Long non-coding RNA MIDEAS-AS1 inhibits growth and metastasis of triple-negative breast cancer via transcriptionally activating NCALD

Luo, Dan; Liang, Yiran; Wang, Yajie; Ye, Fangzhou; **, Yuhan; Li, Yaming; Han, Dianwen; Wang, Zekun; Chen, Bing; Zhao, Wen**g; Wang, Lijuan; Chen, **; Jiang, Liyu; Yang, Qifeng

doi:10.1186/s13058-023-01709-1

Long non-coding RNA MIDEAS-AS1 inhibits growth and metastasis of triple-negative breast cancer via transcriptionally activating NCALD

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Published: 28 September 2023

Volume 25, article number 109, (2023)
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Long non-coding RNA MIDEAS-AS1 inhibits growth and metastasis of triple-negative breast cancer via transcriptionally activating NCALD

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Dan Luo¹^na1,
Yiran Liang¹^na1,
Yajie Wang¹^na1,
Fangzhou Ye¹,
Yuhan **¹,
Yaming Li¹,
Dianwen Han¹,
Zekun Wang¹,
Bing Chen³,
Wen**g Zhao³,
Lijuan Wang³,
** Chen¹,
Liyu Jiang¹ &
…
Qifeng Yang^1,2,3

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Abstract

Background

Triple-negative breast cancer (TNBC) is a subtype of breast cancer with higher aggressiveness and poorer outcomes. Recently, long non-coding RNAs (lncRNAs) have become the crucial gene regulators in the progression of human cancers. However, the function and underlying mechanisms of lncRNAs in TNBC remains unclear.

Methods

Based on public databases and bioinformatics analyses, the low expression of lncRNA MIDEAS-AS1 in breast cancer tissues was detected and further validated in a cohort of TNBC tissues. The effects of MIDEAS-AS1 on proliferation, migration, invasion were determined by in vitro and in vivo experiments. RNA pull-down assay and RNA immunoprecipitation (RIP) assay were carried out to reveal the interaction between MIDEAS-AS1 and MATR3. Luciferase reporter assay, Chromatin immunoprecipitation (ChIP) and qRT-PCR were used to evaluate the regulatory effect of MIDEAS-AS1/MATR3 complex on NCALD.

Results

LncRNA MIDEAS-AS1 was significantly downregulated in TNBC, which was correlated with poor overall survival (OS) and progression-free survival (PFS) in TNBC patients. MIDEAS-AS1 overexpression remarkably inhibited tumor growth and metastasis in vitro and in vivo. Mechanistically, MIDEAS-AS1 mainly located in the nucleus and interacted with the nuclear protein MATR3. Meanwhile, NCALD was selected as the downstream target, which was transcriptionally regulated by MIDEAS-AS1/MATR3 complex and further inactivated NF-κB signaling pathway. Furthermore, rescue experiment showed that the suppression of cell malignant phenotype caused by MIDEAS-AS1 overexpression could be reversed by inhibition of NCALD.

Conclusions

Collectively, our results demonstrate that MIDEAS-AS1 serves as a tumor-suppressor in TNBC through modulating MATR3/NCALD axis, and MIDEAS-AS1 may function as a prognostic biomarker for TNBC.

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Background

Breast cancer is one of the most common malignant tumors that has serious effects on the health of women worldwide [3,4,5]. TNBC is characterized by higher rates of relapse, greater metastatic potential, and shorter overall survival compared with other major breast cancer subtypes [4, 5]. Therapeutic methods for TNBC patients usually include surgery, chemotherapy, radiotherapy, and immunotherapy [6,7,8,9]. Clinically, although significant progress in the treatment of TNBC over the last decade, recurrence and metastasis remain the principal causes of mortality in patients with this disease [Full size image

Table 1 Correlation between MEDIA-AS1 mRNA expression and clinicopathological characteristics in 91 triple-negative breast cancer patients

Full size table

MIDEAS-AS1 reduced the proliferation, migration and invasion of TNBC cells in vitro

To explore the potential roles of MIDEAS-AS1, we transfected the MIDEAS-AS1 overexpression plasmids into MDA-MB-231 and MDA-MB-468 cells, and the overexpression efficiency was determined by qRT-PCR (Additional file 1: Fig. S2A, S2B). Results from MTT assay indicated that the proliferation ability of MDA-MB-231 and MDA-MB-468 cells with MIDEAS-AS1 overexpression was reduced compared to control group (Fig. 2A). Meanwhile, overexpression of MIDEAS-AS1 also inhibited the proliferation of breast cancer organoids (Fig. 2B). Moreover, overexpression of MIDEAS-AS1 repressed cell colony-forming activity (Fig. 2C-D). Additionally, flow cytometry was used to detect the apoptosis rate and cell cycle after different treatments. Our results indicated that MIDEAS-AS1 overexpression led to remarkably increased apoptosis rate (Fig. 2E) and cell number in G1 phase (Additional file 1: Fig. S2C). On the other hand, MIDEAS-AS1 knockdown promoted the proliferation and colony-formation abilities of TNBC cells (Fig. 2A, D). Furthermore, the apoptosis rate was obviously decreased and the number of cells in G1 phase was less in MIDEAS-AS1 knockdown group compared to that in control group (Fig. 2E, Additional file 1: Fig. S2C). We then investigated the effect of MIDEAS-AS1 on TNBC cell migration and invasion using transwell assays and wound healing assay in MDA-MB-231 and MDA-MB-468 cells. The results of transwell assays showed a significant reduction in the migratory and invasive abilities of MDA-MB-231 and MDA-MB-468 cells after MIDEAS-AS1 overexpression (Fig. 2F). Furthermore, wound healing assay indicated that the wound closure area of MDA-MB-231 and MDA-MB-468 cells was dramatic decreased following MIDEAS-AS1 overexpression (Additional file 1: Fig. S2D). Consistently, MIDEAS-AS1 knockdown led to increased migratory and invasive abilities of TNBC cells (Fig. 2G). Given that epithelial-mesenchymal transition (EMT) is one of the major mechanisms for cancer metastasis, we further examined the effect of MIDEAS-AS1 on the expression of EMT-related marker proteins by Western blot. The results showed that N-cadherin, vimentin, fibronectin and MMP9 proteins were downregulated, while the E-cadherin protein was upregulated in MDA-MB-231 and MDA-MB-468 cells after overexpression of MIDEAS-AS1 (Fig. 2H). On the contrary, N-cadherin, vimentin, fibronectin and MMP9 proteins were upregulated, and the E-cadherin protein was downregulated in TNBC cells after MIDEAS-AS1 knockdown (Fig. 2I). Therefore, MIDEAS-AS1 could regulate the EMT process to modulate TNBC metastasis. Collectively, these results indicated that MIDEAS-AS1 played critical roles in suppressing cell proliferation and mobility of TNBC cells.

MIDEAS-AS1 directly interacts with MATR3 to carry out its function

It had been suggested that the regulatory mechanisms of lncRNAs were closed associated with their subcellular localization [25]. Therefore, we firstly detected the subcellular localization of MIDEAS-AS1 in TNBC cells. Following isolation of the nuclear and cytoplasmic RNA in MDA-MB-231 and MDA-MB-468 cells, qRT-PCR analysis demonstrated that MIDEAS-AS1 was mainly located in the nucleus (Fig. 3A). Furthermore, FISH assay also obtained the similar results (Fig. 3B), indicating the potential role of MIDEAS-AS1 in transcriptional regulation through acting as a scaffold for TFs [25, 27]. To identify proteins interacted with MIDEAS-AS1, RNA pull-down assay was performed. We incubated MDA-MB-231 cell lysates with biotinylated MIDEAS-AS1 or its antisense RNA transcribed in vitro, and the silver staining and mass spectrometry (MS) identified MATR3 as one of the major proteins in the MIDEAS-AS1 pull-down precipitations (Fig. 3C). Moreover, mass spectrometry analysis found that MIDEAS-AS1 interacted with MATR3 at “DLSAAGIGLLAAATQSLSMPASLGR” and “YQLLQLVEPFGVISNHLILNK” peptide sequences (Fig. 3D). The specific binding between MATR3 protein and MIDEAS-AS1 was further confirmed by RNA pull down following Western blot (Fig. 3E). Then, we performed RNA immunoprotein (RIP) assay using flag antibodies and revealed that MIDEAS-AS1 was significantly enriched in MATR3 immunoprecipitations (Fig. 3F). Meanwhile, RNA FISH technology combined with immunofluorescence analysis demonstrated the co-localization of MIDEAS-AS1 and MATR3 protein in MDA-MB-231 cells (Fig. 3G). To identify the specific binding regions between MIDEAS-AS1 and MATR3, we constructed vectors containing full-length MIDEAS-AS1 or three truncated sequences (Fig. 3H). The RNA pull down assay indicated that deleting the sequence of 217–439 bp (Δ2 vector) could abolish the MIDEAS-AS1-MATR3 interaction, which was consist with the prediction results obtained from catRAPID database (Additional file 1: Fig. S3A). Furthermore, we wonder whether MIDEAS-AS1 could regulate the expression of MATR3. Significantly, overexpression or knockdown of MIDEAS-AS1 shows no effect on the RNA and protein expression levels of MATR3 (Additional file 1: Fig. S3B), indicating that MIDEAS-AS1 was not involved in the post-transcriptional regulation of MATR3. Collectively, these results revealed that MIDEAS-AS1 specifically interacted with MATR3 in TNBC cells.

MIDEAS-AS1 affects the progression and metastasis of TNBC by promoting NCALD expression

Given the critical role of nuclear matrix-associated protein MATR3 in transcriptional regulation, we wonder whether MIDEAS-AS1 could regulate the expression of downstream genes through interacting with MATR3 to further affect the progression and metastasis of TNBC. We performed an RNA-sequencing analysis on MIDEAS-AS1-overexpressing and control cells, and 471 differentially expressed genes were identified (Fig. 4A-B), including 252 up-regulated and 219 down-regulated genes. Meanwhile, the top 20 up-regulated genes are shown in Fig. 4C. Due to the tumor-suppressive role of MIDEAS-AS1 in TNBC, we integrated the down-regulated genes in breast cancer tissues from TCGA database and the up-regulated genes after MIDEAS-AS1 overexpression in TNBC cells, and then, 9 candidate genes were selected as potential downstream targets of MIDEAS-AS1 (Fig. 4D). It has been reported in the literature that antisense lncRNA may affect the expression of sense gene [28], so we also examined the influence of MIDEAS-AS1 on its sense gene (MIDEAS) by qRT-PCR. However, the results showed that MIDEAS-AS1 did not affect the expression of MIDEAS (Additional file 1: Fig. S4B). Meanwhile, the qRT-PCR results revealed that the expression of AF131215.5, CXCL1, and NCALD was remarkably increased after MIDEAS-AS1 overexpression and reduced after MIDEAS-AS1 knockdown (Additional file 1: Fig. S4A). CXCL1 (C-X-C motif chemokine ligand 1) was the most abundant chemokine secreted by TAMs, and CXCL1 could promote migration, invasion ability, and EMT in breast cancer [29, 30]. AF131215.5 was a lncRNA whose function remained unclear in cancer [31, 32]. Interestingly, NCALD (neurocalcin delta) expression was lower in lung adenocarcinoma tissues [33], and patients with higher NCALD levels exhibited a higher survival rate [34, 35]. Among these genes, we hypothesized that NCALD, the tumor suppressor gene, might be a potential target of MIDEAS-AS1 in breast cancer. Significantly, NCALD expression was markedly downregulated in breast cancer tissues compared with normal tissues (Additional file 1: Fig. S4C-D). Moreover, immunohistochemistry (IHC) showed higher expression of NCALD in normal tissues than breast cancer tissues according to Human Protein Atlas database (Additional file 1: Fig. S4E). These results indicated that NCALD might play a significant tumor suppressor role in breast cancer. Therefore, NCALD was selected as the functional downstream mediator for MIDEAS-AS1-mediated cell migration and metastasis in TNBC. Significantly, the qRT-PCR and Western blot results revealed that the expression of NCALD was remarkably increased after MIDEAS-AS1 or MATR3 overexpression and reduced after MIDEAS-AS1 or MATR3 knockdown in MDA-MB-231 and MDA-MB-468 cells (Fig. 4E-F, Additional file 1: Fig. S4E), indicating the regulatory effect of MIDEAS-AS1 and MATR3 on the expression of NCALD. Furthermore, rescue experiments were performed to demonstrate the causal link between MIDEAS-AS1, MATR3 and NCALD. The qRT-PCR analysis indicated that MIDEAS-AS1 overexpression promoted the expression of NCALD, while MATR3 knockdown could weaken the increased tendency of NCALD expression caused by overexpression of MIDEAS-AS1 in MDA-MB-231 and MDA-MB-468 cells (Fig. 4G). Furthermore, we found that the increasing trend of NCALD protein level caused by MIDEAS-AS1 overexpression could be attenuated by MATR3 knockdown in MDA-MB-231 and MDA-MB-468 cells (Fig. 4H). Then, we further investigated the transcriptional regulation effect of MIDEAS-AS1 and MATR3 on the expression of NCALD. The dual luciferase reporter assay showed that overexpression of MIDEAS-AS1 or MATR3 substantially increased the luciferase activity of NCALD vectors, while knockdown of MIDEAS-AS1 or MATR3 significantly inhibited the luciferase activity of NCALD vectors in HK293T cells (Fig. 4I). Furthermore, rescue experiments revealed that the promoter activity of NCALD was activated by MIDEAS-AS1 overexpression, which was inhibited after co-transfection with si-MATR3 (Fig. 4J). These results indicated that MIDEAS-AS1 could regulate the expression of NCALD through modulating the function of MATR3. To identify the specific binding regions of MIDEAS-AS1 and MATR3 on the NCALD promoter, we ectopically expressed full-length NCALD promoter (−2000 to + 100 bp) as well as five mutants: the truncated NCALD-1 (−2000 to −1564 bp) mutant, the truncated NCALD-2 (−1563 to −1094 bp) mutant, the truncated NCALD-3 (−1093 to −654 bp) mutant, the truncated NCALD-4 (−653 to −269 bp) mutant, and the truncated NCALD-5 (−268 to + 100 bp) mutant (Fig. 4K), and a luciferase assay was performed. We found that the luciferase activity was most significantly increased in −1563 bp to −1094 bp, indicating the presence of positive regulatory elements, which enhanced NCALD transcription in this region (Fig. 4L). In addition, the ChIP followed by qPCR assays was subsequently performed in Flag-MATR3 transfected breast cancer cells with or without MIDEAS-AS1 overexpression to determine the effect of MIDEAS-AS1 on MATR3 recruitment to the NCALD promoter. Consistent with the luciferase assay, MIDEAS-AS1-MATR3 complex was identified to significantly bound to −1563 bp to −1094 bp sites of the NCALD promoter region in MDA-MB-231 and MDA-MB-468 cells (Fig. 4M). These findings suggested that MIDEAS-AS1-MATR3 complex could enhance NCALD transcription by directly binding to its promoter.

NCALD inhibits TNBC cell proliferation, migration, and invasion in vitro

Previous studies had found that NCALD was not only involved in cell apoptosis, cell cycle progression and other biological processes in several cancers [36], but also associated with the prognosis of cancers [34, 35]. However, there is no report about the function of NCALD in breast cancer. Our above results revealed downregulated expression of NCALD in breast cancer tissues and breast cancer cells, indicating the tumor-suppressive role of NCALD in breast cancer. To confirm the function of NCALD, we transfected NCALD overexpressing vectors and si-NCALD into MDA-MB-231 and MDA-MB-468 cells, and the overexpression and knockdown efficiency were determined by qRT-PCR and Western blot assays (Fig. 5A-B). The MTT and colony formation assay results indicated that NCALD overexpression reduced TNBC cell proliferation and colony formation abilities (Fig. 5C, D). Additionally, the apoptosis rate was remarkably increased and the number of cells in G1 phase was increased after NCALD overexpression (Fig. 5F, Additional file 1: Fig. S5A). On the other hand, NCALD knockdown led to significantly increased proliferation, colony formation abilities of TNBC cells (Fig. 5C, E). Moreover, the apoptosis rate was decreased and the number of the cells in G1 phase was less in NCALD knockdown cells than that in control cells (Fig. 5F, Additional file 1: Fig. S5A). Additionally, transwell assay and wound healing assay revealed that NCALD overexpression significantly reduced TNBC cell migration and invasion abilities (Fig. 5G, Additional file 1: Fig. S5B), while NCALD knockdown led to increased migration and invasion abilities in TNBC cells (Fig. 5H, Additional file 1: Fig. S5B). These data demonstrated that NCALD served as a suppressive functional factor in TNBC progression.

Previous literature reported that NCALD might be related to ERK1/2 signaling pathway, NF-κB signaling pathway, TGF-β signaling pathway and immune response pathway in ovarian cancer [35]. We further investigated the potential molecular mechanism caused by the change of NCALD in TNBC cells. Western blot analysis indicated that overexpression of NCALD would significantly inhibited phosphorylate p-65, but not TGF-β and, p-ERK1/2 in MDA-MB-231 and MDA-MB-468 cells (Fig. 5I), while NCALD knockdown brought about opposite results (Fig. 5J). Collectively, our results demonstrated that NCALD inhibited TNBC cell proliferation, migration, and invasion by suppressing NF-κB signaling pathways.

MIDEAS-AS1 regulates TNBC progression through regulating the expression of NCALD

Our previous results have showed that the association between MIDEAS-AS1 and MATR3 played significant role in initiating NCALD transcription. To further confirm whether MIDEAS-AS1 exerts tumor-suppressive functions via modulating NCALD, we co-transfected si-NCALD and MIDEAS-AS1-overexpressed plasmid in TNBC cell lines. The MTT and colony formation experiments indicated that NCALD knockdown remarkably rescued the proliferation ability of MDA-MB-231 and MDA-MB-468 cells inhibited by the overexpression of MIDEAS-AS1 (Fig. 6A-B). Moreover, the transwell and wound healing assays showed that NCALD knockdown could partially recover the cell migration and invasion abilities reduced by MIDEAS-AS1-overexpression (Fig. 6C-D). Moreover, Western blot analysis revealed that NF-κB pathway-related proteins were decreased after overexpression of MIDEAS-AS1, and that was increased after co-transfection of MIDEAS-AS1 overexpression plasmid and si-MATR3 (Fig. 6E). Taken together, MIDEAS-AS1 associates with MATR3 to initiate NCALD transcription and inhibits NF-κB signaling pathway, which further affects the TNBC progression.

MIDEAS-AS1 overexpression inhibits TNBC progression and metastasis in vivo

In order to further evaluate the biological function of MIDEAS-AS1 in vivo, a subcutaneous xenograft model was first constructed. The MDA-MB-231 cells stably transfected with MIDEAS-AS1 overexpressing vectors or control vectors were subcutaneously injected to the flanks of nude mice. The result showed that the tumor weight and tumor volume were significantly inhibited in xenografts of MIDEAS-AS1 overexpressing group compared with those in the control group (Fig. 7A–C). Furthermore, immunohistochemistry (IHC) assays confirmed that MIDEAS-AS1 overexpression caused decreased Ki67 expression and increased NCALD expression, but did not affect MATR3 expression (Fig. 7D-E). In addition, we further constructed pulmonary metastasis model through intravenously injecting MDA-MB-231 cells stably expressing MIDEAS-AS1 or control vectors to compare the metastatic abilities in vivo. The results showed that mice injected with MIDEAS-AS1-overexpressing TNBC cells had no or fewer metastatic foci compared with the control group (Fig. 7F–H). Together, these results indicated that MIDEAS-AS1 played a critical role in inhibiting TNBC progression and metastasis.

Discussion

Breast cancer is one of the most common malignancies among women worldwide and is the major cause of most cancer-related deaths. There are several explanations for the high mortality rate of breast cancer, with metastasis of vital organs thought to be the main cause [37]. As reported in the literature, TNBC is the most challenging subtype of breast cancer with higher rates of relapse and greater metastatic potential compared to other breast cancer subtypes [4, 38, 39]. Despite the treatment methods for patients with metastatic breast cancer are complicated, the curative effects are unsatisfactory in the clinical practice. Therefore, it is of great significance to investigate the molecular mechanism of TNBC metastasis and identify novel prognostic predictors for accurate diagnosis and prediction of prognosis.

It is well known that lncRNA is abnormally expressed in many different types of cancer and is involved in the regulation of tumor development and progression [40,41,42]. Recently, various studies have focused on the functions and regulations of lncRNAs to search novel diagnostic and therapeutic targets for cancer treatment. In this study, we explored the potential role and molecular mechanism of MIDEAS-AS1 in TNBC progression. We determined that MIDEAS-AS1 was significantly downregulated in TNBC tissues compared to the non-TNBC tissues, and low expression of MIDEAS-AS1 was associated with poor prognosis in TNBC. Functional studies revealed that MIDEAS-AS1 suppressed proliferation, migration and invasion, metastasis and promoted apoptosis of TNBC cells in vitro, indicating a tumor suppressor role in TNBC. Moreover, MIDEAS-AS1 inhibited TNBC tumor progression and lung metastasis in vivo by xenograft model. However, the regulatory mechanism of MIDEAS-AS1 involved in TNBC progression was still unclear and worthy of further exploration.

It is reported that the localization of lncRNAs within the cell is the primary determinant of their molecular functions [43]. Increasing evidence suggests that cytoplasmic lncRNAs can regulate gene expression by modulating mRNA stability, translation process or participating in mRNA post-transcriptional regulation as ceRNAs [19, 44]. Meanwhile, the nuclear lncRNAs also participate in several biological processes, including chromatin organization, and transcriptional and post-transcriptional gene expression, and acting as structural scaffolds of nuclear domains [24]. Here, we found that MIDEAS-AS1 was mainly located in the nucleus based on cell cytoplasmic/nuclear fractionation and RNA FISH assays. Previous studies have reported that lncRNAs could play important role in transcription by recruiting corresponding proteins [45, 46]. Therefore, MIDEAS-AS1 might exert its function by recruiting transcription complexes and further enhance or inhibit gene transcription. Then, RNA pull-down followed by mass spectrometry showed the binding potential between MATR3 and MIDEAS-AS1, which was further confirmed by RIP assay. Moreover, we also found that MIDEAS-AS1 and MATR3 were co-localization in TNBC cells through RNA FISH technology combined with immunofluorescence. Subsequently, we identified the specific binding region between MIDEAS-AS1 and MATR3. However, we found that the expression level of MIDEAS-AS1 did not affect the RNA and protein levels of MATR3, making us speculating that MIDEAS-AS1 might regulated the function of MATR3 in TNBC cells.

It is reported that MATR3 is an abundant nuclear protein that binds with DNA and RNA [47, 48], allowing MATR3 to play crucial roles in RNA splicing and gene transcription [49,50,51]. To further explore the downstream genes regulated by the MIDEAS-AS1-MATR3 complex, we analyzed the RNA-seq results and 471 DEGs were revealed. Following integration with TCGA database, we finally identified 9 candidate genes as potential direct downstream targets of MIDEAS-AS1 and MATR3. NCALD, a member of the neuronal calcium sensors protein family [52], caught our attention due to the remarkable changes after MIDEAS-AS1 overexpression or knockdown. Studies have found that NCALD was associated with the prognosis of several cancers, including non-small cell lung cancer, ovarian cancer, colorectal cancer, indicating its clinical potential as a prognostic biomarker [35, 36]. However, the function role and molecular mechanism of NCALD in breast cancer has not been reported. In this study, we found that NCALD was downregulated in breast cancer tissues. Moreover, the expression of NCALD could be regulated by MATR3 in TNBC cells. Significantly, rescue experiment revealed that MATR3 knockdown attenuated the increasing trend of NCALD expression caused by MIDEAS-AS1 overexpression in TNBC cells. Subsequently, dual luciferase reporter assays and ChIP-qPCR indicated that MIDEAS-AS1 and MATR3 complex could bind to NCALD promoter to regulate NCALD transcription.

We further investigated whether NCALD modulate the functional effect of MIDEAS-AS1 in TNBC. Functional experiments revealed that NCALD overexpression inhibited cell proliferation, migration, and invasion in TNBC cells, suggesting the suppressive functional role of NCALD in TNBC progression. Previous study has reported that the TGF-β, NF-κB and ERK signaling pathways were associated with NCALD expression in epithelial ovarian cancer [35]. Interestingly, Western blot analysis showed that overexpression of NCALD inhibited the expression of the NF-κB pathway-associated proteins, indicating that NCALD inhibited progression of TNBC possibly by suppressed NF-κB signaling pathway. Furthermore, the rescue experiments showed that NCALD knockdown could partially rescue the proliferation and migration ability of TNBC cells inhibited by overexpression of MIDEAS-AS1. Western blot analysis also revealed that MIDEAS-AS1 and MATR3 could not only affect the NCALD expression, but also significantly modulate the NF-κB signaling pathway. Therefore, our study revealed that MIDEAS-AS1 regulated TNBC progression by recruiting the MATR3 to initiate NCALD transcription and suppressing NF-κB signaling pathways (Fig. 7I).

Conclusions

In this study, we identified MIDEAS-AS1 as a novel tumor suppressor in TNBC, and higher expression of MIDEAS-AS1 was associated with better prognosis of TNBC patients. Mechanistically, we revealed that MIDEAS-AS1 inhibited TNBC progression through activating NCALD transcription via recruiting MATR3. Our study illustrates a novel mechanism involved in TNBC progression and suggests that MIDEAS-AS1 might serve as a prognostic biomarker for TNBC patients.

Availability of data and materials

The data that support the findings of this study are available on request from the corresponding author upon reasonable request.

Abbreviations

TNBC:: Triple-negative breast cancer
lncRNAs:: Long non-coding RNAs
RIP:: RNA immunoprecipitation
ChIP:: Chromatin immunoprecipitation
OS:: Overall survival
PFS:: Progression-free survival
ceRNA:: Competing endogenous RNAs
GEO:: Gene Expression Omnibus
TCGA:: The Cancer Genome Atlas

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Funding

This work was supported by National Key Research and Development Program (No. 2020YFA0712400), Special Foundation for Taishan Scholars (No. ts20190971), Special Support Plan for National High Level Talents (Ten Thousand Talents Program W01020103), Foundation from Clinical Research Center of Shandong University (No. 2020SDUCRCA015), Qilu Hospital Clinical New Technology Develo** Foundation (No. 2019-3), National Natural Science Foundation of China (No. 81902697).

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Dan Luo, Yiran Liang and Yajie Wang have contributed equally to this work.

Authors and Affiliations

Department of Breast Surgery, General Surgery, Qilu Hospital, Cheeloo College of Medicine, Shandong University, Wenhua ** Road No. 107, **an, 250012, Shandong, China
Dan Luo, Yiran Liang, Yajie Wang, Fangzhou Ye, Yuhan **, Yaming Li, Dianwen Han, Zekun Wang, ** Chen, Liyu Jiang & Qifeng Yang
Pathology Tissue Bank, Qilu Hospital of Shandong University, **an, 250012, Shandong, China
Qifeng Yang
Research Institute of Breast Cancer, Shandong University, **an, 250012, Shandong, China
Bing Chen, Wen**g Zhao, Lijuan Wang & Qifeng Yang

Authors

Dan Luo
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Yiran Liang
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Yajie Wang
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Fangzhou Ye
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Yuhan **
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Yaming Li
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Dianwen Han
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Zekun Wang
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Bing Chen
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Wen**g Zhao
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Lijuan Wang
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** Chen
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Liyu Jiang
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Qifeng Yang
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Contributions

DL, YL, LJ, QY conceived the study. FY, YJ, YL, XC helped to perform the experiments. DL, DH, ZW analyzed the data. BC, WZ, LW collected clinical samples. DL, YL, YW wrote the paper. DL, YL, LJ, QY revised the paper. All authors have read and approved the final manuscript.

Corresponding authors

Correspondence to Liyu Jiang or Qifeng Yang.

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This study was achieved approval by the Ethical Committee on Scientific Research of Shandong University Qilu Hospital (KYLL-2016(KS)-140).

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Not applicable.

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The authors declare no competing interests.

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Luo, D., Liang, Y., Wang, Y. et al. Long non-coding RNA MIDEAS-AS1 inhibits growth and metastasis of triple-negative breast cancer via transcriptionally activating NCALD. Breast Cancer Res 25, 109 (2023). https://doi.org/10.1186/s13058-023-01709-1

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Received: 30 May 2023
Accepted: 11 September 2023
Published: 28 September 2023
DOI: https://doi.org/10.1186/s13058-023-01709-1

Long non-coding RNA MIDEAS-AS1 inhibits growth and metastasis of triple-negative breast cancer via transcriptionally activating NCALD