MicroRNA-935 acts as a prognostic marker and promotes cell proliferation,migration,and invasion in colorectal cancer

Abstract

Colorectal cancer (CRC) is a common cause of cancer-related deaths worldwide. MicroRNA-935 (miR-935) has been reported to be involved in several cancers. In the present study, we aimed to investigate the role of miR-935 in the progression of CRC. The expression levels of miR-935 in CRC tissues and cells were detected using reverse transcription-quantitative polymerase chain reaction (RT-qPCR). The prognostic significance of miR-935 was identified using Kaplan-Meier survival analysis and Cox regression assay. Cell proliferation capacity was detected by Cell Counting Kit-8 (CCK-8) assay. Cell migration and invasion capacities were determined by Transwell assay in CRC cells. Bioinformatics analysis and luciferase reporter assays were used to confirm the direct target of miR-935. The expression of miR-935 was increased in CRC tissues and cells. Overexpression of miR-935 was significantly associated with lymph node metastasis and TNM stage. Overexpression of miR-935 in patients predicted shorter overall survival compared to low miR-935 expression. The expression of miR-935 was indicated to be an independent prognostic factor for CRC patients. In addition, overexpression of miR-935 in CRC cells promoted cell proliferation, migration, and invasion, whereas inhibition of miR-935 suppressed cell proliferation, migration, and invasion. Bioinformatics and luciferase assays revealed that INPP4A is a direct target of miR-935. Our findings suggest that miR-935 functions as an oncogene and promotes CRC progression. INPP4A is a potential target of miR-935. And miR-935 may represent a prognostic biomarker and potential therapeutic strategy for CRC treatment.

Keywords

Colorectal cancer microRNA-935 prognosis proliferation migration invasion

Table 1
Correlation between miR-935 expression levels and clinical features in CRC patients

Parameters	Cases no.	miR-935 expression		$P$
	( $n=$ 132)	Low ( $n=$ 63)	High ( $n=$ 69)
Age				0.210
$<$ 50	70	37	33
$\geqslant$ 50	62	26	36
Gender				0.336
Male	79	35	44
Female	53	28	25
Tumor size				0.228
$<$ 5 cm	68	29	39
$\geqslant$ 5 cm	64	34	30
Tumor location				0.721
Colon	65	30	35
Rectum	67	33	34
Differentiation				0.105
Well $+$ Moderate	72	39	33
Poor	60	24	36
Lymph node metastasis				0.016
Negative	59	35	24
Positive	73	28	45
TNM stage				0.006
I–II	57	35	22
III–IV	75	28	47

1. Introduction

Colorectal cancer (CRC) remains a common malignant tumor of the digestive tract worldwide and accounts for the fifth leading cause of cancer-related cause of mortality in China [1]. The incidence of CRC has increased rapidly in recent years in China. Because CRC often occurs sporadically and insidious, the patients often diagnosed with advanced stage and more with recurrence and metastasis [2]. Metastasis is the leading cause of death in patients with CRC, and nearly 25% of patients are found with metastasis in the process of initial diagnosis [3]. Over the past decade, substantial improvements in diagnosis and therapies have helped to cure the disease of patients at early stages, but the overall survival of patients at advanced stages is still unsatisfactory [4]. Thus, it is of great importance to identify reliable molecular markers that may be beneficial for improving the prognosis of CRC and the response to the treatments.

MicroRNAs (miRNAs or miRs) are a class of small non-coding RNAs of 18–25 nucleotides in length that have been shown suppress gene expression at the post-transcriptional level by binding to the 3’-UTR of the target gene [5]. A number of studies have demonstrated that miRNAs function as oncomiRs or tumor suppressor miRNAs in the progression of various cancers and play a crucial role in several biological processes, including proliferation, migration, invasion, apoptosis, and differentiation [6, 7, 8]. In recent years, studies have shown that several miRNAs are involved in the proliferation, migration, and invasion of CRC [9, 10]. MicroRNA-935 (miR-935) has been demonstrated playing an important role in various cancers, such as non-small lung cancer, pancreatic cancer, bladder cancer [11, 12, 13]. A previous study has also found that miR-935 is significantly increased in CRC tissues, however, its role in CRC progression and clinical therapy still unclear [14].

In the present study, we investigated the expression of miR-935 in CRC tissues and cell lines and analyzed its prognostic value in CRC. In addition, we investigated the effects of miR-135 on biological behaviors of CRC cells.

2. Materials and methods

2.1 CRC patients and tissue specimens

Between January 2010 and December 2012, a total of 132 pairs of CRC tissues and matched adjacent normal tissues were collected from 132 CRC patients (including 101 left-sided CRC and 31 right-sided CRC patients) who underwent surgical resection at Shengli Oilfield Central Hospital. Besides, 43 pairs of CRC tissues and adjacent normal tissues from another group CRC patients (including 30 left-sided CRC and 13 right-sided CRC patients) between February 2011 and October 2012 were used as validation samples. No patients had received chemotherapy or radiotherapy before surgery. All tissue specimens were snap-frozen and stored at $-$ 80 ${}^{\circ}$ C for RNA extraction. The histopathology of the disease was confirmed by at least two pathologists. Clinical characteristics of CRC patients are collected and listed in Table 1. Written informed consent from all participants was signed before the operation. The current studies were approved by the Ethical Committee of the Shengli Oilfield Central Hospital, and in line with the Declaration of Helsinki. The 5-year follow-up information was calculated by telephone.

2.2 Cell lines and transfection

In this study, four CRC cell lines (SW480, SW620, HCT116, and CaCO ${}_{2}$ ) and a normal colon epithelial cell line (FHC) were purchased from American Type Culture Collection (Manassas, VA, USA) and cultured in Dulbecco’s modified Eagle’s medium (DMEM, Gibco; Thermo Fisher Scientific, Inc., Waltham, MA, USA) or L15 medium (Gibco; Thermo Fisher Scientific, Inc., Waltham, MA, USA) supplemented with 10% FBS (Gibco; Thermo Fisher Scientific, Inc.) at 37 ${}^{\circ}$ C in a humidified environment of 5% CO ${}_{2}$ . Cells were seeded into 6-well plates at a density of 1 $\times$ 10 ${}^{5}$ per well and performed transfection after 24 h. Cells were transfected with miR-135 mimic, mimic negative control (NC), miR-135 inhibitor or inhibitor NC using Lipofectamine 2000 (Invitrogen, Life Technologies) following to the manufacturer’s instructions and continued to be incubated for 48 h. Untreated cells were used as control.

2.3 RNA extraction and real-time quantitative polymerase chain reaction (qRT-PCR)

Total RNA was extracted from CRC tissues and cultured cells using TRIzol reagent (Invitrogen, Thermo Fisher Scientific, Inc.) following the manufacturer’s protocol. RNA purity was performed by measuring absorbance at 260 nm and 280 nm. 100 ng RNA was used for complementary DNA (cDNA) synthesis using RevertAid First Strand cDNA Synthesis kit (Thermo Fisher Scientific, Inc.). qRT-PCR was performed using a TaqMan microRNA assay kit (Applied Biosystems-Thermo Fisher Scientifc Inc., Waltham, MA, USA) on an Applied Biosystems 7900 Real-Time PCR system (Applied Biosystems, Foster City, CA, USA). All data were analyzed using the 2 ${}^{-\Delta\Delta Ct}$ methods. U6 was used as the endogenous control to detect miR-935 expression.

2.4 Cell proliferation assay

The proliferation capacity of CRC cells was measured using Cell Counting Kit-8 (CCK-8; Beyotime Institute of Biotechnology, Jiangsu, China) assay according to the manufacturer’ instruction. Cells were incubated into 96-well plates at a density of 3 $\times$ 10 ${}^{3}$ cells per well, then incubated for 0, 24, 48, and 72 h in a 37 ${}^{\circ}$ C, 5% CO ${}_{2}$ incubator. The CCK-8 assay was performed at each time point to detect cell proliferation capacity. Briefly, CCK-8 was added into wells and incubation was continued for an additional 1 h. The absorbance was measured by a microplate reader at 450 nm.

2.5 Migration and invasion assays

The migratory and invasive abilities of CRC cells were assessed through migration and invasion assays by using a pore size of 8 $\mu$ m Transwell chamber (BD Biosciences, Franklin Lakes, NJ, USA). Transwell chambers pre-coated with Matrigel (BD Biosciences, Franklin Lakes, NJ, USA) were used for the invasion assay, while the migration assay was performed using Transwell chamber without Matrigel. The transfected cells (5 $\times$ 10 ${}^{4}$ cells) were inoculated in the upper chamber of the transwell with 100 $\mu$ L serum-free growth medium. As a chemoattractant, the lower chamber was filled with 500 $\mu$ L complete medium containing 10% FBS. After 24 h of incubation at 37 ${}^{\circ}$ C, migrated or invasive cells were fixed and stained. Then the number of cells was counted under a microscope.

Figure 1.

The expression of miR-935 was upregulated in CRC tissues and cell lines. A. The expression of miR-935 was higher in CRC tissues than adjacent normal tissues ( ${}^{***}P<$ 0.001). B. The expression of miR-935 was increased in CRC cell lines compared with a normal cell ( ${}^{**}P<$ 0.01, ${}^{***}P<$ 0.001).

Figure 2.

Kaplan-Meier survival curve of the overall survival of 132 CRC patients.

2.6 Bioinformatics analysis and dual-luciferase reporter assay

TargetScan (http://www.targetscan.org/), miRDB (http://www.mirdb.org), and miRanda (http://www.microrna.org/) database were carried out to predict the putative target gene INPP4A of miR-935. To confirm the prediction, a luciferase reporter assay was carried out in SW620 cells, which have a better biological function. To perform the dual luciferase reporter assay, wild-type or mutant INPP4A 3’-UTR luciferase reporter vector was designed, the 3’UTR sequence of INPP4A containing miR-935 binding sites was cloned downstream of the firefly luciferase gene in a pGL3-promoter vector (Promega, Madison, WI, USA). SW620 cells (1 $\times$ 10 ${}^{5}$ cells/well) were seeded on a 24-well plate for the luciferase reporter assay. The cells were then co-transfected with wild-type (WT) or mutant (Mut) 3’-UTR vectors together with miR-935 mimic, miR-935 inhibitor or NCs using Lipofectamine 2000 (Invitrogen, Life Technologies, USA) according to the manufacturer’s instructions. 48 h after transfection, luciferase activities were analyzed using the Dual-Luciferase Reporter Assay System (Promega, Madison, WI, USA) and was normalized to Renilla luciferase.

2.7 Statistical analysis

All statistical analyses were carried out using SPSS 20.0 software (SPSS, Inc., Chicago, IL, USA) and GraphPad Prism 5.0 software (GraphPad Software, Inc., Chicago, USA). Data are described with mean $\pm$ standard deviation (SD) and analyzed by $\chi^{2}$ tests, Student’s $t$ -test or one-way ANOVA followed by Tukey’s post-hoc test. The clinical significance of miR-935 in CRC was determined using the Kaplan-Meier analysis and Cox regression analysis. Differences were considered statistically significant when $P$ was less than 0.05. Each experiment was replicated at least three times.

3. Results

3.1 Expression of miR-935 in CRC tissues and cells

To determine the role of miR-935 in CRC progression, we first detected miR-935 expression in 132 paired CRC tissues and normal tissues. As shown in Fig. 1A, the expression of miR-935 was significantly higher in CRC tissues than that in the adjacent normal tissues ( $P<$ 0.001). Consistent with the results obtained in tissue specimens, miR-935 expression was markedly increased in all CRC cells compared with the normal colon epithelial FHC cells (all $P<$ 0.01, Fig. 1B).

3.2 The association between miR-935 expression and clinical parameters of CRC patients

We next analyzed the relationship between miR-935 expression and clinical parameters of CRC patients. All CRC patients were divided into miR-935 high and low expression groups according to the mean value of the miR-935 expression in tumor tissues. As shown in Table 1, the level of miR-935 was significantly associated with lymph node metastasis ( $P=$ 0.016) and TNM stage ( $P=$ 0.006). However, there were no statistically significant correlations between miR-935 expression and other parameters, including age, gender, tumor size, tumor location, and differentiation (all $P>$ 0.05).

3.3 The prognostic value of miR-935 in CRC

We then used Kaplan-Meier analysis to analyze the association between the miR-935 expression and the prognosis of patients with CRC according to miR-935 expression levels and patients survival data. The results of the Kaplan-Meier survival curve showed that patients with high miR-935 expression had worse overall survival than those with low miR-935 expression ( $P=$ 0.007, Fig. 2). Multivariate analyses were conducted to investigate the prognostic value of miR-935 in CRC using Cox regression analysis. As shown in Table 2, miR-935 expression level was an independent prognostic factor for 5-year overall survival in CRC patients (HR $=$ 1.911, 95% CI $=$ 1.019–3.584, $P=$ 0.044).

Table 2
Multivariate Cox analysis of clinical parameters in relation to overall survival

Characteristics	Multivariate analysis
	HR	95% CI	$P$
miR-935	1.911	1.019–3.584	0.044
Age	0.774	0.435–1.377	0.383
Gender	0.698	0.371–1.315	0.266
Tumor size (cm)	0.808	0.450–1.451	0.476
Tumor location	1.804	0.969–3.360	0.063
Differentiation	0.696	0.394–1.231	0.213
Lymph node metastasis	1.172	0.662–2.073	0.586
TNM stage	1.731	0.930–3.222	0.083

3.4 Validation of miR-935 expression and clinical significance in an independent validation group

To further identify the expression and clinical prognostic significance of miR-935 in CRC, the expression of miR-935 was detected and Kaplan-Meier analysis was established for CRC patients in the independent validation group. The expression of miR-935 was significantly higher in CRC tissues than in adjacent normal tissues ( $P<$ 0.001, Fig. 3A). The Kaplan-Meier survival analysis results revealed that high expression of miR-935 was correlated with poor overall survival ( $P=$ 0.042, Fig. 3B).

Figure 3.

Validation of miR-935 expression and clinical significance in an independent validation group. A. Expression of miR-935 in an independent validation tissue samples of CRC patients ( ${}^{***}P<$ 0.001). B. Upregulation of miR-935 expression was associated with poor overall survival of CRC patients (log-rank $P=$ 0.042).

3.5 miR-935 promotes the proliferation, migration, and invasion in CRC cells

To investigate the role of miR-935 in CRC, we transfected miR-935 mimic, mimic NC, miR-935 inhibitor or inhibitor NC into CRC cell line SW480 and SW620, which had relatively higher miR-935 expression levels. The results showed the expression of miR-935 was significantly upregulated in cells following transfection with miR-935 mimics, while its expression was significantly inhibited in cells that transfected with miR-935 inhibitor, respectively ( $P<$ 0.001, Fig. 4A). CCK-8 analysis results showed that overexpression of miR-935 increased cell proliferation, while knockdown of miR-935 significantly inhibited cell proliferation, compared with control ( $P<$ 0.05, Fig. 4B). Transwell assays were used to evaluate the migration and invasion capacities of CRC cells. The results also indicated upregulation of miR-935 by miR-935 mimic promotes the migration and invasion capacities of SW480 and SW620 cells, whereas downregulation of miR-935 by miR-935 inhibitor significantly inhibited cell migration and invasion capacities, compared with control ( $P<$ 0.05, Fig. 5A–D).

Figure 4.

Effects of miR-935 on CRC cell proliferation in SW480 and SW620 cells. A. Expression of miR-935 in cells transfected with miR-935 mimic was increased, while in cells transfected with miR-935 inhibitor was decreased, compared to controls ( ${}^{***}P<$ 0.001). B. The effect of miR-935 overexpression or inhibition on the proliferation of cells ( ${}^{*}P<$ 0.05).

Figure 5.

Effects of miR-935 on CRC cell migration and invasion in SW480 and SW620 cells. A. Representative migration images (magnification, $\times$ 200). B. The effect of miR-935 on CRC cell migration ( ${}^{*}P<$ 0.05, ${}^{***}P<$ 0.001). C. Representative invasion images (magnification, $\times$ 200). D. The effect of miR-935 on CRC cell invasion ( ${}^{*}P<$ 0.05, ${}^{***}P<$ 0.001).

3.6 INPP4A is a direct target of miR-935 in CRC cells

In this study, we used TargetScan, miRDB, and miRanda and found that 3’-UTR of INPP4A has a binding site for miR-935 (Fig. 6A). To confirm the prediction, we performed a dual luciferase reporter assay. Results showed that miR-935 overexpression decreased the luciferase activity of the reporter carrying the WT 3’-UTR sequences in SW620 cells, while inhibition of miR-935 expression promoted the luciferase activity of the reporter carrying the WT 3’-UTR sequences in SW620 cells ( $P<$ 0.001, Fig. 6B). However, miR-935 mimic and miR-935 inhibitor did not affect the luciferase activities of SW620 cells in the Mut 3’-UTR group (all $P>$ 0.05, Fig. 6B). These results suggested that miR-935 may directly target INPP4A 3’-UTR in CRC cells.

Figure 6.

INPP4A was a target of miR-935 in CRC. A. Computer prediction of miR-935 binding sites in the 3’-UTR of the human INPP4A is shown. B. The relative luciferase activity of INPP4A 3’UTR in SW620 cells cotransfected with wild-type (WT) or mutated (Mut) INPP4A with miR-935 mimics, inhibitor or respective negative controls ( ${}^{***}P<$ 0.001).

4. Discussion

Due to rapid progression and advanced tumor presentation at the time of diagnosis, CRC is associated with a high mortality rate, which is closely correlated with the tumor metastasis, especially the liver metastasis [15, 16]. Approaches to improve the prognosis of CRC patients may contribute to the establishment of personalized treatment [17]. Thus, it is important to identify novel and effective biomarkers for CRC. Recent studies have demonstrated that miRNAs are aberrantly expressed and regarded as a diagnostic or prognostic gene in cancers, which may help predict the prognosis of patients and treatment strategies [18, 19, 20]. For instance, a recent study indicated that high expression of miR-98 is associated with longer survival and is a good prognostic factor in acute myeloid leukemia patients treated with chemotherapy alone [21].

In the present study, we investigated the expression of miR-935 in the CRC tissues and cell lines. The results showed that miR-935 was upregulated in CRC tissues and cell lines. In addition, we found that overexpression of miR-935 was associated with clinical parameters, including lymph node metastasis and TNM stage. These results suggested miR-935 is involved in the development and progression of CRC. And overexpression of miR-935 may be associated with aggressive progression in CRC. What’s more, Kaplan-Meier analysis results indicated that patients with higher miR-935 expression levels had a shorter overall survival duration compared with those with lower miR-935 expression. And Cox regression analysis results showed that miR-935 is an independent prognostic marker of overall survival for CRC patients. To further confirm the results above, the expression of miR-935 levels and clinical information of 43 CRC patients were analyzed from the independent validation group. The results showed that the expression of miR-935 was upregulated in CRC tissues compared with normal tissues, and overexpression of miR-935 has a potential prognostic value in CRC.

A number of studies have demonstrated that miR-935 expression was upregulated in various cancers, such as pancreatic carcinoma, bladder cancer, liver cancer, and gastric cancer [12, 13, 22, 23]. In pancreatic carcinoma, the expression of miR-935 was markedly increased in cancer tissues and cells, and with INPP4A might serve as potential targets in the therapy of pancreatic cancer [12]. Liu et al. indicated that miR-935 is upregulated in liver cancer tissues and cells, and promotes liver cancer cell proliferation and migration by targeting SOX7 [22]. miR-935 was also found upregulated in gastric cancer tissues and cell lines, as well as promoted cell proliferation and tumorigenesis through targeting SOX7 [23]. These studies indicated miR-935 is involved in the progression of many cancers, considering the overexpression of miR-935 in CRC, we speculated that miR-935 may also be involved in the progression of CRC.

Numerous studies have also revealed that some miRNAs closely associated with tumor proliferation, migration, and invasion in various cancers, including CRC [24, 25, 26]. A study by Wang et al. demonstrated the miR-769 was downregulated in CRC tissues and cell lines and inhibited CRC cell proliferation, apoptosis, migration, and invasion by targeting CDK1 [27]. In another study, miR-19a was found to be as an oncomiR role and promoted CRC proliferation and migration in CRC by targeting TIA1 [28]. Considering the role of miRNAs in the progression of various cancers, we analyzed the effects of miR-935 on the proliferation, migration, and invasion in CRC cells to investigate the biological functions of miR-935 in the development and progression of CRC in the present study. The results suggested miR-935 functions as an oncogene and promotes cell proliferation, migration, and invasion, which may promote CRC progression. Intriguingly, we used bioinformatics analyses and predicted that INPP4A was a direct target gene of miR-935 in CRC. A study by Wang et al. showed that upregulation of miR-935 promotes pancreatic cancer cell proliferation and migration and inhibits cell apoptosis by targeting INPP4A [12]. A recent study by Wang R et al. indicated that overexpression of miR-940 may promote bladder cancer cell progression via targeting INPP4A and activating the Wnt/ $\beta$ -catenin pathway [29]. In addition, a study indicated that miR-4286 promotes esophageal carcinoma development by targeting INPP4A to evoke the JAK2/STAT3 pathway activation [30]. Based on these studies, we speculate miR-935 may also promote CRC proliferation, migration, and invasion by targeting INPP4A. Although our study has demonstrated that miR-935 is involved in the development and progression of CRC, the detailed molecular mechanisms of miR-935 in CRC is still unclear. Further investigation is required to explore the precise mechanisms of miR-935 in CRC progression. In addition, the sample size is limited, further studies with larger research cohorts are also needed.

Taken together, this study demonstrated that miR-935 was significantly upregulated in CRC, which was closely related to poor prognosis of CRC patients and was a potential prognostic biomarker. miR-935 may promote CRC proliferation, migration, and invasion by targeting INPP4A. The present study indicated miR-935 may act as an oncogene in CRC progression and has the potential to improve targeted treatment for CRC.

Footnotes

Acknowledgments

The authors thank the colleagues in the Pathology Unit for providing the data in the independent validation group.

Conflict of interest

The authors declare that they have no conflict of interests.

References

Zhang

Chen

and Li

, The current status of treatment for colorectal cancer in China: A systematic review, Medicine (Baltimore) 96 (2017), e8242.

Tauriello

D.V.

Calon

Lonardo

and Batlle

, Determinants of metastatic competency in colorectal cancer, Mol Oncol 11 (2017), 97–119.

Xin

Wang

and Liu

, miR-196a-5p promotes metastasis of colorectal cancer via targeting IkappaBalpha, BMC Cancer 19 (2019), 30.

Brenner

Kloor

and Pox

C.P.

, Colorectal cancer, Lancet 383 (2014), 1490–502.

Lee

and Vasudevan

, Post-transcriptional stimulation of gene expression by microRNAs, Adv Exp Med Biol 768 (2013), 97–126.

Yong-Ming

Ai-Jun

Xiao-Yue

Jian-Wei

Chen

and Ye

, miR-449a: a potential therapeutic agent for cancer, Anticancer Drugs 28 (2017), 1067–78.

Hou

Fang

Song

Sun

Liu

and Chen

, MicroRNA-18a promotes proliferation and metastasis in oral squamous cell carcinoma via targeting KLF4, Cancer Biomark (2018).

Wang

R.H.

L.Y.

and Zhou

S.H.

, The role of gene sculptor microRNAs in human precancerous lesions, Onco Targets Ther 11 (2018), 5667–75.

Chen

Gao

Liang

Qiao

Duan

Shi

Zhen

Zhang

Zhu

and Han

, miR-203a-3p promotes colorectal cancer proliferation and migration by targeting PDE4D, Am J Cancer Res 8 (2018), 2387–401.

10.

Wang

Rui

and Xu

M.Q.

, miR-410 acts as an oncogene in colorectal cancer cells by targeting dickkopf-related protein 1 via the Wnt/beta-catenin signaling pathway, Oncol Lett 17 (2019), 807–14.

11.

Peng

Cai

Zhao

and Chen

, Knockdown of miR935 increases paclitaxel sensitivity via regulation of SOX7 in nonsmallcell lung cancer, Mol Med Rep 18 (2018), 3397–402.

12.

Wang

Feng

Jiang

and Zuo

, Upregulation of MicroRNA-935 Promotes the Malignant Behaviors of Pancreatic Carcinoma PANC-1 Cells via Targeting Inositol Polyphosphate 4-Phosphatase Type I Gene (INPP4A), Oncol Res 25 (2017), 559–69.

13.

Wei

Gan

Qiu

Lan

Chen

Wang

Lin

Chen

and Mo

, Comprehensive investigation of aberrant microRNA profiling in bladder cancer tissues, Tumour Biol 37 (2016), 12555–69.

14.

Neerincx

Sie

D.L.

van de Wiel

M.A.

van Grieken

N.C.

Burggraaf

J.D.

Dekker

Eijk

P.P.

Ylstra

Verhoef

Meijer

G.A.

Buffart

T.E.

and Verheul

H.M.

, MiR expression profiles of paired primary colorectal cancer and metastases by next-generation sequencing, Oncogenesis 4 (2015), e170.

15.

Wang

Ding

Nan

Wang

Yan

Zhang

Sun

Zhu

Dong

and Yu

, Investigation of the roles of exosomes in colorectal cancer liver metastasis, Oncol Rep 33 (2015), 2445–53.

16.

Tanaka

Kaida

Yokoi

Ishii

Nishizawa

Kawamata

Katoh

Sato

Nakamura

Watanabe

and Yamashita

, Critical relevance of genomic gains of PRL-3/EGFR/c-myc pathway genes in liver metastasis of colorectal cancer, Oncol Lett 17 (2019), 1257–66.

17.

Duffy

M.J.

, Personalized treatment for patients with colorectal cancer: role of biomarkers, Biomark Med 9 (2015), 337–47.

18.

Deng

and Chen

, Increased Expression of miR-29a and Its Prognostic Significance in Patients with Cholangiocarcinoma, Oncol Res Treat 40 (2017), 128–32.

19.

Feng

Wang

and Kang

, Diagnostic significance of miR-210 as a potential tumor biomarker of human cancer detection: an updated pooled analysis of 30 articles, Onco Targets Ther 12 (2019), 479–93.

20.

Zhou

and Ma

, miR-378 serves as a prognostic biomarker in cholangiocarcinoma and promotes tumor proliferation, migration, and invasion, Cancer Biomark (2018).

21.

Cheng

Pang

Zhao

Chen

Wang

Qin

Han

Shi

and Fu

, High expression of MiR-98 is a good prognostic factor in acute myeloid leukemia patients treated with chemotherapy alone, J Cancer 10 (2019), 178–85.

22.

Liu

Sun

and Kan

, miR-935 Promotes Liver Cancer Cell Proliferation and Migration by Targeting SOX7, Oncol Res 25 (2017), 427–35.

23.

Yang

Cui

Ding

Yang

Liu

Dai

and Chen

, miR-935 promotes gastric cancer cell proliferation by targeting SOX7, Biomed Pharmacother 79 (2016), 153–8.

24.

Muhammad

Tang

Wei

Zhang

Wang

Muhammad

B.U.

Kaur

Kamchedalova

Gang

Jiang

Liu

and Wang

, miRNA-30d serves a critical function in colorectal cancer initiation, progression and invasion via directly targeting the GNA13 gene, Exp Ther Med 17 (2019), 260–72.

25.

Liu

J.L.

J.J.

Xiao

Cui

P.L.

Qiao

Z.G.

and Zhang

X.L.

, MiR-144 Inhibits Tumor Growth and Metastasis in Osteosarcoma via dual-suppressing RhoA/ROCK1 Signaling Pathway, Mol Pharmacol (2019).

26.

Xing

Wang

and Zhou

, The Expression of MicroRNA-598 Inhibits Ovarian Cancer Cell Proliferation and Metastasis by Targeting URI, Mol Ther Oncolytics 12 (2019), 9–15.

27.

Wang

and Zhou

, microRNA-769 is downregulated in colorectal cancer and inhibits cancer progression by directly targeting cyclin-dependent kinase 1, Onco Targets Ther 11 (2018), 9013–25.

28.

Liu

Yang

Cheng

Chen

Cui

Sun

You

Liu

Sun

Wang

Zhang

and Li

, miR-19a promotes colorectal cancer proliferation and migration by targeting TIA1, Mol Cancer 16 (2017), 53.

29.

Wang

Huang

and Chen

, MicroRNA-940 Targets INPP4A or GSK3beta and Activates the Wnt/beta-Catenin Pathway to Regulate the Malignant Behavior of Bladder Cancer Cells, Oncol Res 26 (2018), 145–55.

30.

Zhang

Tian

Yan

and Xu

, MicroRNA-4286 promotes esophageal carcinoma development by targeting INPP4A to evoke the JAK2/STAT3 pathway activation, Pharmazie 73 (2018), 342–48.

MicroRNA-935 acts as a prognostic marker and promotes cell proliferation,migration,and invasion in colorectal cancer

Abstract

Keywords

Table 1 Correlation between miR-935 expression levels and clinical features in CRC patients

2. Materials and methods

2.1 CRC patients and tissue specimens

2.2 Cell lines and transfection

2.3 RNA extraction and real-time quantitative polymerase chain reaction (qRT-PCR)

2.4 Cell proliferation assay

2.5 Migration and invasion assays

2.7 Statistical analysis

3. Results

3.1 Expression of miR-935 in CRC tissues and cells

3.2 The association between miR-935 expression and clinical parameters of CRC patients

3.3 The prognostic value of miR-935 in CRC

Table 2 Multivariate Cox analysis of clinical parameters in relation to overall survival

Footnotes

Acknowledgments

Conflict of interest

References

Table 1
Correlation between miR-935 expression levels and clinical features in CRC patients

Table 2
Multivariate Cox analysis of clinical parameters in relation to overall survival