miR-125a-3p targets MTA1 to suppress NSCLC cell proliferation, migration, and invasion.

Metastasis-associated gene 1 (MTA1) is associated with cell growth, metastasis, and survival in non-small-cell lung cancer (NSCLC). Several previous reports have demonstrated that microRNAs affect gene expression through interaction between their seed region and the 3'-untranslated region of the target mRNA, resulting in post-transcriptional regulation. The aim of this study was to identify miRNAs that suppress malignancy in NSCLC cells by targeting MTA1. Two human NSCLC cell lines were analyzed for the expression of MTA1 by quantitative RT-PCR and western blotting after transfection with MTA1 mimics. A luciferase reporter assay was established to test the direct connection between MTA1 and its upstream miRNAs. Cell proliferation was assessed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay, 5-ethynyl-2'-deoxyuridine analysis, and colony formation assay. Cell migration and invasive capacity were evaluated by wound-healing assay and transwell assay. The miRNA/MTA1 axis was also probed by quantitative RT-PCR and western blotting in samples from eight NSCLC patients. Among the candidate miRNAs, miR-125a-3p was shown to post-transcriptionally regulate MTA1 in NSCLC cells. These data were reinforced by the luciferase reporter assay, in addition to the demonstration that MTA1 is inversely correlated with miR-125a-3p in NSCLC tissues. Furthermore, miR-125a-3p was found to inhibit NSCLC cell proliferation, migration, and invasion, through the same mechanisms of down-regulated MTA1. Our report demonstrates that miR-125a-3p inhibits the proliferation, migration, and invasion of NSCLC cells through down-regulation of MTA1, indicating the role of the miR-125a-3p/MTA1 axis in NSCLC, and may provide novel insight into the molecular mechanisms underpinning the disease and potential therapeutic targets.

Immune checkpoint inhibitors' combination therapy as first-line treatment in advanced esophageal squamous cell carcinoma: a meta-analysis.

PURPOSE: The benefit of immune checkpoint inhibitors' (ICIs) combination therapy in patients with advanced esophageal squamous cell carcinoma (ESCC) remained unclear. We performed a meta-analysis to explore the efficacy and safety of ICIs' combination therapy versus chemotherapy alone as first-line treatment in advanced ESCC. METHODS: A systematic review of randomized controlled trials (RCTs) of ICIs' combination therapy as first-line treatment in advanced ESCC was conducted via searching PubMed, Embase, and Cochrane database. The data for efficacy and safety of ICIs' combination therapy were subject to meta-analysis. Subgroup analysis was performed in patients with different PD-L1 expression status. RESULTS: A total of 5 RCTs and 3163 patients were included. Overall, the hazard ratio (HR) for overall survival (OS) benefit with ICIs' combination therapy was 0.68 (95% CI 0.62-0.75) compared with chemotherapy alone. The HR for progression-free survival (PFS) benefit and the odds ratio (OR) for overall response rate (ORR) increase were 0.62 (95% CI 0.56-0.68) and 2.01 (95% CI 1.70-2.38), respectively. The OS and PFS benefits with ICIs' combination therapy over chemotherapy alone were also observed in the subgroup of PD-L1 positive expression, but not in the subgroup of PD-L1 negative expression. The incidence of grade 3 or higher treatment-related adverse events was 60.4% with ICIs' combination therapy and 56.3% with chemotherapy alone (OR, 1.19; 95% CI 0.90-1.57). CONCLUSION: ICIs' combination therapy showed superior OS, PFS, and ORR over chemotherapy alone with a manageable safety profile. These results suggested that ICIs' combination therapy can be considered as a new first-line treatment for advanced ESCC.

An unresectable and metastatic intrahepatic cholangiocarcinoma with EML4-ALK rearrangement achieving partial response after first-line treatment with ensartinib: a case report.

Systemic chemotherapies are the primary treatment options for patients with unresectable and metastatic intrahepatic cholangiocarcinoma (ICC), but the effectiveness of current systemic therapies is limited. The development of targeted-therapy has changed the treatment landscape of ICC, and comprehensive genome sequencing of advanced cholangiocarcinoma patients could be beneficial to identify potential targets to guide individualized treatment. Herein, we reported an unresectable and metastatic ICC patient who detected EML4-ALK rearrangement in peripheral blood, which was later confirmed on tissue-based testing, and achieved partial response (PR) after first-line treatment with ensartinib. This case suggests that the liquid biopsy is of clinical value for unresectable or metastatic ICC, and the discovery of rare molecular targets provides new therapeutically approaches for advanced ICC patients.

Circulating miRNAs in Serum as Biomarkers for Early Diagnosis of Non-small Cell Lung Cancer.

BACKGROUND: Non-small cell lung cancer (NSCLC) accounts for about 85% of lung cancers. This study aimed to discover the potential miRNA biomarkers for early detection of NSCLC. METHODS: Total circulating miRNAs were extracted from six patients and six volunteers and run on the miRNA chip. The differentially expressed miRNAs acquired by data mining were intersected with chip results, and qRT-PCR were carried out. Then the differentially miRNAs were validated by using a validation cohort (120 participants). ROC curves were established to evaluate the diagnostic efficacy of the differentially circulating miRNAs. The target genes of the differential miRNAs were identified using the miRTarBase database, and follow-up GO and KEGG enrichment analysis were conducted. RESULTS: We identified 577 miRNA which screened according to the criteria (fold change > 2 and p value < 0.05). Among them, seven circulating miRNAs passed additional filtering based on data mining. These miRNAs were further validated in the training and validation cohort. miR-492, miR-590-3p, and miR-631 were differentially expressed in the patients' serum, and the area under the ROC curve (AUC) values of these miRNAs were 0.789, 0.792, and 0.711, respectively. When using them as a combination to discriminate healthy volunteers from patients, the AUC reached 0.828 (95% CI, 0.750-0.905, p = 0.000) with a sensitivity of 86.7% and specificity of 71.7%. The follow-up enrichment analysis showed that target genes of three miRNA were associated with tumorigenesis and progression, such as cell cycle and P53 signaling pathway. CONCLUSIONS: The combination of miR-492, miR-590-3p, and miR-631 can be utilized to distinguish healthy individuals and early-stage NSCLC patients. IMPACT: The combination of miR-492, miR-590-3p, and miR-631 might be a promising serum biomarker in patients for the early diagnosis of NSCLC.

Prognostic factors of oligometastatic non-small cell lung cancer: a meta-analysis.

BACKGROUND: The prognostic factors of oligometastatic non-small cell lung cancer (NSCLC) are uncertain. We performed a meta-analysis to assess the prognostic factors of oligometastatic NSCLC patients who are most likely to achieve long-term survival. METHODS: We searched PubMed, EMBASE, the Cochrane to identify eligible articles and performed the meta-analysis of all randomized controlled trials (RCTs) and retrospective comparative studies revealing the prognostic factors of oligometastatic NSCLC. The primary endpoint of interest was overall survival (OS). RESULTS: We analyzed data from twenty-four eligible studies, including data from 1,935 patients with oligometastatic NSCLC. In the univariate analysis, we found no significant difference in OS of prognostic factors including age [hazard ratios (HRs) 1.02, 95% CI: 0.80-1.31, P=0.86], smoking status (HR 1.08, 95% CI: 0.80-1.46, P=0.62), type of metastases (HR 1.61, 95% CI: 0.86-3.03, P=0.14), but significantly positive prognoses containing female (HR 1.21, 95% CI: 1.02-1.45, P=0.03), (y)pN0 stage (HR 1.82, 95% CI: 1.40-2.36, P<0.00001), adenocarcinoma (HR 1.44, 95% CI: 1.10-1.88, P=0.008). In the multivariate analysis, patients with (y)pN0 stage had an obvious survival benefit compared with (y)pN1 (HR 1.63, 95% CI: 1.27-2.10, P=0.001), but no significant survival in contrast with (y)pN2 (HR 2.01, 95% CI: 0.80-5.03, P=0.14). In subgroup analyses, neither thoracic stage (HR 2.06, 95% CI: 1.52-2.78, P=0.55), (y)pT-stage of primary lung cancer (HR 1.38, 95% CI: 0.86-2.21, P=0.14) nor tumorous histology (HR 2.99, 95% CI: 2.10-4.28, P=0.91) and oligometastatic number (HR 1.25, 95% CI: 0.97-1.62, P=0.98) were significantly different in OS. However, patients with aggressive thoracic treatment (ATT) had improved survival (HR 0.56, 95% CI: 0.37-0.83, P=0.001), and notably, different strategies of ATT received by oligometastatic NSCLC patients might significantly influence survival (HR 0.54, 95% CI: 0.36-0.82, P<0.00001). CONCLUSIONS: Overall, factors including age, smoking status, type of metastasis were not associated with long-term survival of oligometastatic NSCLC patients. However, our finding suggests that aggressive therapies in the primary lung cancer, as well as female, (y)pT-stage, absence of nodal diseases, adenocarcinoma histology have been clarified as positive prognosis. Further studies of prospective study for these patients are warranted.

HOXA10­AS: A novel oncogenic long non­coding RNA in glioma.

Glioma is the most common primary malignant tumor of the central nervous system. Emerging evidence has demonstrated that long non­coding RNAs (lncRNAs) serve a major role of regulation in various types of human cancer, including glioma. However, the biological roles of thousands of lncRNAs remain unknown and require further identification. The present study investigated the functional role of lncRNA­HOXA10­AS in glioma. The present study examined the expression patterns of HOXA10­AS in glioma and normal brain tissues, as well as glioma cell lines and normal human astrocytes (HA) via reverse transcription­quantitative polymerase chain reaction. HOXA10­AS knockdown cells were generated using lentiviral short hairpin RNA against HOXA10­AS in A172 and U251 glioma cells. Cell growth was assessed by MTT assay, and a flow cytometer was used to investigate cell proliferation, cell cycle distribution and cell apoptosis. Western blot analysis was performed to analyze the expression levels of apoptosis­related proteins. HOXA10­AS was significantly upregulated in glioma tissues and cell lines, and increased HOXA10­AS expression levels were associated with higher grades of glioma. Knockdown of HOXA10­AS inhibited glioma cell proliferation and increased cell apoptosis rates compared with the control cells. HOXA10­AS markedly regulated the expression of the homeobox A10 (HOXA10) gene. Similarly, HOXA10 expression was increased with higher grades of glioma, and silencing of HOXA10 by small interfering RNA suppressed glioma cell proliferation and induced cell apoptosis. The results of the present study demonstrated that HOXA10­AS promoted cell growth and survival through activation of HOXA10 gene expression in glioma, which may potentially act as a novel biomarker and therapeutic target for clinical assay development.

Effect of postoperative radiotherapy on outcome in resectable stage IIIA-N2 non-small-cell lung cancer: an updated meta-analysis.

BACKGROUND: In a previous meta-analysis-based modeling study, it was hypothesized that modern postoperative radiotherapy (PORT) may improve both local recurrence and overall survival (OS) in stage IIIA-N2 non-small-cell lung cancer (NSCLC). There were only four single-arm trials with a total of 357 patients. As other trials have provided new and controversial data, we performed this updated meta-analysis to test the hypothesis. PATIENTS AND METHODS: Systematic reviews in Medline, Cochrane, and Science Direct up to December 2015 identified publications exploring the efficacy of PORT in resectable stage IIIA-N2 NSCLC. RESULTS: Overall, 16 trials comprising 3278 patients were included. There was a significant benefit in favor of PORT regarding OS [hazard ratio (HR): 0.73, 95% confidence interval (CI): 0.58-0.92, P=0.008; absolute benefit at 5 years=8%], disease-free survival (HR: 0.70, 95% CI: 0.60-0.83, P<0.0001), and locoregional recurrence-free survival (HR: 0.37, 95% CI: 0.24-0.58, P<0.0001). Restriction of the analysis to trials with induction and/or adjuvant chemotherapy led to similar results. PORT significantly decreased the risk of local recurrence (risk ratio: 0.64, 95% CI: 0.50-0.82, P=0.0006) and DM (risk ratio: 0.74, 95% CI: 0.62-0.88, P=0.0005), and the absolute risk differences were 13 and 14%, respectively. CONCLUSION: The addition of PORT, with or without chemotherapy, significantly improves local control and survival in patients with resectable stage IIIA-N2 NSCLC.

Should aggressive thoracic therapy be performed in patients with synchronous oligometastatic non-small cell lung cancer? A meta-analysis.

BACKGROUND: We performed a meta-analysis to compare overall survival (OS) outcomes in patients with synchronous oligometastatic non-small cell lung cancer (NSCLC) who underwent aggressive thoracic therapy (ATT) with those who did not. METHODS: A systematic review of controlled trials of ATT on survival in synchronous oligometastatic NSCLC was conducted. Hazard ratio (HR) for the main endpoint OS was pooled using a fixed-effects model. Subgroup analysis was performed in patients with single organ metastases, or with different numbers of brain metastases, or with different stages of thoracic disease. Pooled survival curves of OS were constructed. RESULTS: Seven eligible retrospective observational cohort studies were identified including 668 synchronous oligometastatic NSCLC patients, of whom 227 (34.0%) received ATT. For patients with synchronous oligometastatic NSCLC, ATT was associated with a significant improvement of OS (HR, 0.48; 95% CI, 0.39-0.60; P<0.00001). In subgroup analysis, the association with OS was similar or even strengthened, with a HR of 0.42 (95% CI, 0.31-0.56) in single organ metastases group, 0.49 (95% CI, 0.31-0.75) in solitary brain metastasis group, and 0.38 (95% CI, 0.20-0.73) in thoracic stage I-II group, respectively. The pooled cumulative survival rates for patients received ATT were 74.9% at 1 year, 52.1% at 2 years, 23.0% at 3 years, and 12.6% at 4 years. The corresponding pooled survival for patients who did not receive ATT were 32.3%, 13.7%, 3.7%, and 2.0%, respectively. CONCLUSIONS: Survival benefit from ATT is common in synchronous oligometastatic patients. Selected patients with synchronous oligometastatic NSCLC could also achieve long-term survival with ATT.

Optimization of radiotherapy for neck carcinoma metastasis from unknown primary sites: a meta-analysis.

This meta-analysis was designed to evaluate radiotherapy (RT) options preferable for neck cancer metastases from unknown primary sites (NCUP). Relevant articles published up through September 2015 were selected from EMBASE, Cochrane, PubMed and Web of Science. Thirty-three articles were identified, and relative risks (RRs) and 95% CIs for all pre-specified endpoints were calculated. Surgery plus RT showed an advantage for 5-year overall survival (OS) (RR 0.66, 95% CI 0.52-0.83, p = 0.0004) and neck recurrence (NR) (RR = 0.74, 95% CI 0.59-0.92, p = 0.008) compared to RT alone. The RRs for NR, primary tumor emergence (PTE), and 5-year disease free survival (DFS) for bilateral neck compared to ipsilateral neck irradiation were 0.61 (95% CI 0.41-0.91, p = 0.01), 0.44(95% CI 0.26-0.77, p = 0.004), and 0.81 (95% CI 0.64-1.03, p = 0.09), respectively. Irradiation of the neck plus potential primary tumor sites (PPTS) showed a benefit for 5-year DFS (RR 0.75, 95% CI 0.61-0.92, p = 0.005), NR (RR = 0.72, 95% CI 0.56-0.92, p = 0.009), and PTE (RR = 0.23, 95% CI 0.12-0.45, p < 0.0001) compared to neck-only irradiation. Adverse events occurred more frequently with bilateral neck plus PPTS irradiation. For NCUP, surgery plus RT of the bilateral neck and PPTS was associated with greater improvement of clinical outcomes.

Proto-oncogenic miR-744 is upregulated by transcription factor c-Jun via a promoter activation mechanism.

Upregulation of miR-744 is associated with poor prognosis in many types of cancer patients, but it is still unclear how miR-744 becomes elevated in these tumors. In this study, we found that ectopic c-Jun elevated miR-744 expression, whereas c-Jun attenuation reduced miR-744 expression. Chromatin immunoprecipitation assay confirmed the direct binding of c-Jun to the promoter of miR-744. The binding site of -343 to -349 bp within the most potential promoter like sequence of miR-744 was further validated by luciferase reporter gene assays. C-Jun-induced miR-744 upregulation could significantly promote migration and invasion of nasopharyngeal carcinoma cells and non-small cell lung cancer (NSCLC) cells, hence ectopic c-Jun was sufficient to rescue the migratory and invasive ability of these cells when miR-744 was knockdown. Additionally, a positive correlation between the expression levels of miR-744 and c-Jun was revealed in NSCLC samples with high (top 10%) level of miR-744 expression from the TCGA dataset. Taken together, our results demonstrated for the first time the regulatory mechanism of miR-744 transcription by c-Jun, providing a potential mechanism underlying the upregulation of miR-744 in cancers.

MiR-744 functions as a proto-oncogene in nasopharyngeal carcinoma progression and metastasis via transcriptional control of ARHGAP5.

Nasopharyngeal carcinoma (NPC) is a highly invasive and metastasis-prone epithelial cancer. The paucity of effective treatment strategies for recurrent and metastatic NPC is the major cause for stagnating survival rate of NPC. Therefore, it's urgent to understand the molecular mechanisms underlying NPC progression and identify novel avenues for targeted therapy. It has emerged recently that microRNAs are potential pro-tumorigenic or tumor-suppressive factors that participate in oncogenesis. In this study, we found that miR-744 expression was upregulated in NPC specimens compared to nasopharyngeal epithelium (NPE) tissue, and miR- 744 upregulation was significantly associated with TNM stage, tumorigenesis and metastasis. Functional studies revealed that miR-744 acts as a novel tumor promotor in NPC. Moreover, we determined that miR-744 targets ARHGAP5 (Rho GTPase activating protein 5), a protumorigenic gene, by directly interacting with its promoter and thereby regulating its expression at transcriptional level. Reintroduction of ARHGAP5 resembled the effects of miR-744 and silencing of ARHGAP5 clearly abrogated miR-744-induced enhancement of cell migration and invasion. High level of ARHGAP5 was positively correlated with that of miR-744 and with advanced stages of NPC, as well as with lymph node metastasis. Taken together, these data reveal for the first time that miR-744 exerts its proto-oncogenic function by directly targeting ARHGAP5 promoter. This newly identified miR-744/ARHGAP5 pathway provides further insight into the progression and metastasis of NPC and indicates potential novel therapeutic targets for NPC.

KRAS Mutation as a Biomarker for Survival in Patients with Non-Small Cell Lung Cancer, A Meta-Analysis of 12 Randomized Trials.

BACKGROUND: Because there is no clear consensus for the prognostic implication of KRAS mutations in patients with non-small cell lung cancer (NSCLC), we conducted a meta-analysis based on 12 randomized trials to draw a more accurate conclusion. MATERIALS AND METHODS: A systematic computer search of articles from inception to May 1, 2014 using the PubMed, EMBASE, and Cochrane databases was conducted. The enrollment of articles and extraction of data were independently performed by two authors. RESULTS: Our analysis was based on the endpoints overall survival (OS) and progression-free survival (PFS). Nine records (All for OS, 7 for PFS) comprising 12 randomized trials were identified with 3701 patients who underwent a test for KRAS mutations. In the analysis of the pooled hazard ratios (HRs) for OS (HR: 1.39; 95% confidence interval [CI] 1.23-1.56) and PFS (HR: 1.33; 95% CI 1.17-1.51), we found that KRAS mutations are related to poor survival benefit for NSCLC. According to a subgroup analysis stratified by disease stage and line of therapy, the combined HRs for OS and PFS coincided with the finding that the presence of a KRAS mutation is a dismal prognostic factor. However, the prognostic role of KRAS mutations are not statistically significant in a subgroup analysis of patients treated with chemotherapy in combination with cetuximab based on the endpoints OS (P=0.141) and PFS (P=0.643). CONCLUSIONS: Our results indicate that KRAS mutations are associated with inferior survival benefits for NSCLC but not for those treated with chemotherapies integrating cetuximab.