed that breast cancer tissue highly expressed LINC00210 and slightly expressed miR-424-5p, and that a direct binding function of LINC00210 to miR-424-5p existed. Furthermore, many of the behaviors of T-47D cells in the si-LINC00210 group were affected, including reductions in cell viability, migration and invasion abilities, as well as decreased expressions of LINC00210, Ki67, Bcl-2, and N-cadherin, an increased apoptosis rate, and increased expressions of miR-424-5p, E-cadherin, and Bax. In addition, in comparison with the si-LINC00210 + anti-miR-NC group, the cell behaviors of T-47D cells in the si-LINC00210 + anti-miR-424-5p group were affected, including increased cell viability, migration and invasion abilities, and expressions of Ki67, Bcl-2, and N-cadherin, but reductions in E-cadherin and Bax. The results demonstrated the inhibitory effects of LINC00210 on T-47D cells, as well as the negative regulation of LINC00210 on miR-424-5p, leading to cell apoptosis. The results imply the potential value of LINC00210 as a therapeutic target for breast cancer.

2022 ◽  
Vol 22 ◽  
Author(s):  
Wei Shi ◽  
Lu Qi ◽  
Xiong-Bin You ◽  
Yu-Chi Chen ◽  
Yu-Lian Xu ◽  
...  

Background: Shenling Baizhu Powder (SBP), a famous Traditional Chinese Medicine (TCM) formulation, has been widely used in the adjuvant treatment of cancers, including breast cancer. This study aims to identify potential new targets for breast cancer treatment based on the network pharmacology of SBP. Methods: By analyzing the relationship between herbs and target proteins, potential targets of multiple herbs in SBP were identified by network pharmacology analysis. Besides, by comparing the data of breast cancer tissue with normal tissue, upregulated genes in two breast cancer expression profiles were found. Thereafter, the expression level and prognosis of activator of heat shock protein 90 (HSP90) ATPase activity 1 (AHSA1) were further analyzed in breast cancer by bioinformatics analysis, and the network module of AHSA1 binding protein was constructed. Furthermore, the effect of knocking down AHSA1 on the proliferation, migration, and invasion of breast cancer cells was verified by MTT, clone formation assay, and transwell assay. Results: Vascular endothelial growth factor A (VEGFA), intercellular adhesion molecule 1 (ICAM1), chemokine (C-X-C motif) ligand 8 (CXCL8), AHSA1, and serpin family E member 1 (SERPINE1) were associated with multiple herbs in SBP. AHSA1 was remarkably upregulated in breast cancer tissues and positively correlated with poor overall survival and disease metastasis-free survival. Furthermore, knockdown of AHSA1 significantly inhibited the migration and invasion in MCF-7 and MDA-MB-231 breast cancer cells but had no obvious effect on proliferation. In addition, among the proteins that bind to AHSAl, the network composed of proteasome, chaperonin, and heat shock proteins is closely connected, and these proteins are associated with poor prognosis in a variety of cancers. Conclusion: AHSA1 is positively correlated with breast cancer progression and might act as a novel therapeutic target for breast cancer.


2021 ◽  
Vol 50 (10) ◽  
pp. 3015-3033
Author(s):  
Wee Yee Tan ◽  
Boon Yin Khoo ◽  
Ai Lan Chew

Atypical chemokine receptor proteins are termed ‘decoy proteins’ as their binding to the respective ligands does not lead to a typical signaling pathway but intercepts the action of chemokines. This method of chemokine activity regulation may also function in tumor suppression. D6 and DARC (Duffy Antigen Receptor for Chemokines) have been reported as decoy chemokine receptors in cancer studies. Purified Pichia-expressed D6 and DARC, produced in-house, were used in cell-based studies to test their biological activities. Cell viability tests showed that recombinant D6 and DARC did not affect cell viability significantly, suggesting that they were not involved in breast cancer cell death. Wound healing assays showed that the presence of recombinant D6 or DARC at 10 µg/mL optimally inhibited the migration of breast cancer cells. ELISA showed an inverse relationship between the recombinant proteins and CCL2 levels in the treated cells. Migration assay using Boyden chamber demonstrated the function of the recombinant proteins in inhibiting chemotaxis activity of treated cells. Invasion assay showed the ability of the recombinant proteins in inhibiting the invasion property of treated cells. Comparison of single and combinatorial effects of the recombinant proteins showed that the combination of D6 and DARC at a 1:1 ratio (10 µg/mL) is most effective in reducing CCL2 levels and inhibiting the migration and invasion of treated cells. It was shown that the purified Pichia-expressed recombinant D6 and DARC are the negative regulators of breast cancer cell migration and invasion, and the inhibition effects were greater when they were used in combination.


2020 ◽  
Vol 477 (19) ◽  
pp. 3803-3818 ◽  
Author(s):  
Sulagna Sanyal ◽  
Payel Mondal ◽  
Sabyasachi Sen ◽  
Sumita Sengupta (Bandyopadhyay) ◽  
Chandrima Das

hTERT, the catalytic component of the human telomerase enzyme, is regulated by post-translational modifications, like phosphorylation and ubiquitination by multiple proteins which remarkably affects the overall activity of the enzyme. Here we report that hTERT gets SUMOylated by SUMO1 and polycomb protein CBX4 acts as the SUMO E3 ligase of hTERT. hTERT SUMOylation positively regulates its telomerase activity which can be inhibited by SENP3-mediated deSUMOylation. Interestingly, we have established a new role of hTERT SUMOylation in the repression of E-cadherin gene expression and consequent triggering on the epithelial-mesenchymal-transition (EMT) program in breast cancer cells. We also observed that catalytically active CBX4, leads to retention of hTERT/ZEB1 complex onto E-cadherin promoter leading to its repression through hTERT-SUMOylation. Further through wound healing and invasion assays in breast cancer cells, we showed the tumor promoting ability of hTERT was significantly compromised upon overexpression of SUMO-defective mutant of hTERT. Thus our findings establish a new post-translational modification of hTERT which on one hand is involved in telomerase activity maintenance and on the other hand plays a crucial role in the regulation of gene expression thereby promoting migration and invasion of breast cancer cells.


Nutrients ◽  
2019 ◽  
Vol 11 (5) ◽  
pp. 1116 ◽  
Author(s):  
Ariana Acevedo-Díaz ◽  
Gabriela Ortiz-Soto ◽  
Ivette J. Suárez-Arroyo ◽  
Astrid Zayas-Santiago ◽  
Michelle M. Martínez Montemayor

Breast cancer (BC) is the second leading cause of cancer death among women worldwide. The main cause of BC morbidity and mortality is the invasiveness capacity of cancer cells that may lead to metastasis. Here, we aimed to investigate the therapeutic efficacy of Ganoderma lucidum extract (GLE)—a medicinal mushroom with anticancer properties—on BC motility via the Rac/Lamellipodin pathway. GLE treatment effects were tested on MDA-MB-231 breast cancer cells. The effects were tested on cell viability, migration and invasion. Pulldowns, immunoblotting, and immunofluorescence were used to measure Rac activity and the expression of proteins involved in cell migration and in lamellipodia formation, respectively. As a result, GLE suppressed BC cell viability, migration, and invasion capacity. GLE impaired Rac activity, as well as downregulated Lamellipodin, ENA/VASP, p-FAK (Tyr925), Cdc42, and c-Myc expression. Lamellipodia formation was significantly reduced by GLE. In conclusion, we demonstrate that GLE reduces Rac activity and downregulates signaling molecules involved in lamellipodia formation. These novel findings serve as basis for further studies to elucidate the potential of GLE as a therapeutic agent regulating the Rac/Lamellipodin pathway in BC metastasis.


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