MicroRNA-383-5p acts as a potential prognostic biomarker and an inhibitor of tumor cell proliferation,migration,and invasion in breast cancer

Abstract

BACKGROUND:

MicroRNAs (miRNAs) have been reported to serve as potential biomarkers in various cancer and play important roles in tumor progression.

OBJECTIVE:

The aim of this study was to investigate the prognostic significance and functional role of miR-383-5p in breast cancer.

METHODS:

The expression levels of miR-383-5p in breast cancer tissues and cell lines were measured using quantitative real-time PCR analysis. Kaplan-Meier curve and Cox regression analysis were used to explore the prognostic significance of miR-383-5p in breast cancer. The CCK-8 assay was used to assess cell proliferation ability. Transwell assays were used to assess cell migration and invasion abilities of breast cancer cells.

RESULTS:

The expression of miR-383-5p was significantly downregulated in breast cancer tissues and cell lines, compared with that in normal tissues and normal epithelial MCF-10A cells, respectively. The expression of miR-383-5p was associated with differentiation, lymph node metastasis, and TNM stage. Patients with low miR-383-5p expression had shorter overall survival than those with high miR-383-5p expression. Overexpression of miR-383-5p significantly inhibited cell proliferation, migration, and invasion, while downregulation of miR-383-5p promoted cell proliferation, migration, and invasion in vitro. LDHA was a direct target of miR-383-5p.

CONCLUSIONS:

Taken together, miR-383-5p was downregulated in breast cancer tissues and cell lines, and overexpression of miR-383-5p inhibited cell proliferation, migration, and invasion in breast cancer cells by targeting LDHA. Based on our findings, miR-383-5p may be a prognostic biomarker and therapeutic target for breast cancer.

Keywords

miR-383-5p breast cancer prognosis proliferation migration invasion

1. Introduction

Breast cancer, one of the most frequently diagnosed malignancy with high prevalence in women worldwide, is the second and fifth leading cause of cancer death in the USA and China, respectively [1, 2]. Because breast cancer is a highly heterogeneous and complex malignancy, despite improvement in diverse therapeutic strategies, the overall survival rate of patients with advanced stages of breast cancer is still not ideal [3, 4]. Importantly, patients at advanced stage usually with metastatic lesions to the lymph nodes or other organs, such as bone marrow, lung, and liver leads to death. Therefore, it is essential to identify novel factors for the prediction and effective treatment of breast cancer and design more novel treatment strategies.

MicroRNAs (miRNAs) are a group of endogenous small noncoding RNAs (approximately 22 nucleotides in length), which regulate gene expression by binding the 3’-UTRs of the target genes, resulting in degradation or translation inhibition of target mRNAs [5, 6, 7]. Numerous studies have demonstrated that the function of miRNAs is widely involved in diverse biological processes, such as cell growth, proliferation, differentiation, migration, and invasion [8, 9]. Biological studies showed that miRNAs play an oncogenic or tumor suppressor role in the initiation and progression of human cancers [10, 11]. Recent studies also showed numerous miRNAs are aberrantly expressed in breast cancer, such as miR-329 [12], miR-27a [13], and miR-128-3p [14], indicating miRNAs play important roles in breast cancer. miR-383-5p, located in chromosome 8p22, acts as a tumor suppressor in many cancers, such as gastric cancer, colorectal cancer, and lung adenocarcinoma [15, 16, 17]. A previous study reported some breast cancer-associated miRNAs, including miR-383-5p [18]. Thus, it would be of interest to investigate the clinical significance and functional roles of miR-383-5p involved in breast cancer carcinogenesis.

Table 1
Association between miR-383-5p expression and different clinical characteristics of patients with breast cancer

Clinical characteristics	Cases	miR-383-5p expression		$P$ values
	$n=$ 124	Low ( $n=$ 64)	High ( $n=$ 60)
Age				0.202
$\leqslant$ 60	59	34	25
$>$ 60	65	30	35
Tumor size (cm)				0.075
$\leqslant$ 2	58	25	33
$>$ 2	66	39	27
ER status				0.079
Negative	47	29	18
Positive	77	35	42
PR status				0.158
Negative	45	27	18
Positive	79	37	42
HER-2 status				0.228
Negative	47	21	26
Positive	77	43	34
Differentiation				0.043
Well-moderate	82	37	45
Poor	42	27	15
Lymph node metastasis				0.025
Negative	72	31	41
Positive	52	33	19
TNM stage				0.018
I–II	65	27	38
III–IV	59	37	22

In the present study, we investigated the expression levels of miR-383-5p in breast cancer tissues and cell lines, as well as the association between the expression profile of miR-383-5p and clinicopathological characteristics of patients with breast cancer. The prognostic significance of miR-383-5p and the effects of miR-383-5p on cellular behavior were also evaluated in breast cancer patients.

2. Materials and methods

2.1 Patients and tissue samples

Paired human breast cancer tissue samples and adjacent normal tissue samples were obtained from 124 patients who received surgical resection between January 2011 and June 2013 at Shanxian Central Hospital. All surgical specimens were snap-frozen in liquid nitrogen and stored at $-$ 80 ${}^{\circ}$ C until use. No patients received chemotherapy or radiotherapy prior to the surgery section. The clinical characteristics of all patients were collected and summarized in Table 1. All patients signed written informed consent prior to participation in the present study and this study was approved by the Ethics Committee of Shanxian Central Hospital (No. SXCH-2011-005). 5-year follow-up information of all patients enrolled in this study was also obtained for further survival analysis.

2.2 RNAseq analysis

RNAseq was performed to validate the expression levels of miR-383-5p in breast cancer based on the TCGA database. TCGA RNAseq data contains 1104 primary tumors and 105 solid tissue normal samples.

2.3 Cell culture and transfection

Breast cancer cell lines (MDA-MB-231, MDA-MB-453, MCF-7, ZR-75-1) and human normal epithelial cells MCF-10A were all purchased from the Cell Bank of the Chinese Academy of Sciences (Shanghai, China). All breast cancer cells were cultured in Dulbecco’s modified Eagle’s medium (DMEM; Thermo Fisher Scientific, Waltham, USA) supplemented with 10% fetal bovine serum (FBS; Thermo Fisher Scientific, Waltham, USA) and 1% penicillin-streptomycin mixture (Gibco; Thermo Fisher Scientific, Waltham, USA) at 37 ${}^{\circ}$ C in a humidified incubator with an atmosphere of 5% CO ${}_{2}$ . For MCF-10A cells, the DMEM medium supplemented with 10% FBS, 10 $\mu$ g/ml insulin, 0.5 $\mu$ g/ml hydrocortisone, 20 ng/ml EGF, and 1% penicillin-streptomycin mixture (Gibco; Thermo Fisher Scientific, Waltham, USA).

The miR-383-5p mimic (5’-AGAUCAGAAGGUG AUUGUGGCU-3’), mimic negative control (mimic NC; 5’-ACUACUGAGUGACAGUAGA-3’), miR-383-5p inhibitor (5’-AGCCACAAUCACCUUCUGA UCU-3’), and inhibitor NC (5’-UUCUCCGAACGU GUCACGUTT-3’) were purchased from GenePharma Co., Ltd (Shanghai, China). 1 $\times$ 10 ${}^{5}$ Cells were transfected with miR-383-5p mimic, mimic NC, miR-383-5p inhibitor, or inhibitor NC; at a final concentration of 50 nM using Lipofectamine 3000 (Invitrogen; Thermo Fisher Scientific, Waltham, USA) for 48 h following to the manufacturer’s protocol. Untreated cells were used as control.

2.4 RNA extraction and quantitative real-time PCR

Total RNA was extracted from all breast cancer tissues or cells using TRIzol reagent (Invitrogen; Thermo Fisher Scientific, Waltham, USA) following the manufacturer’s protocol. After quantifying the RNA by using NanoDrop 2000 (Thermo Fisher Scientific, Waltham, USA), extracted RNA was used to synthesize complementary DNA (cDNA) using a PrimeScript RT reagent kit (Takara, Otsu, Japan). Reverse transcription primer for miR-383-5p was 5’-CTC AACTGGTGTCGTGGAGTCGGCAATTCAGTTGAGAGCCACAA-3’. The following temperature was used: 37 ${}^{\circ}$ C for 15 min, 85 ${}^{\circ}$ C for 5 sec. The expression levels of miR-383-5p were determined by quantitative real-time PCR (qRT-PCR) on an Applied Biosystems 7900 Real-time PCR System with an SYBR-Green I Master mix kit (Invitrogen; Thermo Fisher Scientific, Waltham, USA). U6 was used as an internal control. The thermocycling condition was as follows: 94 ${}^{\circ}$ C for 2 min, followed by 40 cycles of 94 ${}^{\circ}$ C for 5 s, 60 ${}^{\circ}$ C for 15 s, and 72 ${}^{\circ}$ C for 10 s. The primer sequences were: miR-383-5p forward, 5’-CACGAAAGATCAGAAGGTGA-3’, reverse, 5’-CTCAACTGGTGTCGTGGA-3’; and U6 forward, 5’- TGCGGGTGCTCGCTTCGCAGC-3’, reverse, 5’-CC AGTGCAGGGTCCGAGGT-3’. The expression of miR-383-5p in breast cancer tissue relative to the normal tissue adjacent to the tumor was calculated with the 2 ${}^{-\Delta\Delta Ct}$ method [19].

2.5 Cell proliferation assay

Cell proliferation abilities were measured using the Cell counting kit-8 (CCK-8; Dojindo, Kumamoto, Japan) assay. The breast cancer cells were transfected with miR-383-5p mimic, miR-383-5p inhibitor, or NCs as aforementioned. After 48 h of transfection, the transfected cells were seeded in 96-well plates at a density of 3 $\times$ 10 ${}^{3}$ cells/well and cultured at 37 ${}^{\circ}$ C with 5% CO ${}_{2}$ . Then 10 $\mu$ l CCK-8 reagent was added to each well at 0, 24, 48, and 72 h time points and incubated for 2 h at 37 ${}^{\circ}$ C according to the manufacturer’s protocol. The absorbance was obtained at 450 nm using a microplate reader.

2.6 Cell migration and invasion assays

Cell migratory and invasive abilities were assessed using a 24-well Transwell chamber (Corning, NY, USA) with 8 $\mu$ m pore size membranes. Different from cell migration assay, membranes were pre-coated with Matrigel (Corning, NY, USA) for invasion assay. 100 $\mu$ l transfected cells (5 $\times$ 10 ${}^{4}$ cells/well) re-suspended with the serum-free medium were placed into upper chambers. Then 500 $\mu$ l medium containing 10% FBS (Thermo Fisher Scientific, Waltham, USA) was added to the lower chamber. After 24 h of incubation, the non-migratory/non-invasive cells were removed and the migratory/invasive cells were fixed with 4% paraformaldehyde and stained with 0.1% crystal violet. Stained cells were subsequently counted in five randomly selected fields.

Figure 1.

The expression of miR-383-5p was decreased in breast cancer tissues and cell lines. (A) Expression levels of miR-383-5p were decreased in breast cancer tissues compared with adjacent normal tissues. (B) Validation expression of miR-383-5p in breast cancer using TCGA RNAseq data. (C) Expression levels of miR-383-5p were downregulated in breast cancer cell lines (MDA-MB-231, MDA-MB-453, MCF-7, and ZR-75-1) compared with normal epithelial cells MCF-10A. *** $P<$ 0.001.

2.7 Bioinformatics analysis and dual-luciferase reporter assay

Putative miR-383-5p targets were predicted using online algorithms TargetScan (http://www.targetscan. org/). MCF-7 cells were seeded in 24-well plates and incubated for 24 h prior to transfection. For the luciferase reporter assay, the wild-type (WT) 3’-UTR of LDHA or mutant (Mut) 3’-UTR of LDHA were synthesized and inserted into the pGL3 promoter vector (Promega Corporation). Then the WT or Mut 3’-UTR vectors and miR-383-5p mimics, mimic NC, miR-383-5p inhibitor, or inhibitor NC were co-transfected into MCF-7 cells using Lipofectamine 3000 (Invitrogen; Thermo Fisher Scientific, Waltham, USA) according to manufacturer’s instructions. After 48 h, a Dual-Luciferase Reporter assay system was used to measure the relative luciferase activity according to the manufacturer’s protocols (Promega Corporation). The Luciferase activity was normalized to Renilla luciferase activity.

2.8 Statistical analysis

All assays were performed at least three times. Data were analyzed using SPSS 20.0 software (SPSS, Inc., Chicago, IL, USA) and GraphPad Prism 5.0 software (GraphPad, Inc., La Jolla, CA, USA) and presented as mean $\pm$ SD. The $\chi^{2}$ test was used to analyze the association between miR-383-5p expression and clinical characteristics of patients. Student’s t-tests or one-way analysis of variance (ANOVA) was used to conduct differential comparisons of two groups or multiple groups, respectively. Kaplan-Meier method and Cox regression analyses were used to analyze the prognostic significance of miR-383-5p. A value of $P$ less than 0.05 was considered statistically significant.

3. Results

3.1 Expression of miR-383-5p in breast cancer tissues and cell lines

First, qRT-PCR was used to identify the expression levels of miR-383-5p in 124 pairs of breast cancer tissues and normal tissues. The results showed that miR-383-5p expression in breast cancer tissues was significantly lower than that in paired normal tissues ( $P<$ 0.001, Fig. 1A). Then, we validated the expression of miR-383-5p in TCGA RNAseq data and the results confirmed that miR-383-5p was downregulated in breast cancer compared with normal control ( $P<$ 0.001, Fig. 1B).

Then we detected the expression levels of miR-383-5p in breast cancer cells. We observed that the relative expression of miR-383-5p was downregulated in breast cancer cells compared with human normal epithelial MCF-10A cells ( $P<$ 0.001, Fig. 1C). Among the four breast cancer cells, the lowest expression levels of miR-383-5p were observed in MDA-MB-453 cells, followed by MDA-MB-231 and MCF-7 cells. Because of these cell lines have a relatively lower expression of miR-383-5p, they were used for subsequent experiments.

3.2 Association between miR-383-5p expression and clinical characteristics of breast cancer patients

Next, we investigated the relationships between miR-383-5p expression and clinical factors of breast cancer patients. According to the mean level of miR-383-5p expression, breast cancer patients were divided into two groups, with 64 in the low miR-383-5p expression group and 60 in the high miR-383-5p expression group. As shown in Table 1, miR-383-5p expression was significantly associated with differentiation ( $P=$ 0.043), lymph node metastasis ( $P=$ 0.025), and TNM stage ( $P=$ 0.018). No significant associations were found between miR-383-5p expression level and patients’ age, tumor size, ER status, PR status, HER-2 status ( $P>$ 0.05).

3.3 The miR-383-5p expression is correlated with survival outcomes of breast cancer patients

Considering miR-383-5p expression was associated with lymph node metastasis and TNM stage of breast cancer patients, we wonder whether miR-383-5p was correlated with the survival of patients. In order to explore the prognostic significance of miR-383-5p in breast cancer, Kaplan-Meier analysis, and log-rank tests were used to analyze the association between miR-383-5p expression and 5-year overall survival information. As expected, Kaplan-Meier survival analysis indicated that low miR-383-5p expression predicted poorer overall survival ( $P=$ 0.018, Fig. 2). The multivariate Cox regression analyses were also performed to identify factors related to the prognosis of patients. As shown in Table 2, the expression level of miR-383-5p was an independent prognostic factor (HR $=$ 2.085, 95%CI: 1.042–4.171, $P=$ 0.038).

Table 2
Multivariate Cox analysis of factors for survival of breast cancer patients

Variables	Multivariate Cox analysis
	HR	95%CI	$P$ -value
miR-383-5p	2.085	1.042–4.171	0.038
Age	0.571	0.303–1.075	0.082
Tumor size	0.686	0.365–1.290	0.242
ER status	1.013	0.513–2.004	0.970
PR status	1.220	0.629–2.367	0.556
HER-2 status	1.708	0.897–3.251	0.103
Differentiation	0.756	0.396–1.443	0.396
Lymph node metastasis	1.835	0.966–3.485	0.064
TNM stage	1.736	0.929–3.243	0.084

Figure 2.

Kaplan-Meier curve of survival time in patients with breast cancer. Patients with low miR-383-5p expression levels exhibited a shorter overall survival rate (log-rank test $P=$ 0.018).

3.4 miR-383-5p inhibits breast cancer cell proliferation, migration, and invasion

To further investigate the role of miR-383-5p in breast cancer tumorigenesis, miR-383-5p mimic, mimic NC, miR-383-5p inhibitor, or inhibitor NC was transfected into MDA-MB-231 or MDA-MB-453 cells, respectively, to conduct gain-of-function and loss-of-function assays. The qRT-PCR results showed that miR-383-5p mimic significantly upregulated miR-383-5p expression, while miR-383-5p inhibitor downregulated miR-383-5p expression, compared with control ( $P<$ 0.001, Fig. 3A). A CCK-8 assay was used to determine the effect of miR-383-5p on the proliferation of breast cancer cells in the study. As shown in Fig. 3B, overexpression of miR-383-5p significantly inhibited cell proliferation, while downregulation of miR-383-5p promoted cell growth ( $P<$ 0.05). Next, the function of miR-383-5p in breast cancer cell migration and invasion was explored using Transwell assays. As shown in Fig. 4, ectopic expression of miR-383-5p inhibited breast cancer cell migration and invasion, while downregulation of miR-383-5p promoted cell migration and invasion, compared with control ( $P<$ 0.01).

Figure 3.

Effects of miR-383-5p expression levels on proliferation in MDA-MB-231, MDA-MB-453, and MCF-7 cells. (A) Validation of miR-383-5p expression levels using qRT-PCR analysis after transient transfection. The expression of miR-383-5p-5p was significantly higher following transfection with miR-383-5p mimic and lower following transfection with miR-383-5p inhibitor compared with control (B) Proliferation of breast cancer cells was inhibited following transfection with miR-383-5p mimic and promoted following transfection with miR-383-5p inhibitor. * $P<$ 0.05, *** $P<$ 0.001.

Figure 4.

Effects of miR-383-5p on cell migration and invasion abilities in MDA-MB-231, MDA-MB-453, and MCF-7 cells. (A) Representative images of Transwell migration assay (magnification, $\times$ 200). (B) Cell migration abilities were inhibited following transfection with miR-383-5p mimic while enhanced following transfection with miR-383-5p inhibitor. (C) Representative images of Transwell invasion assay (magnification, $\times$ 200). (D) Cell invasion abilities were suppressed following transfection with miR-383-5p mimic while promoted following transfection with miR-383-5p inhibitor.** $P<$ 0.01, *** $P<$ 0.001.

3.5 LDHA was a direct target of miR-383-5p

The potential targets of miR-383-5p were investigated by bioinformatics analysis using the website TargetScan. Among these predicted targets, LDHA was selected as a potential target gene of miR-383-5p. The putative binding sites between miR-383-5p and LDHA 3’-UTR were presented in Fig. 5A. Then, luciferase reporter assay was performed to validate this interaction. The reporter plasmids were co-transfected with miR-383-5p mimic, mimic NC, miR-383-5p inhibitor, or inhibitor NC in MCF-7 cells. The luciferase reporter assay results indicated that miR-383-5p mimic transfection significantly decreased the luciferase activity of the WT 3’-UTR of LDHA, while without effects on the Mut plasmids in MCF-7 cells ( $P<$ 0.05, Fig. 5B). These findings suggested that LDHA may be a direct target of miR-383-5p.

Figure 5.

LDHA was targeted by miR-383-5p in breast cancer. (A) Putative binding sites of miR-383-5p and LDHA were predicted by TragetScan. (B) The luciferase activity was investigated in MCF-7 cells co-transfected with WT or Mut luciferase vectors and miR-383-5p mimic, mimic NC, miR-383-5p inhibitor, or inhibitor NC. Overexpression of miR-383-5p decreased the luciferase activity of the WT 3’-UTR of LDHA, while without effects on the Mut plasmids in MCF-7 cells * $P<$ 0.05, ** $P<$ 0.01.

4. Discussion

Accumulating evidence has demonstrated that miRNAs involved in tumor initiation and progression of cancers, which are still under investigation and act as a new class of promising cancer biomarkers [20, 21]. Numerous miRNAs were also reported to be associated with breast cancer [22, 23]. For instance, miR-876-5p was downregulated in breast cancer cells and suppressed breast cancer cell proliferation, migration, and invasion by targeting TFAP2A [22]. Serum miR-21 and miR-125b were upregulated in breast cancer patients and both of them were identified as the novel, noninvasive predictive markers for neoadjuvant chemotherapy response and prognosis in breast cancer [23]. However, there is still lacking molecular markers available for clinical application in breast cancer.

Many studies have shown miR-383-5p was abnormally regulated in various cancers [15, 16, 17]. As most studies have indicated, miR-383 acts as a tumor suppressor in inhibiting cell proliferation, migration, and enhancing chemo-sensitivity in multiple cancers [24]. However, the potential role of miR-383-5p in breast cancer is still unclear. In the present study, miR-383-5p was downregulated in breast cancer tissues and cells, compared with adjacent normal tissues and cells, respectively. Then, we validated the expression of miR-383-5p in TCGA RNAseq data and the results confirmed that miR-383-5p was downregulated in breast cancer compared with normal control. Furthermore, the majority of patients with low miR-383-5p expression exhibited poor differentiation, positive lymph node metastasis, and advanced TNM stage. The above results suggest that miR-383-5p may be involved in the development of breast cancer. Furthermore, given the dysregulated miR-383-5p expression in breast cancer tissues, we evaluated the prognostic significance of miR-383-5p in breast cancer. Kaplan-Meier results showed that patients with low miR-383-5p expression had shorter overall survival rates. The results of the present study suggested that miR-383-5p was an independent prognostic biomarker for predicting overall survival of patients with breast cancer. The above results of the present study were consistent with those of previous studies demonstrating that miR-383-5p was downregulated in cancer tissues and acted as a potential marker of the therapeutic target [17, 25]. For instance, miR-383 is downregulated in lung adenocarcinoma tissues and acts as a potential prognostic biomarker and therapeutic target for lung adenocarcinoma [17]. In glioma, downregulation of miR-383 promotes cell invasion by targeting IGF1R and miR-383 may act as a useful diagnostic marker or therapeutic target [25].

Numerous studies have demonstrated that aberrant expression of miRNAs is associated with diverse biological processes, such as proliferation, migration, and invasion [26, 27, 28]. In breast cancer, aberrant miRNAs expression has been reported involved in numerous physiological processes and these miRNAs may be potential therapeutic targets for the treatment of breast cancer, such as miR-206 [29], miR-211-5p [30], and miR-590-3p [31]. According to the expression levels of miR-383-5p in breast cancer tissues and cells, we speculate that miR-383-5p may have a functional role as a tumor suppressor in breast cancer. To further explore whether miR-383-5p functions as a suppressor in breast cancer progression, the effects of miR-383-5p on breast cancer cell proliferation, migration, and invasion were assessed. Ectopic expression of miR-383-5p significantly inhibited cell proliferation, migration, and invasion in transfected MDA-MB-231, MDA-MB-453, and MCF-7 cell lines, while inhibition of miR-383-5p promoted these cellular behaviors. Therefore, these results suggested that miR-383-5p may play an inhibitory role in breast cancer. In colon cancer, miR-383 was downregulated in cancer tissues and inhibited cell proliferation, migration, and invasion of colon cancer cells through the regulation of target gene APRIL [32]. In hepatocellular carcinoma (HCC), miR-383 was downregulated in HCC tissues and cell lines and might inhibit HCC cell proliferation, invasion and glycolysis by targeting LDHA [33]. LDHA has been reported upregulated in multiple types of cancers and played crucial role in cancer cell proliferation, invasion, and metastasis of various tumors [34, 35]. In breast cancer, LDHA was reported upregulated and knockdown of LDHA could inhibit the migration and invasion of breast cancer cells [36, 37, 38]. In the present study, bioinformatics analysis prediction showed there are putative binding sites between miR-383-5p and LDHA 3’-UTR. And dual-luciferase reporter assay results showed that LDHA was the direct target of miR-383-5p. Based on previous studies and the present study results, we speculate that miR-383-5p may inhibit breast cancer cell proliferation, migration, and invasion by targeting LDHA. However, the detailed molecular mechanisms underlying the role of miR-383-5p in breast cancer still need to be investigated in future researches.

Taken together, the results of the present study indicated that miR-383-5p is downregulated in breast cancer tissues and cell lines and downregulation of miR-383-5p is associated with poor differentiation, positive lymph node metastasis, advanced TNM stages, and poor survival in patients with breast cancer. The results of the present study revealed that overexpression of miR-383-5p may suppress breast cancer proliferation, migration, and invasion by targeting LDHA, suggesting that miR-383-5p/LDHA axis may be a novel therapeutic target and strategy for the treatment of patients with breast cancer.

Footnotes

Conflict of interest

The authors declare no conflict of interest.

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MicroRNA-383-5p acts as a potential prognostic biomarker and an inhibitor of tumor cell proliferation,migration,and invasion in breast cancer

Abstract

BACKGROUND:

OBJECTIVE:

METHODS:

RESULTS:

CONCLUSIONS:

Keywords

1. Introduction

Table 1 Association between miR-383-5p expression and different clinical characteristics of patients with breast cancer

2.1 Patients and tissue samples

2.2 RNAseq analysis

2.3 Cell culture and transfection

2.4 RNA extraction and quantitative real-time PCR

2.5 Cell proliferation assay

2.6 Cell migration and invasion assays

2.8 Statistical analysis

3. Results

3.1 Expression of miR-383-5p in breast cancer tissues and cell lines

3.2 Association between miR-383-5p expression and clinical characteristics of breast cancer patients

3.3 The miR-383-5p expression is correlated with survival outcomes of breast cancer patients

Table 2 Multivariate Cox analysis of factors for survival of breast cancer patients

Footnotes

Conflict of interest

References

Table 1
Association between miR-383-5p expression and different clinical characteristics of patients with breast cancer

Table 2
Multivariate Cox analysis of factors for survival of breast cancer patients