Bingyin Shi

Highly frequent PIK3CA amplification is associated with poor prognosis in gastric cancer

BackgroundThe phosphoinositide 3-kinase (PI3K)/Akt pathway plays a fundamental role in cell proliferation and survival in human tumorigenesis, including gastric cancer. PIK3CA mutations and amplification are two major causes of overactivation of this pathway in human cancers. However, until this work, there was no sound investigation on the association of PIK3CA mutations and amplification with clinical outcome in gastric cancer, particularly the latter.MethodsUsing direct sequencing and real-time quantitative PCR, we examined PIK3CA mutations and amplification, and their association with clinicopathological characteristics and clinical outcome of gastric cancer patients.ResultsPIK3CA mutations and amplification were found in 8/113 (7.1%) and 88/131 (67%) gastric cancer patients, respectively. PIK3CA amplification was closely associated with increased phosphorylated Akt (p-Akt) level. No relationship was found between PIK3CA mutations and clinicopathological characteristics and clinical outcome in gastric cancer. PIK3CA amplification was significantly positively associated with cancer-related death. Importantly, Kaplan-Meier survival curves revealed that the patients with PIK3CA amplification had significantly shorter survival times than the patients without PIK3CA amplification.ConclusionsOur data showed that PIK3CA mutations were not common, but its amplification was very common in gastric cancer and may be a major mechanism in activating the PI3K/Akt pathway in gastric cancer. Importantly, Kaplan-Meier survival curves revealed that PIK3CA amplification was significantly positively associated with poor survival of gastric cancer patients. Collectively, the PI3K/Akt signaling pathway may be an effective therapeutic target in gastric cancer.

Highly frequent promoter methylation and PIK3CA amplification in non-small cell lung cancer (NSCLC)

BackgroundLung cancer is the leading cause of cancer-related death worldwide. Genetic and epigenetic alterations have been identified frequently in lung cancer, such as promoter methylation, gene mutations and genomic amplification. However, the interaction between genetic and epigenetic events and their significance in lung tumorigenesis remains poorly understood.MethodsWe determined the promoter methylation of 6 genes and PIK3CA amplification using quantitative methylation-specific PCR (Q-MSP) and real-time quantitative PCR, respectively, and explore the association of promoter methylation with PIK3CA amplification in a large cohort of clinically well-characterized non-small cell lung cancer (NSCLC).ResultsHighly frequent promoter methylation was observed in NSCLC. With 100% diagnostic specificity, excellent sensitivity, ranging from 45.8 to 84.1%, was found for each of the 6 genes. The promoter methylation was associated with histologic type. Methylation of CALCA, CDH1, DAPK1, and EVX2 was more common in squamous cell carcinomas (SCC) compared to adenocarcinomas (ADC). Conversely, there was a trend toward a higher frequency of RASSF1A methylation in ADC than SCC. In addition, PIK3CA amplification was frequently found in NSCLC, and was associated with certain clinicopathologic features, such as smoking history, histologic type and pleural indentation. Importantly, aberrant promoter methylation of certain genes was significantly associated with PIK3CA amplification.ConclusionsOur data showed highly frequent promoter methylation and PIK3CA amplification in Chinese NSCLC population, and first demonstrated the associations of gene methylation with PIK3CA amplification, suggesting that these epigenetic events may be a consequence of overactivation of PI3K/Akt pathway.

Low Population Selenium Status Is Associated With Increased Prevalence of Thyroid Disease

CONTEXT Epidemiological studies have supported the premise that an adequate selenium intake is essential for thyroid gland function. OBJECTIVE The objective was to investigate whether the prevalence of thyroid disease differed in two areas that were similar, except for very different soil/crop selenium concentrations. DESIGN Cross-sectional observational study. SETTING The setting was two counties of Shaanxi Province, China, here defined as adequate- and low-selenium. PARTICIPANTS A total of 6152 participants were selected by stratified cluster-sampling. MAIN OUTCOME MEASURES Participants completed demographic and dietary questionnaires and underwent physical and thyroid ultrasound examinations. Serum samples were analyzed for thyroid function parameters and selenium concentration. Serum selenium was compared between different demographic, dietary, and lifestyle categories in the two counties. The relationship between selenium status, dietary factors, and pathological thyroid conditions was explored by logistic regression. RESULTS Complete data sets were available from 3038 adequate-selenium participants and 3114 low-selenium participants in whom median (interquartile range) selenium concentrations differed almost 2-fold (103.6 [79.7, 135.9] vs 57.4 [39.4, 82.1] μg/L; P = .001). The prevalence of pathological thyroid conditions (hypothyroidism, subclinical hypothyroidism, autoimmune thyroiditis, and enlarged thyroid) was significantly lower in the adequate-selenium county than in the low-selenium county (18.0 vs 30.5%; P < .001). Higher serum selenium was associated with lower odds ratio (95% confidence interval) of autoimmune thyroiditis (0.47; 0.35, 0.65), subclinical hypothyroidism (0.68; 0.58, 0.93), hypothyroidism (0.75; 0.63, 0.90), and enlarged thyroid (0.75; 0.59, 0.97). CONCLUSIONS Low selenium status is associated with increased risk of thyroid disease. Increased selenium intake may reduce the risk in areas of low selenium intake that exist not only in China but also in many other parts of the world.

Quantitative assessment of gene methylation and their impact on clinical outcome in gastric cancer.

BACKGROUND Promoter methylation is an alternative mechanism of gene silencing in human tumorigenesis. Although a number of methylated genes have been found in gastric cancer, useful methylation markers for early diagnosis and prognostic evaluation of this cancer remain largely unknown. METHODS Using quantitative methylation-specific PCR (Q-MSP), we examined promoter methylation of 6 genes, including CALCA, DAPK1, RARbeta, RASSF1A, TIMP3, and PAX6, and explored their association with clinical outcome in gastric cancer. RESULTS We found that most of the genes investigated in the present study had significantly higher methylation level in tumor tissues than normal gastric tissues, including CALCA, RARbeta, RASSF1A, TIMP3, and PAX6. With more focus on specificity compared to sensitivity, all genes were hypermethylated in gastric cancer, ranging from 12.8% to 36.9%. Methylation of TIMP3 and PAX6 was strongly associated with differentiation and lymph node metastasis, respectively. Importantly, most of gene methylation, except for DAPK1, was closely associated with poor survival in gastric cancer. CONCLUSION We found that a panel of genes was specifically methylated in gastric cancer, and demonstrated the effect of promoter methylation of some genes on clinical outcome in gastric cancer, indicating these methylated genes may be useful biomarkers for prognostic evaluation in this cancer.

Frequent Gene Amplification Predicts Poor Prognosis in Gastric Cancer

Gastric cancer is one of the most common malignancies worldwide. However, genetic alterations leading to this disease are largely unknown. Gene amplification is one of the most frequent genetic alterations, which is believed to play a major role in the development and progression of gastric cancer. In the present study, we identified three frequently amplified genes from 30 candidate genes using real-time quantitative PCR method, including ERBB4, C-MET and CD44, and further explored their association with clinicopathological characteristics and poor survival in a cohort of gastric cancers. Our data showed amplification of these genes was significantly associated with certain clinicopathological characteristics, particularly tumor differentiation and cancer-related death. More importantly, amplification of these genes was significantly related to worse survival, suggesting that these amplified genes may be significant predictors of poor prognosis and potential therapeutic targets in gastric cancer. Targeting these genes may thus provide new possibilities in the treatment of gastric cancer.

Gut microbiota as a potential target of metabolic syndrome: the role of probiotics and prebiotics

Metabolic syndrome (MS) comprises central obesity, increased plasma glucose levels, hyperlipidemia and hypertension, and its incidence is increasing due to changes in lifestyle and dietary structure in recent years. MS has been proven to be associated with an increased incidence of cardiovascular diseases and type 2 diabetes mellitus, leading to morbidity and mortality. In this manuscript, we review recent studies concerning the role of the gut microbiota in MS modulation. Manipulation of the gut microbiota through the administration of prebiotics or probiotics may assist in weight loss and reduce plasma glucose and serum lipid levels, decreasing the incidence of cardiovascular diseases and type 2 diabetes mellitus. To the best of our knowledge, short-chain fatty acids (SCFAs), bile salt hydrolase (BSH), metabolic endotoxemia and the endocannabinoid (eCB) system are essential in regulating the initiation and progression of MS through the normalization of adipogenesis and the regulation of insulin secretion, fat accumulation, energy homeostasis, and plasma cholesterol levels. Therefore, the gut microbiota may serve as a potential therapeutic target for MS. However, further studies are needed to enhance our understanding of manipulating the gut microbiota and the role of the gut microbiota in MS prevention and treatment.

Metallothionein 1G functions as a tumor suppressor in thyroid cancer through modulating the PI3K/Akt signaling pathway

BackgroundMT1G inactivation mediated by promoter methylation has been reported in thyroid cancer. However, the role of MT1G in thyroid carcinogenesis remains unclear. The aim of this study is to examine the biological functions and related molecular mechanisms of MT1G in thyroid cancer.MethodsMethylation-specific PCR (MSP) was performed to analyze promoter methylation of MT1G and its relationship with clinicopathological characteristics of papillary thyroid cancer (PTC) patients. Conventional and real-time quantitative RT-PCR assays were used to evaluate mRNA expression. The functions of ectopic MT1G expression were determined by cell proliferation and colony formation, cell cycle and apoptosis, as well as cell migration and invasion assays.ResultsMT1G expression was frequently silenced or down-regulated in thyroid cancer cell lines, and was also significantly decreased in primary thyroid cancer tissues compared with non-malignant thyroid tissues. Promoter methylation, along with histone modification, contributes to MT1G inactivation in thyroid tumorigenesis. Moreover, our data showed that MT1G hypermethylation was significantly positively associated with lymph node metastasis in PTC patients. Importantly, restoring MT1G expression in thyroid cancer cells dramatically suppressed cell growth and invasiveness, and induced cell cycle arrest and apoptosis through inhibiting phosphorylation of Akt and Rb.ConclusionsWe have for the first time revealed that MT1G appears to be functional tumor suppressor involved in thyroid carcinogenesis mainly through modulating the phosphatidylinositol-3-kinase (PI3K)/Akt pathway and partially through regulating the activity of Rb/E2F pathway in this study.

Sulforaphane inhibits thyroid cancer cell growth and invasiveness through the reactive oxygen species-dependent pathway

Sulforaphane (SFN), a natural compound derived from broccoli/broccoli sprouts, has been demonstrated to be used as an antitumor agent in different types of cancers. However, its antitumor effect in thyroid cancer remains largely unknown. The aim of the study was to investigate the therapeutic potential of SFN for thyroid cancer and explore the mechanisms underlying antitumor effects of SFN by in vitro and in vivo studies. Our data demonstrated that SFN significantly inhibited thyroid cancer cell proliferation in a dose- and time-dependent manner, induced G2/M phase cell cycle arrest and apoptosis, and inhibited thyroid cancer cell migration and invasion by suppressing epithelial-mesenchymal transition (EMT) process and expression of Slug, Twist, MMP-2 and -9. Mechanically, SFN inhibited thyroid cancer cell growth and invasiveness through repressing phosphorylation of Akt, enhancing p21 expression by the activation of Erk and p38 signaling cascades, and promoting mitochondrial-mediated apoptosis via reactive oxygen species (ROS)-dependent pathway. Growth of xenograft tumors derived from thyroid cancer cell line FTC133 in nude mice was also significantly inhibited by SFN. Importantly, we did not find significant effect of SFN on body weight and liver function of mice. Collectively, we for the first time demonstrate that SFN is a potentially effective antitumor agent for thyroid cancer.

TERT promoter mutations and long telomere length predict poor survival and radiotherapy resistance in gliomas

Increasing evidences have implicated somatic gain-of-function mutations at the telomerase reverse transcriptase (TERT) promoter as one of the major mechanisms that promote transcriptional activation of TERT and subsequently maintain telomere length in human cancers including glioma. To investigate the prognostic value of these mutations and telomere length, individually and their coexistence, in gliomas, we analyzed two somatic mutations C228T and C250T in the TERT promoter, relative telomere length (RTL), IDH1 mutation and MGMT methylation in 389 glioma patients, and explored their associations with patient characteristics and clinical outcomes. Our data showed that C228T and C250T mutations were found in 17.0% (66 of 389) and 11.8% (46 of 389) of gliomas, respectively, and these two mutations were mutually exclusive in this cancer. Moreover, they were significantly associated with WHO grade. We also found that the RTL was significant longer in gliomas than in meningiomas and normal brain tissues (Median, 0.89 vs. 0.44 and 0.50; P < 0.001), and demonstrated that the RTL was strongly correlated with tumor recurrence. Importantly, TERT promoter mutations or long RTL caused a significantly poorer survival than TERT wild-type or short RTL. Coexisting TERT promoter mutations and long RTL were more commonly associated with poor patient survival than they were individually. Notably, the patients with TERT promoter mutations particularly C228T or long RTL were resistant to radiotherapy. Collectively, TERT promoter mutations and long RTL are not only prognostic factors for poor clinical outcomes, but also the predictors of radiotherapy resistance in gliomas.

Low frequency of TERT promoter mutations in a large cohort of gallbladder and gastric cancers

Dear Editors, Telomerase is a ribonucleoprotein that consists of an RNA subunit and a reverse transcriptase catalytic subunit. The latter is encoded by telomerase reverse transcriptase (TERT) gene. Telomerase adds telomeric repeats to chromosome ends, thereby maintaining telomere length. It is frequently upregulated in human cancers, contributing to human tumorigenesis. Mutations in the TERT gene promoter have been reported in melanoma, follicular cellderived thyroid cancer, bladder cancer, glioblastoma and other cancers. Two of them, 1,295,228 C>T and 1,295,250 C>T (termed C228T and C250T hereafter, respectively), are particularly common. These two mutations have been demonstrated to confer the TERT promoter increased transcriptional activity. Importantly, these mutations are found to be absent in benign tumors and normal human subjects, further implicating their potentially critical roles in human tumorigenesis. However, until now, TERT promoter mutations have not been investigated in a large cohort of gallbladder and gastric cancers. Although a previous study has shown no mutations in TERT gene promoter in gallbladder cancer, the limited number (N5 10) yielded an inconclusive results. In this study, a total of 154 gallbladder cancer tissues and 268 gastric cancer tissues were randomly obtained from the First Affiliated Hospital of Xi’an Jiaotong University School of Medicine, with approval by the institutional review board of the Hospital. Clinicopathological data were obtained from the patients’ files or by interview with the patients or their relatives, and were presented in Supporting Information Table 1. Genomic DNA was isolated using standard procedures of protease K digestion, phenol-chloroform extraction and ethanol precipitation. A pyrosequencing assay was performed to examine these two TERT promoter mutations. The following primers were used to amplify a fragment of 162 bp, which contained the sites of C228T and C250T mutations: 50-GCG CTG CCT GAA ACT CGC-30 (sense, biotinylated at the 50-end) and 50-CGT CCT GCC CCT TCA CCT-30 (antisense). PCR was performed with an initial denaturation at 95 C for 3 min, followed by 40 cycles of 95 C denaturation for 30 sec, 59 C annealing for 30 sec and 68 C elongation for 1 min. Quality of PCR products was determined by gel electrophoresis. The primer for pyrosequencing (50-CCC GCC CCC TCC CGA-30) was designed immediately upstream of C250T so that these two mutations are analyzed in the same assay. Pyrosequencing assay was performed on a PyroMark Q24 system using PyroMark Gold Q24 reagent (Qiagen). Sanger sequencing was then performed to confirm the results of pyrosequencing using the same primer pair as for pyrosequencing assay. As shown in Figure 1 and Table 1, similar to the findings in a previous study, the frequency of TERT promoter mutations in esophageal squamous cell carcinoma (ESCC) was very low, we also found low frequency of TERT promoter mutations in primary gallbladder and gastric cancer tissues, and gastric cancer cell lines. C228T and C250T mutations were found in 6 and 8 of 154 (3.9% and 5.2%) gallbladder cancer tissues, respectively, and these two mutations were mutually exclusive in this cancer. Notably, only two C250T mutations were found in 268 gastric cancer tissues, and no C228T or C250T mutations were found in eight gastric cancer cell lines. Given extremely low frequency of TERT promoter mutations in gastric cancer, we only investigated the association of these two mutations with clinicopathological characteristics of gallbladder cancer patients, including gender, age, tumor size, invasion, tumor node metastasis (TNM) stage, lymph node metastasis and survival status. The data showed that TERT promoter mutations did not significantly correlate with any clinicopathological variables in gallbladder cancer (Supporting Information Table 2).