Jianying He

Overexpression of members of the microRNA-183 family is a risk factor for lung cancer: A case control study

BackgroundLung cancer is the leading cause of cancer-related deaths worldwide. Early detection is considered critical for lung cancer treatment. MicroRNAs (miRNAs) have shown promise as diagnostic and prognostic indicators. This study was to identify specific miRNAs with diagnostic and prognostic value for patients with lung cancer, and to explore the correlation between expression profiles of miRNAs and patient survival.MethodsGene expression of members of the miR-183 family (miR-96, miR-182, and miR-183) were examined in 70 paired samples from lung cancer patients (primary cancer and non-cancerous tissues and sera), as well as 44 serum samples from normal volunteers and lung cancer cell lines by quantitative real-time reverse transcription polymerase chain reaction (RT-qPCR). The correlation between the expression of miRNAs in tissues, sera, and patient overall survival were also examined by log-rank and Cox regression analysis.ResultsExpression levels of members of the miR-183 family in lung cancer tumor and sera were higher than that of their normal counterparts. The miR-96 expression in tumors was positively associated with its expression in sera. Log-rank and Cox regression analyses demonstrated that high expression of tumor and serum miRNAs of the miR-183 family were associated with overall poor survival in patients with lung cancer.ConclusionsOur results suggest that the expressions of miR-96, miR-182, and miR-183 in tumor and sera may be considered potential novel biomarkers for the diagnosis and prognosis of lung cancer.

Expression of miR-29c, miR-93, and miR-429 as Potential Biomarkers for Detection of Early Stage Non-Small Lung Cancer

Background Altered expression of miRNA expression contributes to human carcinogenesis. This study was designed to detect aberrant miRNA expressions as a potential biomarker for early detection and prognosis prediction of non-small cell lung cancer (NSCLC). Methods miRNA array was used to profile differentially expressed miRNAs and Taqman-based quantitative RT-PCR assays were used to analyze levels of miR-29c, miR-93, and miR-429 expression in NSCLC tissue samples, corresponding normal tissue samples, and serum samples from 70 NSCLC patients as well as in serum samples from 48 healthy controls. Results Levels of miR-29c and miR-93 expression were upregulated in NSCLC tissues, while serum levels of miR-29c were also upregulated, but levels of serum miR-429 were decreased in NSCLC. Moreover, the levels of miR-429 expression in NSCLC tissues were associated with those in serum samples. Receiver operating characteristic (ROC) curve analysis showed that at the optimal cut-off point, the areas under the ROC curve for serum levels of miR-29c and miR-429 were 0.723 and 0.727, respectively, levels which are higher than that of carcinoma embryonic antigen (0.534) in diagnosis of stage I NSCLC. In addition, serum levels of miR-429 were associated with poor overall survival of NSCLC patients. Both univariate and multivariate analyses showed that serum miR-429 level was an independent prognostic predictor for NSCLC. Conclusions The results of the current study suggest that detection of serum miR-29c and miR-429 expression should be further evaluated as a novel, non-invasive biomarker for early stage NSCLC.

Increased Tim-3 expression in peripheral NK cells predicts a poorer prognosis and Tim-3 blockade improves NK cell-mediated cytotoxicity in human lung adenocarcinoma.

T cell immunoglobulin- and mucin-domain-containing molecule-3 (Tim-3) has been shown to play an important role in mediating NK-cell function in human diseases. However, the relationship between Tim-3 expression in natural killer (NK) cells and human lung adenocarcinoma remains unclear. We therefore investigated the expression of Tim-3 in NK cells and explored the effect of Tim-3 blockade on NK cell-mediated activity in human lung adenocarcinoma. Upregulated expression of Tim-3 on CD3-CD56+ cells (P<0.05) and CD3-CD56(dim) cells (P<0.05) of patients with lung adenocarcinoma was detected by flow cytometry. Moreover, Tim-3 expression in CD3-CD56+ NK cells was higher in patients with lung adenocarcinoma with lymph node metastasis (LNM) (P<0.05) or with tumor stage T3-T4 (P<0.05). Tim-3 expression in CD56(dim) NK-cell subset was higher in patients with tumor size ≥3cm (P<0.05), or LNM (P<0.05) or with tumor stage T3-T4 (P<0.05). Further analysis showed that higher expressions of Tim-3 on both CD3-CD56+ NK cells and CD56(dim) NK-cell subset were independently correlated with shorter overall survival of patients with lung adenocarcinoma (log-rank test, P=0.0418, 0.0406, respectively). Importantly, blockade of Tim-3 signaling with anti-Tim-3 antibodies resulted in the increased cytotoxicity and IFN-γ production of peripheral NK cells from patients with lung adenocarcinoma. Our data indicate that Tim-3 expression in NK cells can function as a prognostic biomarker in human lung adenocarcinoma and support that Tim-3 could be a new target for an immunotherapeutic strategy.

Diagnostic Value of Serum miR-182, miR-183, miR-210, and miR-126 Levels in Patients with Early-Stage Non-Small Cell Lung Cancer

Blood-circulating miRNAs could be useful as a biomarker to detect lung cancer early. We investigated the serum levels of four different miRNAs in patients with non-small cell lung cancer (NSCLC) and assessed their diagnostic value for NSCLC. Serum samples from 112 NSCLC patients and 104 controls (20 current smokers without lung cancer, 23 pneumonia patients, 21 gastric cancer patients, and 40 healthy controls) were subjected to Taqman probe-based quantitative reverse transcription–polymerase chain reaction (RT-PCR). The data showed that the serum levels of miR-182, miR-183, and miR-210 were significantly upregulated and that the miR-126 level was significantly downregulated in NSCLC patients, compared with the healthy controls. Further receiver operating characteristic (ROC) curve analysis revealed that the serum miR-182, miR-183, miR-210, or miR-126 level could serve as a diagnostic biomarker for NSCLC early detection, with a high sensitivity and specificity. The combination of these four miRNAs with carcinoembryonic antigen (CEA) further increased the diagnostic value, with an area under the curve (AUC) of 0.965 (sensitivity, 81.3%; specificity, 100.0%; and accuracy, 90.8%) using logistic regression model analysis. In addition, the relative levels of serum miR-182, miR-183, miR-210, and miR-126 could distinguish NSCLC or early-stage NSCLC from current tobacco smokers without lung cancer and pneumonia or gastric cancer patients with a high sensitivity and specificity. Data from the current study validated that the four serum miRNAs could serve as a tumor biomarker for NSCLC early diagnosis.

Prognostic Evaluation of CapG, Gelsolin, P‐gp, GSTP1, and Topo‐II Proteins in Non‐Small Cell Lung Cancer

Metastasis and multidrug resistance (MDR) are the main reasons for the poor prognosis of non‐small cell lung cancer (NSCLC) patients. The use of biomarkers may contribute to a more accurate prediction of tumor metastasis, a better response to chemotherapy, and better patient survival. Gelsolin‐like actin‐capping protein (CapG) and gelsolin have been identified as playing important roles in tumor invasion and metastasis. Permeability glycoprotein (P‐gp), glutathione S‐transferase pi (GSTP1), and topoisomerase‐II (Topo‐II) are proteins that are closely related to MDR. In this study, we assessed the prognostic significance of CapG and gelsolin (both markers of tumor motility), and of P‐gp, GSTP1, and Topo‐II (markers of MDR) in NSCLC patients. One hundred and twenty‐one patients with pathologically confirmed, resectable NSCLC were included in the study. The expression levels of the five kinds of proteins mentioned above were determined by immunohistochemistry (IHC). The correlation between the clinical characteristics and IHC findings were analyzed. Expression of CapG, gelsolin, and P‐gp was found to be associated with an increased risk of death (Hazard Ratio (HR) = 2.799, 95% Confidence Interval (CI) = 1.2705–6.169, P = 0.011; HR = 3.968, 95% CI = 1.811–8.693, P = 0.001; HR = 3.251, 95% CI = 1.456–7.260, P = 0.004, respectively), whereas expression of GSTP1 and Topo‐II was not. These results suggest that higher tumor motility and MDR may be important in NSCLC prognosis. Anat Rec, 2012.

Differential Expression of miR-125a-5p and let-7e Predicts the Progression and Prognosis of Non-Small Cell Lung Cancer

Aberrant expression of various microRNAs (miRNA) has shown diagnostic and prognostic significance in non-small cell lung cancer (NSCLC). qRT-PCR analysis confirmed altered expression of miR-125a-5p, let-7e, miR-30a, miR-30e and miR-30e-3p in 70 paired tissue and serum samples from NSCLC patients. The reduced expression of miR-125a-5p, let-7e and miR-30e was strongly associated with NSCLC dedifferentiation. The lost expression of miR-125a-5p and let-7e was associated with shorter overall survival and let-7e was an independent prognostic factor for NSCLC patients. These five miRNA expressions should be further evaluated as biomarkers for the early detection and prognosis of NSCLC patients.

Downregulation of miR-21 increases cisplatin sensitivity of non–small-cell lung cancer

Recent studies have shown that plasma miR-21 is a biomarker of chemotherapeutic response in lung cancer, but the influence of miR-21 on the sensitivity of non-small-cell lung cancer (NSCLC) to cisplatin (DDP) has not been confirmed. The aim of this study was to evaluate the role of miR-21 in NSCLC sensitivity to DDP in vitro and in vivo. Real-time quantitative PCR was used to detect miR-21 expression in lung cancer cell lines. Synthesized locked nucleic acid (LNA) anti-miR-21 was transiently transfected into A549 cells and pre-miR-21 was transfected into SK-MES-1 cells. We also investigated the effects of miR-21 downregulation and upregulation on growth and colony formation in DDP-treated cells. Finally, the effect of miR-21 downregulation on in vivo sensitivity of A549 cells to DDP was determined in BALB/c nude mice. miR-21 expression was significantly higher in A549 than in other lung cancer cell lines. LNA-based knockdown of miR-21 significantly inhibited growth and induced death in A549 cells, possibly via apoptotic signaling. Pre-miR-21 significantly promoted growth and inhibited death in SK-MES-1 cells. Moreover, ectopic suppression of miR-21 sensitized A549 cells to DDP in vivo. Our findings demonstrate that miR-21 suppression enhances the sensitivity of lung cancer cells to DDP in vitro and in vivo.

Hcmv-miR-UL112 Attenuates NK cell Activity by Inhibition Type I Interferon Secretion

Viral microRNAs (miRNAs) can regulate the host innate immune response. In particular, the human cytomegalovirus (HCMV) miRNA hcmv-miR-UL112 evades the host immune system by downregulating host immune gene and immediate-early viral gene expression. Natural killer (NK) cells are important innate immune cells with potent cytotoxicity, and are activated by type I interferons (IFNs) upon infection. It remains unclear how HCMV persists in the host despite the strongly antagonistic host immune system. A lentiviral vector was used to stably express hcmv-miR-UL112 in peripheral blood mononuclear cells (PBMCs). Hcmv-miR-UL112 expression levels were detected by TaqMan miRNA assay. The effects of hcmv-miR-UL112 on NK cell cytotoxicity were assessed with CD107a mobilization assay and CytoTox 96 non-radioactive cytotoxicity assay. Enzyme-linked immunosorbent assays (ELISA) were performed to detect type I IFNs levels in culture supernatants. To confirm the role of type I IFN in hcmv-miR-UL112-mediated NK cell cytotoxicity, PBMCs were incubated with antagonizing antibodies against IFN receptor (IFNAR) before lenti-hcmv-miR-UL112 treatment and recombinant type I IFN was added back into miR-transduced PBMC. Ectopically expressed hcmv-miR-UL112 functionally attenuated NK cell-mediated cytotoxicity, associated with decreased type I IFN expression. Hcmv-miR-UL112-transfected cells did not reduce the CD107-expression further than the IFNAR neutralizing mAbs-treatment alone, and adding back of recombinant type I IFN restored CD107a expression from the miR-transduced PBMC. Taken together, our results suggest that hcmv-miR-UL112 subverts innate immunity by downregulating type I IFN signaling to inhibit NK cell cytotoxicity. These results provide a new miRNA-based immunoevasion mechanism that may be exploited by HCMV.

Tim-3 Expression by Peripheral Natural Killer Cells and Natural Killer T Cells Increases in Patients with Lung Cancer - Reduction after Surgical Resection

BACKGROUND The purpose of this study was to investigate Tim-3 expression on peripheral CD3-CD56+ natural killer (NK) cells and CD3+CD56+ natural killer T (NKT) cells in lung cancer patients. MATERIALS AND METHODS We analyzed Tim-3+CD3-CD56+ cells, Tim-3+CD3-CD56dim cells, Tim-3+CD3-CD56bright cells, and Tim- 3+CD3+CD56+ cells in fresh peripheral blood from 79 lung cancer cases preoperatively and 53 healthy controls by flow cytometry. Postoperative blood samples were also analyzed from 21 members of the lung cancer patient cohort. RESULTS It was showed that expression of Tim-3 was significantly increased on CD3-CD56+ cells, CD3- CD56dim cells and CD3+CD56+ cells in lung cancer patients as compared to healthy controls (p=0.03, p=0.03 and p=0.04, respectively). When analyzing Tim-3 expression with cancer progression, results revealed more elevated Tim-3 expression in CD3-CD56+ cells, CD3-CD56dim cells and CD3+CD56+ cells in cases with advanced stages (III/IV) than those with stage I and II (p=0.02, p=0.04 and p=0.01, respectively). In addition, Tim-3 expression was significantly reduced on after surgical resection of the primary tumor (p<0.01). CONCLUSIONS Tim-3 expression in natural killer cells from fresh peripheral blood may provide a useful indicator of disease progression of lung cancer. Furthermore, it was indicated that Tim-3 might be as a therapeutic target.

Increased expression of T cell immunoglobulin and mucin domain 4 is positively associated with the disease severity of patients with ankylosing spondylitis.

Monocytes and associated cytokines have been shown to be involved in the pathogenesis of ankylosing spondylitis (AS). T cell immunoglobulin and mucin domain 4 (Tim-4) was identified on monocytes/macrophages and dentritic cells (DCs) and plays important roles in regulating the activities of macrophages and DCs. The current study investigated the association between Tim-4 expression and AS. Our results showed that Tim-4 expression on monocytes and Tim-4 level in plasma were highly increased in AS patients than in controls. Furthermore, TNF-α production and bath ankylosing spondylitis disease activity index (BASDAI) have positive relationships with Tim-4 expression in AS patients. High expression of Tim-4 was thought to contribute to the pathogenesis and an underlying mechanism of AS.