Integrating serum microRNAs and electronic health records improved the diagnosis of tuberculosis.

BACKGROUND: To verify the differential expression of miR-30c and miR-142-3p between tuberculosis patients and healthy controls and to investigate the performance of microRNA (miRNA) and subsequently models for the diagnosis of tuberculosis (TB). METHODS: We followed up 460 subjects suspected of TB, and finally enrolled 132 patients, including 60 TB patients, 24 non-TB disease controls (TB-DCs), and 48 healthy controls (HCs). The differential expression of miR-30c and miR-142-3p in serum samples of the TB patients, TB-DCs, and HCs were identified by reverse transcription-quantitative real-time PCR. Diagnostic models were developed by analyzing the characteristics of miRNA and electronic health records (EHRs). These models evaluated by the area under the curves (AUC) and calibration curves were presented as nomograms. RESULTS: There were differential expression of miR-30c and miR-142-3p between TB patients and HCs (p < 0.05). Individual miRNA has a limited diagnostic value for TB. However, diagnostic performance has been both significantly improved when we integrated miR-142-3p and ordinary EHRs to develop two models for the diagnosis of tuberculosis. The AUC of the model for distinguishing tuberculosis patients from healthy controls has increased from 0.75 (95% CI: 0.66-0.84) to 0.96 (95% CI: 0.92-0.99) and the model for distinguishing tuberculosis patients from non-TB disease controls has increased from 0.67 (95% CI: 0.55-0.79) to 0.94 (95% CI: 0.89-0.99). CONCLUSIONS: Integrating serum miR-142-3p and EHRs is a good strategy for improving TB diagnosis.

LncRNA NR2F2-AS1 promotes tumourigenesis through modulating BMI1 expression by targeting miR-320b in non-small cell lung cancer.

Recently, long noncoding RNAs (lncRNAs) are attracting wide attention in the field of cancer research because of its important role in cancer diagnosis and prognosis. But studies on the biological effects and relevant mechanisms of lncRNAs in non-small cell lung cancer (NSCLC) remain few and need to be enriched. Our study discussed the expression and biological effects of LncRNA NR2F2-AS1, and further explored its possible molecular mechanisms. As a result, elevated expression of NR2F2-AS1 was detected in NSCLC tissues and cells and was remarkably associated with the tumor, node, metastasis (TNM) stage and the status of lymphatic metastasis of patients. Down-regulated NR2F2-AS1 contributed to the promotion of cell apoptosis and the inhibition of cell proliferation and invasion in A549 and SPC-A-1 cells in vivo and vitro. Through bioinformatics analysis, NR2F2-AS1 functions as a ceRNA directly binding to miR-320b, BMI1 was a direct target of miR-320b. Combined with the following cellular experiments, the data showed that NR2F2-AS1 may influence the NSCLC cell proliferation, invasion and apoptosis through regulating miR-320b targeting BMI1.

Serum miR-146a, miR-155, and miR-210 as potential markers of Graves' disease.

BACKGROUND: Previous studies have demonstrated that dysfunctional regulatory T cells (Tregs) may be associated with Graves' disease (GD). In this study, we evaluated four serum Treg-associated miRNAs (miR-210, miR-182, miR-155, and miR-146a) expressions and assessed the potential of serum miRNAs as biomarkers of GD. METHODS: Foxp3 and serum miRNAs expressions both in GD patients and healthy controls were measured by RT-PCR. RESULTS: Serum miR-210 in GD patients was significantly higher than that of healthy controls (2.64-fold, P<.001); in contrast, miR-155 and miR-146a were lower (P<.001 and P=.008). No significant difference was found in miR-182. ROC curve analysis indicated that miR-210, miR-155, and miR-146a with the area under ROC (AUC) of 0.803 (70.0% sensitivity and 83.1% specificity), 0.796 (76.3% sensitivity and 76.9% specificity), and 0.736 (68.8% sensitivity and 73.8% specificity), respectively, could differentiate GD patients from healthy controls. Combination of three miRNAs yielded an AUC of 0.976 (91.3% sensitivity and 93.8% specificity) with 92.41% diagnostic efficiency. In addition, serum miR-210 and miR-155 in GD were associated with the extent of goiter. Three miRNAs levels were different by gender. Besides, serum miR-210 was positively correlated with free thyroxine (FT4) and thyrotrophin receptor antibody (TRAb) level. CONCLUSION: The serum levels of miR-210, miR-155, and miR-146a may be potential new markers for the diagnosis of GD and play important roles in GD pathogenesis.

RBM39 is a potential prognostic biomarker with functional significance in hepatocellular carcinoma.

Background: RNA-binding motif protein 39 (RBM39) is a well-known RNA-binding protein involved in tumorigenesis; however, its role in hepatocellular carcinoma (HCC) remains unclear. The aim of this study was to investigate the role of RBM39 in HCC. Methods: The Cancer Genome Atlas (TCGA) and Gene Expression Omnibus (GEO) databases were used to analyze the differential expression of RBM39 in HCC and normal tissues. The prognostic and diagnostic value of RBM39 in HCC was accessed by Kaplan-Meier analysis, Cox regression, and receiver operating characteristic (ROC) curve analyses. Quantitative real-time polymerase chain reaction (qRT-PCR) and immunohistochemistry were used to validate the mRNA and protein expression of RBM39 in HCC. Moreover, gene set enrichment analysis (GSEA) was performed to identify key pathways related to RBM39. The correlation between RBM39 expression and immune cell infiltration was evaluated using a single-sample gene set enrichment analysis (ssGSEA). CCK8 and wound healing assays were performed to investigate the proliferation and migration abilities of HCC cells with RBM39 knockdown. Results: RBM39 expression was upregulated in the HCC tissues. High RBM39 expression was significantly associated with advanced T stage, histological grade, and pathological stage and predicted poor overall survival (OS), disease-specific survival (DSS), and progress-free interval (PFI) in HCC patients. The upregulation of RBM39 expression was an independent prognostic factor for OS. Moreover, GSEA enrichment analysis indicated that RBM39 was functionally involved in pathways associated with the cell cycle, DNA replication, the p53 signaling pathway, and primary immunodeficiency. RBM39 expression was associated with infiltration of Th2 cells and dendritic cells (DC). RBM39 knockdown significantly inhibited the proliferation and migration of HCC cells. Conclusions: These findings suggest that high RBM39 expression is associated with poor prognosis and promotes HCC cell proliferation and migration. Based on these results, RBM39 is a promising prognostic biomarker with functional significance for HCC.

LncRNA AK142643 promotes hepatic lipid accumulation by upregulating CD36 via interacting with IGF2BP2.

Excessive lipid accumulation in hepatocytes is a defining feature of non-alcoholic fatty liver disease (NAFLD), a condition that is becoming increasingly prevalent worldwide. While long non-coding RNAs (LncRNAs) have been implicated in hepatic lipid metabolism, the precise regulatory mechanisms they employ remain poorly understood. In this study, we investigate the role of AK142643, a previously uncharacterized LncRNA, in hepatic lipid metabolism and the development of NAFLD. Our results demonstrate that AK142643 is upregulated in the livers of ob/ob and high fat diet (HFD)-fed mice, and that it promotes hepatic lipid accumulation both in vivo and in vitro. Furthermore, we reveal that AK142643 acts through the insulin-like growth factor 2 mRNA binding protein 2 (IGF2BP2) to enhance the expression of fatty acid translocase (FAT)/CD36, a key regulator of lipid metabolism. Specifically, AK142643 facilitates the binding of IGF2BP2 to CD36 mRNA, thereby increasing its stability and promoting its expression. Taken together, these findings shed new light on the complex interplay between LncRNAs and hepatic lipid metabolism, and provide insights into the mechanisms underlying the development of NAFLD.

Gene Signatures and Prognostic Values of m6A Regulators in Hepatocellular Carcinoma.

N6-methyladenosine (m6A) is the most abundant mRNA modification in mammals and has been implicated in various biological processes. However, its role in hepatocellular carcinoma (HCC) remains largely unknown. In this study, we investigated the alterations of 19 main m6A regulatory genes in HCC and their association with clinicopathological features, including survival. The mutation, copy number variation (CNV) and clinical data of HCC patients were retrieved from The Cancer Genome Atlas (TCGA) database. We found that the m6A regulators had high frequent alterations in HCC. The alterations of m6A regulators were significantly associated with clinicopathological features as well as TP53 alteration. Patients with any mutation of the m6A regulatory genes had worse overall survival (OS) and disease free survival (DFS). Deletion of METTL16 or ALKBH5 predicted poor OS and DFS of HCC patients. Moreover, deletion of METTL16 was an independent risk factor for DFS. Low METT16 expression was association with activation of multiple metabolic pathways in HCC. Finally, by RT-PCR, we confirmed that METTL16 was downregulated in HCC, and that lower METTL16 expression was associated with poor OS. In conclusion, we reported a significant association between alterations of m6A regulators and clinicopathological features, and highlighted the importance of METTL16 among the 19 m6A regulators in HCC pathogenesis. These findings will provide new insights into the role of m6A modification in HCC.

Expression levels and prognostic values of annexins in liver cancer.

Annexins are a superfamily of calcium-dependent phospholipid-binding proteins that are implicated in a wide range of biological processes. The annexin superfamily comprises 13 members in humans (ANXAs), the majority of which are frequently dysregulated in cancer. However, the expression patterns and prognostic values of ANXAs in liver cancer are currently largely unknown. The present study aimed to analyze the expression levels of ANXAs and survival data in patients with liver cancer from the Oncomine, GEPIA, Kaplan-Meier plotter and cBioPortal for Cancer Genomics databases. The results demonstrated that ANXA1, A2, A3, A4 and A5 were upregulated, whereas ANXA10 was downregulated in liver cancer compared with normal liver tissues. The expression of ANXA10 was associated with pathological stage. High expression levels of ANXA2 and A5 were significantly associated with poor overall survival (OS) rate whereas ANXA7 and A10 were associated with increased OS. The prognostic values of ANXAs in liver cancer were determined based on sex and clinical stage, which revealed that ANXA2, A5, A7 and A10 were associated with OS in male, but not in female patients. In addition, the potential biological functions of ANXAs were identified by Gene Ontology functional annotation and Kyoto Encyclopedia of Genes Genomes pathway analysis; the results demonstrated that ANXAs may serve a role in liver cancer through the neuroactive ligand-receptor interaction pathway. In conclusion, the results of the present study suggested that ANXA1, A2, A3, A4, A5 and A10 may be potential therapeutic targets for liver cancer treatment, and that ANXA2, A5, A7 and A10 may be potential prognostic biomarkers of liver cancer.

Prognostic role of long non-coding RNA HNF1A-AS1 in Chinese cancer patients: a meta-analysis.

BACKGROUND: Long non-coding RNAs (LncRNAs) play important roles in tumorigenesis and progression. Recent studies have demonstrated that LncRNA HNF1A antisense RNA 1 (HNF1A-AS1) is aberrantly expressed in several types of cancers and is associated with poor outcomes. This meta-analysis was conducted to investigate the relationship between HNF1A-AS1 expression and clinical outcomes in cancer patients. METHODS: We searched PubMed, Embase, Web of Science, China National Knowledge Infrastructure (CNKI), and Wan Fang databases (updated until December 31, 2017) for literature. A total of eight studies with 789 cancer patients were finally included in the present meta-analysis. RESULTS: The results showed that high expression of HNF1A-AS1 significantly predicted poor overall survival (HR=3.10, 95% CI: 1.58-6.11, P=0.001), which was further validated using The Cancer Genome Atlas (TCGA) dataset. Moreover, high HNF1A-AS1 expression was also associated with advanced TNM stage (OR=3.32, 95% CI: 2.28-4.83, P<0.001), lymph node metastasis (OR=3.08, 95% CI: 1.95-4.85, P<0.001), and distant metastasis (OR=5.53, 95% CI: 1.94-15.77, P=0.001). CONCLUSION: Our results suggested that elevated HNF1A-AS1 was associated with poor clinical outcomes and might serve as a potential prognostic biomarker of cancer.

Functional miR-146a, miR-149, miR-196a2 and miR-499 polymorphisms and the susceptibility to hepatocellular carcinoma: An updated meta-analysis.

OBJECTIVE: Single nucleotide polymorphisms of miRNAs play important roles in the pathogenesis of hepatocellular carcinoma (HCC). To evaluate the association between four common miRNAs (miR-146a rs2910164; miR-149 rs2292832; miR-196a2 rs11614913 and miR-499 rs3746444) and HCC risk, an updated meta-analysis was performed. METHODS: 32 studies including 12,405 HCC cases and 15,056 controls were used for this meta-analysis. There were 22 studies with 7894 cases and 10,221 controls for miR-146a, 9 studies with 2684 HCC cases and 3464 controls for miR-149, 17 studies with 6937 cases and 8217 controls for miR-196a2 and 16 studies with 4158 cases and 5264 controls for miR-499. Odds radios (ORs) and 95% confidence intervals (CIs) were used to assess the HCC risk. RESULTS: Meta-analysis showed that miR-146a was associated with HCC risk under the heterozygote model (OR=1.10, 95%CI=1.03-1.17, P=0.007), whereas no association was found in Caucasian using all genetic models. For miR-196a2 polymorphism, an increased risk of HCC was observed based on four models (C vs T: OR=1.15, 95%CI=1.05-1.26, P=0.003; CC vs TT: OR=1.35, 95%CI=1.12-1.63, P=0.002; CC+CT vs TT: OR=1.20, 95%CI=1.04-1.37, P=0.01 and CC vs CT+TT: OR=1.23, 95%CI=1.06-1.42, P=0.006). Association of miR-499 with HCC risk was only detected in the subgroup of studies which did not use polymerase chain reaction and restriction fragment length polymorphism (PCR-RFLP) under the allelic, heterozygote and dominant models. However, negative results were obtained for the association of miR-149 and HCC susceptibility. CONCLUSION: Our results suggest that miR-146a and miR-196a2 polymorphisms are associated with increased risk of HCC, especially in Asian.

miR-455-5p functions as a potential oncogene by targeting galectin-9 in colon cancer.

Although there is evidence that galectin-9 is a critical factor in health and disease, the upstream regulatory microRNA (miRNA or miR) of the protein remains poorly defined. miR-455-5p is characterized as a tumor-associated miRNA in cancer research. However, the actual role of miR-455-5p with respect to inhibiting or promoting tumorigenesis in colon cancer is unclear. The present study aimed to investigate the expression, role and target regulation association of galectin-9 and miR-455-5p in colon cancer. Western blot analysis and reverse transcription-quantitative polymerase chain reaction were used for the detection of the expression levels of galectin-9 and miRNAs. Cell Counting kit-8 test was used for the evaluation of cell proliferation, while flow cytometry was used for cell apoptosis analysis. A potential interaction between galectin-9 and miR-455-5p was predicted by target prediction programs and confirmed by luciferase assay and transfection with miRNA mimics. The present study revealed that elevated expression of galectin-9 and miR-455-5p in colon cancer was associated with HT29 cell proliferation and apoptosis. Furthermore, the present study demonstrated that miR-455-5p reduced galectin-9 expression by directly targeting its 3'-untranslated region. These data suggest that miR-455-5p functions as a potential oncogene in colon cancer by targeting galectin-9.

Role of PALB2 Polymorphisms with Regard to Susceptibility to Female Breast Cancer Risk in the Chinese Population.

AIMS: PALB2 (partner and localizer of BRCA2) is a nuclear partner of BRCA2 and promotes its localization and stability in the nucleus, allowing it to function in DNA repair and at the S-phase checkpoint. It has been hypothesized that polymorphisms in the PALB2 gene may be associated with tumorigenesis, particularly with respect to susceptibility to breast cancer. METHODS: To assess the association of polymorphisms in the PALB2 gene with breast cancer risk in a Chinese female population a total of 351 female breast cancer patients and 360 age-, gender-matched tumor-free individuals were enrolled in this case-control study. The genotypes of five PALB2 gene polymorphic sites (rs120963, rs16940342, rs249954, rs447529, and rs249935) were characterized by using the Sequenom MassARRAY platform. RESULTS: The data showed that the genotypes rs249954 CT (adjusted odds ratio [OR] = 1.52; 95% confidence interval [95% CI], 1.10-2.09), TT (adjusted OR = 2.36; 95% CI, 1.39-4.02), and CT/TT (adjusted OR = 1.65; 95% CI, 1.22-2.24) were associated with increased risk of breast cancer, respectively, relative to the CC genotype. Similarly, the rs120963 TC (adjusted OR = 1.89; 95% CI, 1.38-2.59), CC (adjusted OR = 3.88; 95% CI, 1.75-8.60), and TC/CC (adjusted OR = 2.05; 95% CI, 1.51-2.78) genotypes were associated with increased risk of breast cancer, respectively, relative to the TT genotype. Additionally, a weakly significant association was observed between G allele (AG/GG genotype) carriers of the rs249935 SNP in the sub-group of T1-2 (Adjusted OR = 1.43; 95% CI, 1.03-10.84) and negative lymphatic involvement (Adjusted OR = 3.23; 95% CI, 0.97-10.84) and risk of breast cancer. CONCLUSIONS: This case-control study provided evidence that rs120963 and rs249954 of the PALB2 gene are associated with increased breast cancer risk, and that the association of rs249935 with breast cancer risk may be modified by the tumor pathological characteristics.

Downregulation of miR-377 contributes to IRX3 deregulation in hepatocellular carcinoma.

Iroquois homeobox (IRX) gene family, which plays essential roles in embryonic development, has recently been reported to be involved in tumor progression. However, the association of IRX3, a member of the IRX family, with hepatocellular carcinoma (HCC) has not previously been studied. In the present study, we found that IRX3 was upregulated in HCC cell lines (HepG2 and SMMC7721). We investigated the regulatory mechanism of IRX3 in HCC cells. Western blot and luciferase reporter assays identified that IRX3 is a direct target of miR-377. In addition, miR-377 was downregulated in HepG2 and SMMC7721 cell lines, and overexpression of miR-377 inhibited HepG2 cell proliferation, migration and invasion. Moreover, miR-377 restoration significantly abrogated IRX3-induced proliferation, migration and invasion of SMMC7721 cells. These findings demonstrate the tumor-promoting potential of IRX3, and that downregulation of miR-377 may contribute to the upregulation of IRX3 in HCC. The present study provides insights into HCC progression and a novel potential therapeutic target of HCC treatment.

p-CREB-1 promotes hepatic fibrosis through the transactivation of transforming growth factor-ß1 expression in rats.

Phosphorylated cAMP-responsive element binding protein-1 (p-CREB-1) is an important transcription factor which has been reported to be implicated in fibrogenesis. However, the association between p-CREB-1 and transforming growth factor-ß1 (TGF-ß1)-mediated liver fibrogenesis remains poorly understood. In the present study, exogenous TGF-ß1 recombinant protein was used to activate hepatic stellate cells (HSCs), and we established a rat model of tetrachloromethane (CCl4)­induced liver fibrosis. Loss- and gain-of-function studies were performed to examine the role of p-CREB-1 in liver fibrogenesis, and the detailed mechanism responsible for these effects was further explored using chromatin immunoprecipitation and luciferase reporter gene assays. We found that p-CREB-1 expression was significantly upregulated in a rat model of hepatic fibrosis. We also demonstrated that p-CREB-1 increased TGF-ß1 expression and auto­induction in HSCs, through directly binding to the CRE site within the TGF-ß1 promoter in order to enhance its transcriptional activity. Moreover, lentivirus-mediated CREB-1 overexpression promoted hepatic fibrogenesis in rats. These findings suggest that p-CREB-1 may function as a potent profibrogenic factor through the transactivation of TGF-ß1 expression in liver fibrosis.

Reciprocal negative feedback loop between EZH2 and miR-101-1 contributes to miR-101 deregulation in hepatocellular carcinoma.

Although the tumor suppressive role of miR-101 is well documented in hepatocellular carcinoma (HCC), how the expression of miR-101 itself is regulated remains elusive. In the present study, we demonstrated that the miR-101 precursor pre-miR-101-1 could be regulated by an important epigenetic regulator, the enhancer of zeste homolog 2 (EZH2). Reporter gene assays revealed that ectopic expression of EZH2 inhibited the transcriptional activities of miR-101-1 promoter. Subsequent analyses revealed that miR-101-1 directly represses the expression of EZH2, and miR-101-1 and EZH2 form a reciprocal negative feedback loop as indicated by the fact that ectopic mature miR-101 could induce endogenous pre-miR-101-1 expression. This mature miR-101-induced pre-miR-101 expression was specific to pre-miR-101-1 and depended on EZH2 activities. Moreover, our results also demonstrated that similar antitumor effects can be achieved either by ectopic miR-101 or EZH2 silencing in HCC cells. These findings show that elevated EZH2 contributes to miR-101 deregulation in HCC and highlight the coordinated role of miR-101 and EZH2 in hepatocarcinogenesis.

A double-negative feedback loop between EZH2 and miR-26a regulates tumor cell growth in hepatocellular carcinoma.

Accumulating evidence demonstrates the important roles of microRNAs (miRNAs) in tumor development and progression. miR-26a has been reported to be downregulated in several types of cancers including hepatocellular carcinoma, but the underlying mechanism of how miR-26a is repressed remains largely unknown. In the present study, we performed western blot analysis, qRT-PCR, luciferase reporter assay and chromatin immunoprecipitation assay to investigate the relationship between miR-26a and the enhancer of zest homologue 2 (EZH2). CCK-8 assay and colony formation assay were carried out to explore the effect of miR-26a on HCC cells proliferation. We demonstrated that miR-26a was epigenetically repressed by EZH2-mediated H3K27 trimethylation within the miR-26a promoter. Moreover, we confirmed that EZH2 was also a direct target of miR-26a in HCC cells, thus, creating a double-negative feedback loop. Furthermore, miR-26a restoration increased the expressions of its host genes (CTDSPL and CTDSP2). Overexpression of EZH2 abrogated miR-26a induction of CTDSPL and CTDSP2. Restoring the balance of the double-negative feedback loop by miR-26a overpression or EZH2 silence significantly inhibited HCC cell growth. Overexpression of EZH2 rescued the growth inhibition effect of miR-26a. These findings suggest that an imbalanced double-negative feedback loop between EZH2 and miR-26a exists in HCC cells, which contributes to miR-26a deregulation and regulates tumor cells proliferation.

Serum miR-21, miR-26a and miR-101 as potential biomarkers of hepatocellular carcinoma.

AIM: This study aimed to investigate the expressions of serum miR-21, miR-26a and miR-101 in hepatocellular carcinoma (HCC) and their diagnostic value. METHODS: Serum levels of miR-21, miR-26a and miR-101 were detected by quantitative real-time polymerase chain reaction (qRT-PCR) in 52 HCC patients, 42 chronic hepatitis (CH) patients and 43 healthy controls. ROC curve analysis was performed to evaluate the diagnostic value. Clinical parameters were collected. RESULTS: Serum level of miR-21 was higher while miR-26a and miR-101 were significantly lower in HCC patients than those in healthy controls (P<0.05, P<0.001 and P<0.05, respectively). Serum levels of miR-26a and miR-101 were significantly lower in HCC patients than those in CH patients (P<0.001 and P<0.05). ROC curve analyses revealed that miR-21, miR-26a and miR-101 could differentiate HCC patients from healthy controls, the area under ROC curve (AUC) were 0.621 (67.4% sensitivity and 55.8% specificity), 0.754 (51.9% sensitivity and 95.2% specificity) and 0.631 (47.1% sensitivity and 81% specificity), respectively. Combination of miRNAs and alpha-fetoprotein (AFP) yielded an AUC of 0.914 with 87.0% sensitivity and 78.0% specificity. miR-26a and miR-101 had diagnostic potential for differentiating HCC from CH with AUC of 0.762 (75% sensitivity and 70% specificity) and 0.623 (54.9% sensitivity and 76.9% specificity). Combination of miR-26a, miR-101 and AFP yielded an improved AUC than AFP alone (0.854 vs. 0.683). Notably, miR-26a could differentiate small tumors HCC (≤3cm) from CH with an AUC of 0.753 (80% sensitivity and 62.5% specificity). CONCLUSIONS: Serum miR-21, miR-26a and miR-101 are deregulated in HCC and can serve as potential biomarkers. Combination of these miRNAs and AFP provide a better detection than AFP alone. Serum miR-26a is a promising biomarker for early detection of HCC.

microRNA-22 downregulation of galectin-9 influences lymphocyte apoptosis and tumor cell proliferation in liver cancer.

Galectin-9 (Gal-9) plays an important role in both the immune response and tumor progression, while microRNAs act as tumor regulators to mediate tumorigenesis. However, the underlying molecular mechanisms remain unknown. In the present study, we investigated the relationship between Gal-9 and microRNA-mediated regulation in liver cancer. We examined Gal-9 expression using qRT-PCR and western blot analysis and found that it was markedly upregulated in human liver cancer cells compared with the level in normal hepatocytes. We co-cultured peripheral blood mononuclear cells (PBMCs) and tumor cells and observed that Gal-9 induced lymphocyte apoptosis and tumor cell immune escape using flow cytometric analysis and WST-1 assay. We found that miR-22 was downregulated in liver cancer tissues and cell lines and confirmed that miR-22 directly targeted the Gal-9 3'UTR and negatively regulated Gal-9 expression by luciferase reporter assay and transfection of microRNA mimics. We also observed that the Gal-9/miR-22 axis may influence lymphocyte apoptosis and tumor cell proliferation. These studies contribute to a further understanding of the microRNA­mediated regulation of the Gal-9 pathway and elucidate novel therapeutic targets for liver cancer.