MiR-122 Promotes the Development of Colon Cancer by Targeting ALDOA In Vitro.

Non-coding RNAs, originally considered junk gene products, have taken center stage in view of their significant involvement in a spectrum of biological processes during human development, thereby offering novel therapeutic targets for improvement of treatment options. Accumulating evidence has demonstrated non-coding RNA dysfunction across various human cancers. In particular, microRNAs have emerged as key regulatory molecules in cancer biology. MicroRNAs are noninvasive, readily accessible biomarkers that can be effectively applied for diagnosis and prognosis of different tumor types, including colon cancer. In this study, we reanalyzed the available data with bioinformatics tools to identify differentially expressed microRNAs in colon cancer cells. The top 3 upregulated microRNAs (miR-10, miR-199, and miR-122) in colon cancer cells were further validated in tissues of clinical patients via reverse transcription-quantitative polymerase chain reaction. Our results showed that miR-122 significantly promotes the proliferation and invasion ability of SW480 and SW620 cells through inhibition of Aldolase, Fructose-Bisphosphate A (ALDOA) expression. We further summarized recent advances in our understanding of the functional relevance of microRNAs in cancer development and discussed the possible implications of specific microRNAs in colon cancer. This study extends our knowledge of microRNA involvement in colon cancer biology and presents novel candidates for the development of attractive therapeutic strategies.

Introduction

Cancer, a disease initiated by mutations in critical genes that result in uncontrolled proliferation and inappropriate survival of damaged cells, is a leading cause of death worldwide.[1] Early diagnosis should facilitate timely treatment and reduce recurrent events as well as morbidity and mortality.[2] Under normal conditions, cells exert several safeguarding measures to ensure that division, differentiation, and death take place at the expected times. Several regulatory factors function to switch on or off genes that guide cellular proliferation and differentiation.[3,4] Aberrant regulation of these tumor suppressors or oncogenes has been linked to a plethora of diseases, including cancer.[2,5] Therefore, the identification of noninvasive and highly sensitive biomarkers providing a detailed and accurate fingerprint of specific diseases remains an urgent clinical requirement. Such biomarkers provide useful information to diagnose and monitor disease conditions and effectively establish prognosis and appropriate treatments. Accumulating evidence has demonstrated the functional relevance of novel non-coding RNAs (ncRNAs) with altered expression patterns in tumorigenesis in multiple tumor types, supporting their utility in diagnostic and prognostic applications as well as therapeutic targets for cancer.[2,6]

About >75% of the human genome is transcribed (only 2% into protein-coding RNAs[7]), with increasing evidence of crucial roles of ncRNAs in disease development, particularly cancer.[8] Based on a length threshold, ncRNAs are categorized into 2 subgroups: short ncRNAs (<200 nucleotides), including piRNAs, microRNAs (miRNAs), and certain circRNAs, and long non-coding RNAs (lncRNAs >200 nucleotides).[9] Non-coding RNAs act as transcriptional or post-transcriptional regulators whose aberrant expression or impaired functional processes are highly correlated with cancerous pathophysiology.[10]

MicroRNAs, a conserved class of endogenous short ncRNAs 18 to 25 nucleotides in length that regulate target genes by inducing degradation or suppressing translation of target messenger RNAs (mRNAs) after transcription, have been actively investigated as anticancer therapy in recent years.[11] Unlike other ncRNA types, miRNAs interact with the Ago subfamily of the Argonaute family. Each miRNA has the potential to target multiple genes (∼500 for each miRNA family) that often involve similar pathways.[12,13]

Over 50% miRNAs are located in fragile sites and cancer-associated genomic regions, supporting crucial roles in tumorigenesis. MicroRNAs are reported to regulate numerous oncogenes or tumor suppressors and consequently modulate tumorigenesis.[14] For example, miR-34c-5p downregulates the amphiregulin (AREG)-epidermal growth factor receptor (EGFR)-extracellular signal-regulated kinase (ERK) pathway and subsequently hampers ovarian cancer stemness and drug resistance.[15] MicroRNA-151-3p inhibits migration of breast cancer cells by targeting TWIST1,[16] while miR-106b and miR-93 promote tumor cell progression by targeting Phosphatase And Tensin Homolog (PTEN) and subsequently activating the PI3K/Akt pathway in breast cancer.[17]

Colon cancer is one of the leading causes of cancer-induced mortality worldwide, with about 1 million new cases diagnosed annually.[18] Increasing age, male gender, and alcohol have been identified as risk factors for colorectal cancer development. Colon cancer is triggered by mutations in tumor suppressor or DNA repair genes.[19] A number of miRNAs with essential roles in colon cancer progression have been identified to date. For example, miR-193a-3p promotes carcinogenesis via upregulating IL17RD expression. Knockdown of miR-193b suppresses colon cancer cell growth via activation of transforming growth factor-β (TGF-β) and the small mothers against decapentaplegic (SMAD)3 pathway.[20]

Although numerous miRNA regulators of colon cancer have been identified, effective clinical applications are yet to be developed. Among the deaths caused by colon cancer, more than 50% cases showed evidence of liver metastasis, one of the major causes of mortality.[21] However, the underlying molecular mechanisms that mediate colorectal cancer metastasis to the liver have not been systemically characterized to date.

To further elucidate the roles of miRNAs in colon cancer development, bioinformatics tools were applied to analyze the miRNA-seq data from published data sets to obtain differentially expressed miRNAs between colon cancer and paired adjacent normal tissues. Along with a number of other miRNAs, miR-122, miR-10, and miR-199 were differentially expressed in colon cancer tissue. Comparison of expression patterns between in situ and liver metastatic colon cancer revealed higher expression of miR-122 in liver metastatic colon cancer samples. Among the predicted targets of miR-122, we identified ALDOA that participates in colon cancer metastasis. Our data suggest that miR-122 serves as an ncRNA regulator of colon cancer development by targeting ALDOA and potentially other cancer-related genes.

Method

Clinical Samples

Three adults with colon cancer voluntarily participated in our study. Tumor tissues were derived from initial surgery without either preoperative chemotherapy or radiotherapy at the hospital. Tissues were collected immediately after surgical removal and snap-frozen in liquid nitrogen until further use.

Identification of Differentially Expressed miRNAs

We performed miRNA microarray analysis using the interactive web tool, GEO2R, with the available GEO data (Accession: GSE48267). GEO2R was an interactive web tool that allows users to compare 2 or more groups of samples in a GEO Series in order to identify genes that are differentially expressed across experimental conditions (https://www.ncbi.nlm.nih.gov/geo/info/geo2r.html). All the 122 samples downloaded were classified into 2 groups, including colon cancer and adjacent normal colon tissue, which were used for subsequent analysis of miRNA quantification.[22] Raw probe-level intensity data were processed according to the procedure of Li et al.[22] After obtaining normalized miRNA, we used default parameters of the significance analysis of microarrays method[23] to perform differential miRNA expression analysis. Differentially expressed miRNAs were identified based on a significant threshold P value <.05.

Cell Culture, Transfection, and Dual-luciferase Reporter Assays

SW480 and SW620 cells were cultured in high-glucose Dulbecco’s modified Eagle’s medium (DMEM; HyClone, Logan, Utah) with 10% (v/v) fetal bovine serum (FBS; Gibco, Grand Island, New York) at 5% CO2 and 37°C. MicroRNA-122 mimic and inhibitor (complementary sequences of miR-122) and corresponding negative control (random sequences) were purchased from GenePharma (Shanghai, China). Cells were transfected with the miR-122 mimic, negative control (NC), miR-122 inhibitor or inhibitor NC (GenePharma) using Lipofectamine 2000 transfection reagent (Invitrogen, Carlsbad, California). Opti-MEM I Reduced Serum Medium (Gibco) was used to dilute Lipofectamine 2000 and nucleic acids. Cells were collected after 48 hours transfection for subsequent analyses. For luciferase assays, ALDOA-3′UTR or ALDOA-3′UTR-Mut was transfected at a concentration of 500 ng together with miR-122 mimics. After 24 hours transfection, luciferase activities were measured with a PerkinElmer 2030 Multilabel Reader (PerkinElmer, Waltham, Massachusetts). The detailed sequence information is presented in Supplementary Table S1.

Reverse Transcription-quantitative PCR

Total RNA was extracted from SW480 cells with the HP Total RNA Kit (Omega Bio-tek, Norcross, Georgia) and treated with DNase I (Thermo Scientific, Waltham, Massachusetts). The concentration and quality of RNA were assessed with a NanoDrop 2000 spectrophotometer (Thermo Scientific) and agarose gel electrophoresis. Total RNA was reverse-transcribed using the Revert Aid First Strand cDNA Synthesis Kit (Thermo Scientific). For miR-10, miR-199, and miR-122 quantification, stem-loop RT-PCR was executed with U6 small nuclear RNA as an internal control. All primers were designed based on miR-10, miR-199, and miR-122 sequences collected from miRBase release 21 (http://www.mirbase.org/). Primers used for qRT-PCR are shown in Supplementary Table S1. Reverse transcription-quantitative polymerase chain reaction (RT-qPCR) was performed in triplicate using iQ SYBR green Supermix (Bio-Rad, Hercules, California) and the LightCycler®480 (Roche Applied Science, Penzberg, Upper Bavaria, Germany).

Western Blot Analysis

Cellular protein lysates were generated using RIPA Lysis Buffer (Beyotime, Shanghai, China) and extracted 48 hours after transfection. Proteins were separated via sodium dodecyl sulfate–polyacrylamide gel electrophoresis and a Mini Trans-Blot Cell (Bio-Rad) used to transfer protein onto polyvinylidene fluoride membranes (Millipore, Billerica, Massachusetts). Primary antibodies specific for ALDOA (ab169554, 1:5000; Abcam, Cambridge, Massachusetts) and β-actin (ab8226, 1:2000; Abcam) were used for immunoblot experiments. Protein expression was detected using an Image Quant LAS4000 mini instrument (GE Healthcare Life Sciences, Piscataway, New Jersey).

MTT Assay

All cells were maintained in low serum, containing 2% FBS (Invitrogen) with starvation for 10 hours before the experiments were performed. An aliquot of cells (1 × 103 cells/well) was seeded in a 96-well plate with 2% FBS and serum-starved for 10 hours, transfected with corresponding vector or siRNA and cultured in normal medium. At 24, 48, 72, and 96 hours after transfection, MTT (3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyl-tetrazolium bromide) solution (5 mg/mL, 20 μL) was added to each well. After incubation for 4 hours, the medium was removed and 100 μL dimethyl sulfoxide was added to each well. The relative number of surviving cells was assessed by measuring the optical density of cell lysates at 570 nm. All assays were performed 5 times.

Cell Transwell Assay

The cell transwell assay was performed using 24-well transwell chambers and polycarbonate membranes with a pore size of 8 µm (Corning Incorporated, Corning, New York). Briefly, 5 × 104 transfected cells were seeded into the upper transwell chambers coated with Matrigel (BD Biosciences, Bedford, Massachusetts) in serum-free medium, and the lower chambers were filled with DMEM containing 10% FBS. After incubation at 37°C for 24 hours, noninvading cells on the upper membrane surface were removed with cotton swabs, whereas cells that migrated onto the lower surface were fixed with 100% methanol and stained with 0.1% crystal violet. Images of cells on the lower surface were obtained and quantified by counting from 5 random fields in each well at a magnification of 200× under a microscope (Olympus, szx16, Japan).

Statistical Analysis

Data were presented as means ± standard deviation from 3 independent experiments and analyzed via the Student t test to calculate the significant differences between cancer and normal groups using GraphPad Prism 5.0 software (San Diego, California). The results were considered statistically significant at *P < .05, **P < .01, ***P < .001, and ****P < .0001 (n.s. represents nonsignificant).

Results

Differentially Expressed miRNAs in Colon Cancer

Based on the available microarray data comparing the miRNA expression profiles between colon cancer and adjacent normal colon tissue,[22] we downloaded and reanalyzed data from the GEO database using GEO2R software to identify differentially expressed miRNAs that may serve as therapeutic targets for colon cancer (Table 1). Overall, 17 differentially expressed miRNAs were identified, with P < .05 as the threshold of statistical significance. Overall, we observed 11 miRNAs that were downregulated in cancer tissue (Table 1). Among the top 3 miRNAs, miR-122 was upregulated while the abundance of miR-10 and miR-199 declined to a significant extent (Table 1). To validate the results, the top 3 differentially expressed miRNAs, miR-10, miR-199, and miR-122 were selected for subsequent analyses.

Table 1.

List of Differentially Expressed miRNAs.

miRNA_ID	Log (Fold Change)	P Value
hsa-miR-199b-5p	−2.349	.000
hsa-miR-122	4.233	.001
hsa-miR-10b	−4.404	.002
hsa-let-7i	−0.927	.007
hsa-miR-885-5p	3.337	.009
hsa-miR-766	−2.296	.016
hsa-miR-143	−0.987	.018
hsa-miR-515-5p	2.133	.020
hsa-miR-145	−1.202	.022
hsa-miR-1253	1.829	.025
hsa-miR-144	−1.335	.026
hsa-miR-548h	2.715	0.032
hsa-miR-520e	−2.252	0.038
hsa-miR-1	−2.589	.039
hsa-miR-451	−1.801	.043
hsa-miR-589	1.398	.047
hsa-miR-544	−4.761	.049

The miR-199 is of considerable interest in cancer therapy based on its observed association with various tumor types.[24,25] This miRNA is reported to delay progression from chronic hepatitis to hepatocellular carcinoma (HCC). The miR199-a/b-5p suppresses its target gene, Rho-associated coiled-coil kinase 1 (ROCK1), leading to inhibition of ROCK1/MLC and PI3K/Akt signaling and subsequently, HCC metastasis.[25] In the present study, we observed downregulation of miRNA-199 in colon tumor tissue, highlighting its potential role as a promising biomarker for clinical diagnosis and prognosis of colon cancer.

The miR-10 family members are encoded in evolutionarily conserved loci within the homeobox (Hox) gene clusters and play crucial roles in cell proliferation and apoptosis.[26] Both miR-10a and miR-10b compromise human granulosa cell development during folliculogenesis by targeting brain derived neurotrophic factor (BDNF), leading to suppression of the TGF-β pathway.[17] Downregulation of miRNA-10 is further suggestive of specific roles in colon cancer development.

Previous studies have shown that miR-122 functions as a tumor suppressor and undergoes downregulation in several cancer types, such as HCC, bladder cancer, and breast cancer.[27-29] MicroRNA-122 is also reported to hamper glioma cell invasion via inactivation of the Wnt/b-catenin signaling pathway through targeting of WNT1.[30] Conversely, the significant upregulation of miRNA-122 observed in this study supports potential roles in tumorigenesis or progression of colon cancer (Figure 1).

Figure 1.

Relative expression of miR-122, miR-10, and miR-199. Reverse transcription-quantitative polymerase chain reaction (RT-qPCR) was performed with 3 replicates to determine the relative expression levels of miR-122 (A), miR-10 (B), and miR-199 (C) in colon and metastatic liver tissues. Three colon cancer and metastatic liver tissue samples were used for the experiment.

MicroRNA-122 Promotes Proliferation and Metastasis of Colon Cancer Cell Lines

Based on the above analysis, we further explored variations in these 3 miRNAs in colon cancer and metastatic cancer tissue in liver. To this end, tumor tissues were collected from colon and liver immediately after surgical removal. Reverse transcription-quantitative polymerase chain reaction assays revealed dramatic repression of miRNA-199 and miRNA-10 in metastasis liver tissue along with upregulation of miR-122 (Figure 1), indicating contributory effects of miRNA-122 to progression and metastasis of colon cancer. To examine this hypothesis, the functional relevance of miRNA-122 in the migration ability of SW480 colon cancer cells was explored with the aid of the cell transwell assay. Cell migration of SW480 was promoted upon overexpression of miR-122, compared to that of the control group, while inhibition of miR-122 expression reversed this trend (Figure 2A). Similar cell migration abilities were observed for SW620 cells under comparable experimental conditions (Figure 2B). These results clearly indicate that miR-122 stimulates the migration ability of colon cancer cells.

Figure 2.

Overexpression of miR-122 promotes migration and proliferation in SW480 and SW620 cells. (A) SW480 cells were transfected with miR-122 mimic, negative control (NC), miR-122 inhibitor or Inhibitor NC. The transwell chamber assay was performed to determine SW480 cell migration. (B) The same experiments as (A) were performed using SW620 cells. (C) Bar plot showing that miR-122 promotes colon cancer SW480 cell proliferation. (D) Bar plot showing that miR-122 promotes colon cancer SW620 cell proliferation. Three and 5 replicate experiments were performed for the transwell chamber and MTT assay, respectively.

To further explore the function of miR-122 in colon cancer, expression levels were regulated with the aid of an miR-122 mimic or inhibitor and the MTT assay subsequently performed to assess cell proliferation. After 24, 48, and 72 hours, increased miR-122 levels induced a significant increase in proliferation of SW480 cells while knockdown of miR-122 led to a significant decrease in proliferation. Meanwhile, no marked differences were evident between mimic NC and normal (NC) cells (Figure 2C). Analogous results were obtained for SW620 cells (Figure 2D), clearly supporting a stimulatory effect of miR-122 on proliferation of colon cancer cells.

Targets Analysis of MicroRNA-122

Prediction of the binding targets of miRNAs is a key step in exploring their biological functions in living cells. Accordingly, we applied TargetScan (version 7.2) software[31] to predict the potential target transcripts of miR-122 with default parameters. In total, 226 transcripts were determined as miR-122 targets (Supplementary Table S2). To further explore the function of miR-122, enriched Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analyses of target transcripts were conducted. GO analysis revealed that the majority of the target genes are involved in cell growth, including negative regulation of GTPase activity, regulation of cell differentiation, and multicellular organism growth and nucleus (cellular component). Interestingly, negative regulation of GTPase activity, which is important for cell migration and invasion, was similarly enriched among the GO terms (Figure 3A). Moreover, target genes of miR-122 were involved in pathways related to biosynthesis and metabolism, including metabolic pathways, gluconeogenesis, and the citrate cycle (tricarboxylic acid (TCA) cycle; Figure 3B).

Figure 3.

Functional analysis of miR-122 target genes. (A) The top 10 enriched GO terms for target genes of miR-122. (B) The top 10 enriched KEGG pathways for target genes of miR-122. The X-axis number represents the enriched P value of each GO term or KEGG pathway. The numbers of each bar represent the target or background gene number of each term or pathway.

Aldolase, fructose-bisphosphate A (ALDOA) was one of the predicted targets of miR-122 (Supplementary Table S2). Previous studies have shown that ALDOA colocalizes with proliferation cell nuclear antigen (PCNA) in the cytoplasm and is involved in cell growth pathways,[32] and disruption of this gene contributes to the progression of multiple cancer types.[33] Accordingly, ALDOA was selected as a candidate gene to further explore the function of miR-122. The dual-luciferase reporter system was applied to validate the interactions between miR-122 and ALDOA transcripts. Upon co-transfection of miR-122 mimics with a luciferase reporter vector containing ALDOA-3′UTR (ALDOA-3′UTR) into SW480 cells, luciferase activity was significantly suppressed (Figure 4A). In contrast, luciferase activity remained unchanged upon co-transfection of miR-122 mimics with ALDOA-3′UTR-Mut luciferase reporters (Figure 4A), implying physical interactions between miR-122 and ALDOA. Reverse transcription-quantitative polymerase chain reaction and western blot findings additionally revealed significant suppression of mRNA and protein expression of ALDOA following transfection of miR-122 mimics into SW480 cells (Figure 4B and C). Inhibition of miR-122 led to a significant increase in ALDOA mRNA and protein expression (Figure 4B and C).

Figure 4.

Identification of ALDOA as a direct target of miR-122 in SW480 cells. (A) Luciferase activity was analyzed 24 hours after co-transfection of SW480 cells with ALDOA-3′UTR or ALDOA-3′UTR-Mut and miR-122 mimic or mimic NC. (B) Endogenous ALDOA mRNA levels were detected 24 hours after transfection of SW480 cells with miR-122 mimic, mimic NC, miR-122 inhibitor, or Inhibitor NC. (C)Western blot was applied to detect endogenous ALDOA protein expression 48 hours after transfection of SW480 cells with miR-122 mimic, mimic NC, miR-122 inhibitor, or Inhibitor NC (*P < .05, **P < .01).

To explore the functions of ALDOA in colon cancer cells, its expression was suppressed using si-ALDOA and conversely increased by transferring pcALDOA into SW480 and SW620 cells, and the transwell assay was performed. Cell migration ability was enhanced upon repression of ALODA, compared to the control group (Figure 5A-B), which was reversed upon ALDOA overexpression (Figure 5A-B). Data from the MTT assay showed that overexpression of ALDOA induced significant repression of SW480 cell proliferation while inhibition of its expression led to a significant increase in proliferation (Figure 5C and D). These results suggest that miR-122 interacts directly with ALDOA, which may be involved in the progression of liver metastases of colon cancer and could therefore serve as an attractive therapeutic target for this cancer type.

Figure 5.

Downregulation of ALDOA promotes migration of SW480 and SW620 cells. (A) SW480 cells were transfected with pcALDOA, negative control (pcDNA3.1), si-ALDOA and si-NC, and the transwell chamber assay performed to determine SW480 cell migration. (B) The same experiments as (A) were performed using SW620 cells. (C) Bar plot showing that downregulation of ALDOA promotes colon cancer SW480 cell proliferation. (D) Bar plot showing that miR-122 promotes colon cancer SW620 cell proliferation. Three and 5 replicate experiments were performed for transwell and MTT assays, respectively. (E) Proposed working model of the involvement of miR-122, ALDOA and other potential mRNA targets in the development of colon cancer.

Discussion

Dysregulation of ncRNAs is involved in the progression of a variety of cancers. Detailed characterization of the ncRNA transcriptome and enhanced understanding of their features and therapeutic applications as well as the mechanisms by which they control tumorigenesis should facilitate the development of more effective treatment strategies. Emerging evidence has demonstrated that piRNAs, circRNAs, and miRNAs, which are promising biomarkers, display strong diagnostic and prognostic value in cancer treatment.

The well-established roles of miRNAs have aided in significant advancements in ncRNA research. Aberrant expression of miRNAs is strongly implicated in multiple cancers and specific miRNAs serve as crucial factors related to proliferation, apoptosis, and metastasis in colon cancer, making them valuable therapeutic agents.[34] The unique and conserved biogenesis of miRNAs mainly involves 3 steps: transcription, endonucleolytic processing in the nucleus and cytoplasm, and merging into the RNA-induced silencing complex.[35,36] Carcinogenesis is driven by molecular alterations leading to the activation of pro-survival, pro-proliferative, and metastatic signaling, such as WNT, TGF-β, and EGFR signaling pathways.[37] Overexpression of miR-195 hampers colorectal cancer cell proliferation through targeting fibroblast growth factor 2 and blocking Wnt/β-catenin pathways.[38] Upon deletion of miR-574-5p, cell proliferation, migration, and invasion are suppressed via targeting Qki6/7 to inactivate β-catenin/Wnt signaling in colorectal cancer.[39] MicroRNA-21, activated by the WNT signaling pathway, induces stemness through inactivation of TGFBR2 signaling in colon cancer.[40]

Cell invasion is the first step of cancer progression that involves tumor cells translocating from their primary site and metastasizing to distant organs.[41] Metastases are responsible for the majority of cancer deaths, in particular, metastatic liver disease. Therefore, early detection of liver metastasis is of utmost priority to allow radical surgery intervention and long-term survival.[42] Numerous studies have shown that miRNAs affect the expression of genes and pathways involved in cancer pathogenesis from initiation to metastasis.[43] MicroRNA-122 was initially reported as a highly expressed miRNA in liver that plays important roles in liver cancer development.[44,45] Moreover, miR-122 is associated with the development of different cancer types, including breast cancer,[46] renal cancer,[47] cholangiocarcinoma,[48] bile duct carcinoma cells,[49] and colorectal cancer.[50] Considering the altered expression of miR-122 in our experimental samples and the finding that its overexpression promotes SW480 cell invasion, we speculate that upregulation of miR-122 facilitates the metastatic progression of colon cancer in the liver through activating the WNT, TGF-β, or EGFR signaling pathway. One possible mechanism is that miR-122 promotes invasion of colon cancer by regulating the expression of ALDOA. Disruption of ALDOA plays a role in the progression of multiple cancer types, and aldolase has been validated as an independent clinical prognostic marker of human cancers.[51] Accordingly, we put forward the theory of a relationship between aberrant ALDOA expression and colon cancer invasion through miR-122 regulation. Functional analyses of all miR-122 targets showed that glycolysis/gluconeogenesis is the most enriched KEGG pathway, from which genes are extensively regulated in more than 20 cancer classes.[52] Our data imply that miR-122 promotes colon cancer invasion by regulating other target genes (Figure 5E).

Cancer progression can be prevented using several strategies including inhibition of oncogenic miRNAs, induction of tumor suppressor miRNAs, and decaying miRNA expression through epigenetic factors, such as promoter methylation.[43] The 3 miRNAs identified in this study (miR-122, miR-10, and miR-199) present potential attractive diagnostic and prognostic markers of liver metastasis of colon cancer and provide guidance for further development of miRNA-based therapeutic strategies against colon cancer.

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Supplemental Material, Supplementary_Table_S1 for MiR-122 Promotes the Development of Colon Cancer by Targeting ALDOA In Vitro by Hong Li, Xinhua Zhang, Zhao Jin, Tao Yin, Chuanyi Duan, Junwei Sun, Rui Xiong and Zilin Li in Technology in Cancer Research & Treatment

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Supplemental Material, Supplementary_Table_S2_miR-122_predicted_targets for MiR-122 Promotes the Development of Colon Cancer by Targeting ALDOA In Vitro by Hong Li, Xinhua Zhang, Zhao Jin, Tao Yin, Chuanyi Duan, Junwei Sun, Rui Xiong and Zilin Li in Technology in Cancer Research & Treatment