Efficient detection and post-surgical monitoring of colon cancer with a multi-marker DNA methylation liquid biopsy.

Multiplex assays, involving the simultaneous use of multiple circulating tumor DNA (ctDNA) markers, can improve the performance of liquid biopsies so that they are highly predictive of cancer recurrence. We have developed a single-tube methylation-specific quantitative PCR assay (mqMSP) that uses 10 different methylation markers and is capable of quantitative analysis of plasma samples with as little as 0.05% tumor DNA. In a cohort of 179 plasma samples from colorectal cancer (CRC) patients, adenoma patients, and healthy controls, the sensitivity and specificity of the mqMSP assay were 84.9% and 83.3%, respectively. In a head-to-head comparative study, the mqMSP assay also performed better for detecting early-stage (stage I and II) and premalignant polyps than a published SEPT9 assay. In an independent longitudinal cohort of 182 plasma samples (preoperative, postoperative, and follow-up) from 82 CRC patients, the mqMSP assay detected ctDNA in 73 (89.0%) of the preoperative plasma samples. Postoperative detection of ctDNA (within 2 wk of surgery) identified 11 of the 20 recurrence patients and was associated with poorer recurrence-free survival (hazard ratio, 4.20; P = 0.0005). With subsequent longitudinal monitoring, 14 patients (70%) had detectable ctDNA before recurrence, with a median lead time of 8.0 mo earlier than seen with radiologic imaging. The mqMSP assay is cost-effective and easily implementable for routine clinical monitoring of CRC recurrence, which can lead to better patient management after surgery.

Deubiquitinase inhibitor degrasyn suppresses metastasis by targeting USP5-WT1-E-cadherin signalling pathway in pancreatic ductal adenocarcinoma.

Wilm's tumour-1 (WT1) is overexpressed in pancreatic ductal adenocarcinoma (PDAC) and enhances metastasis. Deubiquitination stabilizes target proteins, and inhibiting deubiquitination facilitates the degradation of target proteins. However, whether inhibiting deubiquitination of WT1 facilitates its degradation and presents anti-cancer ability in PDAC is unknown. Here, we found that deubiquitinase inhibitor degrasyn rapidly induced the degradation of endogenous and exogenous WT1 through enhancing ubiquitination of WT1 followed by the up-regulation of E-cadherin. Knockdown of WT1 by short hairpin RNAs (shRNAs) inhibited metastasis and overexpression of WT1 partially prevented degrasyn-induced anti-metastasis activity, suggesting that degrasyn presents anti-metastasis activity partially through degrading WT1 protein. We further identified that USP5 deubiquitinated WT1 and stabilized its expression. The higher expressions of USP5 and WT1 are associated with tumour metastasis. More importantly, degrasyn inhibited the activity of USP5 and overexpression of USP5 partially prevented degrasyn-induced degradation of WT1 protein, suggesting that degrasyn degraded WT1 protein through inhibiting the activity of USP5. Finally, degrasyn reduced the tumorigenicity in a xenograft mouse model and reduced the metastasis in vivo. Our results indicate that degrasyn presents strong anti-cancer activity through USP5-WT1-E-cadherin signalling in PDAC. Therefore, degrasyn holds promise as cancer therapeutic agent in PDAC with high expressions of USP5 and WT1.

Inhibition of UBE2N-dependent CDK6 protein degradation by miR-934 promotes human bladder cancer cell growth.

Because bladder cancer (BC) is one of the most common malignant cancers of the urinary system, identification of BC cell growth-associated effectors is of great significance. Cyclin-dependent kinase (CDK)6 is a member of the CDK family of cell cycle-related proteins and plays an important role in cancer cell growth. This is borne out by the fact that a CDK6 inhibitor had been approved to treat several types of cancers. Nevertheless, underlying molecular mechanisms concerning how to regulate CDK6 expression in BC remains unclear. In the present study, it was observed that miR-934 was much higher in human BCs and human BC cell lines as well. The results also revealed that miR-934 inhibition dramatically decreased human BC cell monolayer growth in vitro and xenograft tumor growth in vivo; the outcomes were accompanied by CDK6 protein down-regulation and G0-G1 cell cycle arrest. Moreover, overexpression of CDK6 reversed the inhibition of BC cell growth induced by miR-934. Further studies showed that miR-934 binds to a 3'-UTR of ubiquitin-conjugating enzyme 2N (ube2n) mRNA, down-regulated UBE2N protein expression; this, in turn, attenuated CDK6 protein degradation and led to CDK6 protein accumulation as well as the promotion of BC tumor growth. Collectively, this study not only establishes a novel regulatory axis of miR-934/UBE2N of CDK6 but also provides data suggesting that miR-934 and UBE2N may be potentially promising targets for therapeutic strategies against BC.-Yan, H., Ren, S., Lin, Q., Yu, Y., Chen, C., Hua, X., Jin, H., Lu, Y., Zhang, H., Xie, Q., Huang, C., Huang, H. Inhibition of UBE2N-dependent CDK6 protein degradation by miR-934 promotes human bladder cancer cell growth.

miR-3687 Overexpression Promotes Bladder Cancer Cell Growth by Inhibiting the Negative Effect of FOXP1 on Cyclin E2 Transcription.

Cyclin E2, a member of the cyclin family, is a key cell cycle-related protein. This protein plays essential roles in cancer progression, and, as such, an inhibitor of cyclin E2 has been approved to treat several types of cancers. Even so, mechanisms underlying how to regulate cyclin E2 expression in cancer remain largely unknown. In the current study, miR-3687 was upregulated in clinical bladder cancer (BC) tumor tissues, The Cancer Genome Atlas (TCGA) database, and human BC cell lines. Inhibition of miR-3687 expression significantly reduced human BC cell proliferation in vitro and tumor growth in vivo, which coincided with the induction of G0/G1 cell cycle arrest and downregulation of cyclin E2 protein expression. Interestingly, overexpression of cyclin E2 reversed the inhibition of BC proliferation induced by miR-3687. Mechanistic studies suggested that miR-3687 binds to the 3' UTR of foxp1 mRNA, downregulates FOXP1 protein expression, and in turn promotes the transcription of cyclin E2, thereby promoting the growth of BC cells. Collectively, the current study not only establishes a novel regulatory axis of miR-3687/FOXP1 regarding regulation of cyclin E2 expression in BC cells, but also provides strong suggestive evidence that miR-3687 and FOXP1 may be promising targets in therapeutic strategies for human BC.

Detailed investigation of the iterative analysis for inertial confinement fusion target characterization.

The iterative algorithm based on optical path difference and ray deflection (IAORD) is investigated in detail, and an advanced version (AIAORD) is proposed to obtain the refractive indices of the shell and the ice layer of the inertial confinement fusion (ICF) target simultaneously. The concept of the fixed-point iteration is introduced in the advanced algorithm, and it is found that the right choice of the combination of the input values and the characteristic curves is the key to ensure convergence in the iteration. The test uncertainties of the index measurement are analyzed by simulations, and they show that the uncertainties of the refractive indices of the shell and ice layer are 9.94% and 1.20%, respectively. Characteristic curves of typical ICF targets are studied, from which we conclude that AIAORD is versatile and suitable for the applications with any two unknown target parameters to be solved.

Non-suture technique for rabbit oviduct anastomosis with 2-octyl cyanoacrylate: a histopathologic and biomechanical analysis.

AIM: The aim of this study was to investigate histological and biomechanical properties of oviduct anastomosis with 2-octyl cyanoacrylate (OCA) in the rabbit. MATERIAL AND METHODS: Sixty female rabbits were randomly divided equally into three groups: A (control), B (traditional catgut suture), and C (non-suture technique using OCA). After suture or OCA anastomosis, gross examination (adhesion formation) and histopathology (hematoxylin-eosin), ultrastructure (transmission electron microscopy), and biomechanics (bursting pressure) on para-anastomotic site were investigated on oviduct taken at 1 (A1, B1, C1) and 4 (A2, B2, C2) weeks, respectively. RESULTS: Adhesion score in group B was more severe than that in groups A and C at 1 and 4 weeks. Histopathology showed that acute endosalpingitis in group B was the most intense at 1 week, followed by significantly more tissue stimulation induced by catgut and foreign-body giant cells in group B than in group C at 4 weeks. Ultrastructural damage of ciliated cells was reversed partly (B2) and completely (C2) at 4 weeks. Bursting pressure in C1 was weaker than that in B1, followed by no significant difference at 4 weeks. CONCLUSION: Non-suture using OCA for oviduct anastomosis can be accepted as a new-perspective technique.

Inhibition of trauma-associated inflammatory liver damage by blocking NF-κB activity.

BACKGROUND/AIMS: NF-κB protein family members act as transcription facts and play a key role in regulating the immune response to infection and inflammatory signals. We proposed to determine the role of NF-κB in the development of trauma-associated liver damage and inflammation. METHODOLOGY: NF-κB DNA-binding activity was inhibited using double-stranded oligodeoxynucleotides (ODN). A total of 288 Wistar rats were randomly divided into four groups: control (C), traumatic inflammation (T), traumatic inflammation plus NF-κB decoy (ODN) and traumatic inflammation plus mutant NF-κB decoy ODN (mODN). RESULTS: Our data shows that inhibition of NF-κB activation significantly reduces liver tissue damage as evidenced by serum ALT levels and histological changes using both light microscopy and transmission electron microscopy. Furthermore, EMSA results showed that NF-κB activation was reduced in Group ODN rats compared to Group T and Group mODN rats. Expression of TNF-a and IL-6 protein in Group ODN rats were also reduced compared to Group T and Group mODN rats. We demonstrated that NF-κB plays an important role in trauma-associated inflammation and liver tissue damage. CONCLUSIONS: Suppressing NF-κB activation effectively reduces the release of the pro-inflammatory cytokines TNF-a and IL-6 following liver trauma.

[Expression of survivin, bcl-2 in cervical carcinoma and its association with HPV16/18 infection].

OBJECTIVE: To investigate the expression and the correlation of surviving, bcl-2 and HPV16/18 in cervical carcinoma. METHODS: Hybridization in situ was used to detect the expression of survivin mRNA and HPV16/18 DNA in 74 cases of CIN and 81 cases of cervical carcinoma and 20 cases of normal cervical tissues. And immunohistochemical analysis was used to detect the expression of bcl-2 protein. RESULTS: The positive rates of survivin mRNA, bcl-2 and HPV16/18 in CIN were 44.6%, 39.2% and 41.0% respectively versus 77.8%, 70.4% and 81.2% in cervical carcinoma. The above three indices gradually rose in normal cervical tissue, CIN and cervical carcinoma. The expression of survivin and bcl-2 in CINIII were obviously higher than those in CINI/II. And it was obviously higher in cervical carcinoma with stage IIb-III than those in stage I-IIa. And it was also obviously higher in cervical carcinoma with a poor differentiation than those with a good or medium differentiation. The expression of survivin in cervical carcinoma with lymphatic metastasis was significantly higher than that without lymphatic metastasis. There were no relationship between the expression of survivin or bcl-2 and the pathological type or tumor type of cervical carcinoma. The infection of HPV16/18 also had nothing to do with the clinical stage or pathological type or tumor type of cervical carcinoma. Inverse correlation was both observed in the expression of survivin and bcl-2 with survival rate. Thus a positive correlation between surviving, bcl-2 and HPV 16/18 was observed in cervical carcinoma. CONCLUSION: Survivin, bcl-2 and HPV16/18 participate in the development of cervical carcinoma. It may be a useful guide in early diagnosis of cervical carcinoma, evaluation of surgery and chemotherapy and prediction of outcome.

Prediction of overall survival time in patients with colon adenocarcinoma using DNA methylation profiling of long non-coding RNAs.

Long non-coding RNAs (lncRNAs) are a subgroup of RNAs able to regulate gene expression at the epigenetic level, and are therefore central to the regulation of numerous biological processes and the progression of multiple cancer types. However, lncRNAs have not been identified to considerably influence overall survival (OS) outcome in numerous different types of cancer. The majority of studies investigating the association between lncRNAs and epigenetic regulation have focused on their altered expression levels in cancerous cells, and few studies have focused on determining the correlation between lncRNAs and OS time. In the present study, comprehensive lncRNA expression analysis was performed on a cohort of patients diagnosed with colon adenocarcinoma (COAD) using the least absolute shrinkage and selection operator method (LASSO). Subsequently, the construction of a prognostic methylation-based predictive system was performed based on the results of LASSO analysis. Functional enrichment analysis of lncRNA co-expression genes was also performed. According to the results of the present study, the classifier was able to significantly predict the prognosis of patients with COAD, and the investigation of the relevant elucidated genes further revealed the mechanism of COAD pathogenesis.

Highly multiplexed quantifications of 299 somatic mutations in colorectal cancer patients by automated MALDI-TOF mass spectrometry.

BACKGROUND: Detection of somatic mutations in tumor tissues helps to understand tumor biology and guide treatment selection. Methods such as quantitative PCR can analyze a few mutations with high efficiency, while next generation sequencing (NGS) based methods can analyze hundreds to thousands of mutations. However, there is a lack of cost-effective method for quantitatively analyzing tens to a few hundred mutations of potential biological and clinical significance. METHODS: Through a comprehensive database and literature review we selected 299 mutations associated with colorectal cancer. We then designed a highly multiplexed assay panel (8-wells covering 299 mutations in 109 genes) based on an automated MADLI-TOF mass spectrometry (MS) platform. The multiplex panel was tested with a total of 319 freshly frozen tissues and 92 FFPE samples from 229 colorectal cancer patients, with 13 samples also analyzed by a targeted NGS method covering 532 genes. RESULTS: Multiplex somatic mutation panel based on MALDI-TOF MS detected and quantified at least one somatic mutation in 142 patients, with KRAS, TP53 and APC being the most frequently mutated genes. Extensive validation by both capillary sequencing and targeted NGS demonstrated high accuracy of the multiplex MS assay. Out of 35 mutations tested with plasmid constructs, sensitivities of 5 and 10% mutant allele frequency were achieved for 19 and 16 mutations, respectively. CONCLUSIONS: Automated MALDI-TOF MS offers an efficient and cost-effective platform for highly multiplexed quantitation of 299 somatic mutations, which may be useful in studying the biological and clinical significance of somatic mutations with large numbers of cancer tissues.

Clinical features and potential relevant factors of renal involvement in primary Sjögren's syndrome.

OBJECTIVE: To investigate distinct features of renal involvement in patients with primary Sjögren's syndrome (pSS) and to identify potential factors associated with renal involvement. METHODS: Four hundred and thrity-four pSS patients from the Rheumatology Department of the First Affiliated Hospital of Wenzhou Medical University from 2013 to 2017 were included in a cross-sectional study. Patients with renal involvement were compared with their age- and gender-matched controls (pSS without renal involvement). Demographic, clinical, histological, nephritic, immunological features of renal involvement in pSS were systematically analyzed. Possible factors related to renal involvement were identified using multivariate logistic regression analyses. RESULTS: One hundred and ninety-two pSS patients (88.48%) with renal involvement were women with mean age of nearly 58 years and mean disease duration of above 4 years. Clinical manifestation, serologic and immunological features and renal biopsy class of the pSS patients with renal involvement were presented. By multivariate analyses, xerophthalmia, histological positivity for lower salivary gland biopsy (LSGB), anti-SSA/Ro52-positive, reduced complement 3 (C3) levels, hypoalbuminemia and anemia retained significant association with renal involvement in pSS (all P < 0.05). CONCLUSION: In addition to LSGB pattern, anti-SSA/Ro52-positivity, reduced C3 levels, hypoalbuminemia and anemia, also indicate significant association with renal involvement in pSS. Therefore, early vigilance is required for patients with these clinical manifestations.

MicroRNA-3648 Is Upregulated to Suppress TCF21, Resulting in Promotion of Invasion and Metastasis of Human Bladder Cancer.

Although microRNAs (miRNAs) are well-known for their potential in cancer, the function and mechanisms of miR-3648 have barely been explored in any type of cancer. We show here that miR-3648 is upregulated in human BC tissues in comparison with adjacent non-tumor tissues. Functional studies showed that inhibition of miR-3648 expression in the human invasive BC UMUC3 and T24T cell lines decreased migration and invasion in vitro and suppressed lung metastasis in vivo, whereas miR-3648 overexpression promoted BC cell migration and invasion. A bioinformatics screen and mRNA 3' UTR luciferase reporter assay showed that transcription factor 21 (TCF21) was a direct target of miR-3648, and the results obtained from using a miR-3648 inhibitor revealed that miR-3648 inhibited TCF21 protein expression by reduction of its mRNA stability. Further, Kisspeptin 1 (KISS1) was identified as a TCF21 downstream effector responsible for miR-3648-mediated BC invasion and lung metastasis. Collectively, the present results suggest that miR-3648 is overexpressed and plays an oncogenic role in mediation of BC invasion and metastasis through directing the TCF21/KISS1 axis, revealing miR-3648 as a potential biomarker for BC prognosis and a target for BC therapy.

Coexpression of VEGF-C and COX-2 and its association with lymphangiogenesis in human breast cancer.

BACKGROUND: Lymphangiogenesis has become a new research frontier in tumor metastasis since the discovery of reliable lymphatic markers that have allowed observation and isolation of lymphatic endothelium. Cyclooxygenase-2 (COX-2) has been reported to be involved in the critical steps in carcinogenesis. However, possible role of COX-2 in lymphangiogenesis and lymphatic metastasis is still poorly understood. In present study, we aimed to investigate the relationship between vascular endothelial growth factor-C (VEGF-C) and COX-2 in human breast cancer, and correlations with lymphangiogenesis and prognosis. METHODS: Tissue samples of primary tumors from 70 patients undergoing intentionally curative surgical resections for breast cancer were immunohistochemically examined for VEGF-C, COX-2, and D2-40 expressions. The association between COX-2 and VEGF-C expressions and clinicopathological parameters as well as prognosis were analysised. To demonstrate the presence of proliferating lymphatic endothelial cells, 10 random cases with high LVD counts were selected for D2-40/Ki-67 double immunostaining. RESULTS: A significant correlation was found between the expression of VEGF-C and COX-2 (r = 0.529, P < 0.001), and both elevated VEGF-C expression and elevated COX-2 expression were associated with higher lymph vessel density (LVD), lymph node metastasis and D2-40 positive lymphatic invasion (LVI) as well as worse disease free survival (DFS) and overall survival (OS) in a univariate analysis. In the double immunostain for the lymph vessel marker D2-40 and the proliferation marker Ki-67, the results confirmed Ki-67-positive nuclei in a proportion of lymph vessel endothelial cells. CONCLUSION: There is indeed lymphangiogenesis in breast cancer, the most compelling evidence being the presence of proliferating lymphatic endothelial cells. VEGF-C and COX-2 are coexpressed and significantly associated with lymphangiogenesis and prognosis in invasive breast cancer. Suggesting COX-2 may up-regulate VEGF-C expression and thus promote lymph node metastasis via lymphangiogenesis pathway in human breast cancer.

Simple instruments facilitating achievement of transanal total mesorectal excision in male patients.

AIM: To assess the efficacy of a modified approach with transanal total mesorectal excision (taTME) using simple customized instruments in male patients with low rectal cancer. METHODS: A total of 115 male patients with low rectal cancer from December 2006 to August 2015 were retrospectively studied. All patients had a bulky tumor (tumor diameter ≥ 40 mm). Forty-one patients (group A) underwent a classical approach of transabdominal total mesorectal excision (TME) and transanal intersphincteric resection (ISR), and the other 74 patients (group B) underwent a modified approach with transabdominal TME, transanal ISR, and taTME. Some simple instruments including modified retractors and an anal dilator with a papilionaceous fixture were used to perform taTME. The operative time, quality of mesorectal excision, circumferential resection margin, local recurrence, and postoperative survival were evaluated. RESULTS: All 115 patients had successful sphincter preservation. The operative time in group B (240 min, range: 160-330 min) was significantly shorter than that in group A (280 min, range: 200-360 min; P = 0.000). Compared with group A, more complete distal mesorectum and total mesorectum were achieved in group B (100% vs 75.6%, P = 0.000; 90.5% vs 70.7%, P = 0.008, respectively). After 46.1 ± 25.6 mo follow-up, group B had a lower local recurrence rate and higher disease-free survival rate compared with group A, but these differences were not statistically significant (5.4% vs 14.6%, P = 0.093; 79.5% vs 65.1%, P = 0.130). CONCLUSION: Retrograde taTME with simple customized instruments can achieve high-quality TME, and it might be an effective and economical alternative for male patients with bulky tumors.

Overexpression of salusin-ß is associated with poor prognosis in ovarian cancer.

Ovarian cancer is recognized as one of the worst gynecologic malignancies associated with rapid metastasis and poor overall survival rate. The identified valuable molecular biomarkers criticize importance of timely diagnosis for ovarian cancer. Salusin-ß levels are dramatically increased in women with polycystic ovarian syndrome. However, the roles of salusin-ß in ovarian cancer have yet to be fully elucidated. A total of 57 paired ovarian cancer specimens and matched adjacent normal tissues were used to measure the salusin-ß levels. The prognostic value of salusin-ß for tumor progression and survival rate was investigated. The effects of salusin-ß on ovarian cancer cell proliferation and epithelial-mesenchymal transition were also explored. The expression of salusin-ß was significantly increased in ovarian cancer tissue specimens compared with matched normal adjacent tissue (P<0.05). The high salusin-ß level was closely related with FIGO stage and lymph node metastases. The ovarian cancer patients with high salusin-ß had a shorter overall survival (P<0.05). Salusin-ß obviously enhanced the proliferation and epithelial mesenchymal-transition of SKOV3 cells. Furthermore, salusin-ß substantially decreased the expression of p-GSK-3ß and GSK-3ß, but stimulated the ß-catenin expression and downstream genes of wnt/ß-catenin including cyclin D1 and C-myc. Our data demonstrated for the first time that upregulated salusin-ß may be a novel independent prognostic biomarker for overall survival of ovarian cancer. Salusin-ß accelerated the proliferation and epithelial mesenchymal transition of ovarian cancer cells at least partly via activation of Wnt/ß-catenin signaling pathway. Salusin-ß may be an important target for therapeutic intervention in ovarian cancer.

Impact of platelet to lymphocyte ratio and metabolic syndrome on the prognosis of colorectal cancer patients.

OBJECTIVE: The aim of this study was to evaluate the prognostic value of both platelet to lymphocyte ratio (PLR) and metabolic syndrome (MetS) in colorectal cancer (CRC) patients. PATIENTS AND METHODS: We retrospectively enrolled 1,163 CRC patients. Preoperative values of PLR were stratified into three groups according to cut-off values of 120 and 220. The Kaplan-Meier analysis was used to calculate cumulative survival rate related to PLR and MetS. Cox proportional hazard regression models were used to analyze potential risk factors and the prognosis associated with PLR and MetS in CRC patients. RESULTS: PLR was significantly higher in the MetS(+) group as compared to MetS(-) group (P=0.039). An elevated PLR was significantly associated with mortality (P=0.014), but not the existence of MetS (P=0.235). In multivariate regression analysis, PLR was an independent risk factor for overall survival (OS) (P=0.046). For the subgroup with a PLR >220, MetS was an independent predictor for both OS and disease-free survival (P=0.039 and P=0.047, respectively) by multivariate analysis adjusting for confounding covariates. In addition, the presence of MetS was associated with a 2-fold increased risk of mortality and tumor recurrences (hazard ratio [HR] =2.0 and HR =1.9, P<0.05, respectively). CONCLUSION: Preoperative PLR was associated with MetS in CRC patients. Testing for the combined presence of PLR and MetS could potentially improve the predictive accuracy of CRC prognosis.

ß3-tubulin is a good predictor of sensitivity to taxane-based neoadjuvant chemotherapy in primary breast cancer.

The objective of this study was to explore the relationship between ß3-tubulin expression and sensitivity to taxane-based neoadjuvant chemotherapy in primary breast cancer patients. A total of 48 local advanced breast cancer patients that received taxane-containing neoadjuvant chemotherapy were studied. The levels of ß3-tubulin expression were tested by immunohistochemistry before chemotherapy and at the end of cycles 2, 4 and 6. The correlation between the efficacy of the chemotherapy and ß3-tubulin expression and changes in ß3-tubulin expression over the course of chemotherapy was examined. ß3-tubulin protein expression before chemotherapy was significantly and negatively correlated with the response rate. The overall response rate was 31.8 % in the high ß3-tubulin expression group, whereas it was 84.6 % in the low ß3-tubulin expression group. At the end of cycles 2, 4 and 6 during the treatment course, the average expression rates of ß3-tubulin were showed an increasing trend with ß3-tubulin expression level at the end of cycle 4 being significantly different from that before chemotherapy. Nine patients that had a low ß3-tubulin expression level preneoadjuvant chemotherapy changed to a high ß3-tubulin expression level postneoadjuvant chemotherapy, and they had lower response rate than patients with consistent low. In conclusion, ß3-tubulin is a good predictor of chemosensitivity to taxane for breast cancer, and the change of its expression level during chemotherapy may be an important cause of secondary resistance to taxane. Detection of ß3-tubulin expression before and throughout the chemotherapy will help with selection of the chemotherapy treatment plan.

Pathologically decreased expression of miR-193a contributes to metastasis by targeting WT1-E-cadherin axis in non-small cell lung cancers.

BACKGROUND: The metastatic cascade is a complex and multistep process with many potential barriers. Recently, miR-193a has been reported to be a suppressive miRNA in multiple types of cancers, but its underlying anti-oncogenic activity in non-small cell lung cancers (NSCLC) is not fully elucidated. METHODS: The expressions of miR-193a (miR-193a-5p) in human lung cancer tissues and cell lines were detected by real-time PCR. Dual-luciferase reporter assay was used to identify the direct target of miR-193a. Cell proliferation, apoptosis, and metastasis were assessed by CCK-8, flow cytometry, and Transwell assay, respectively. RESULTS: The expression of miR-193a in lung cancer tissues was decreased comparing to adjacent non-tumor tissues due to DNA hypermethylation in lung cancer tissues. Ectopic expression of miR-193a inhibited cell proliferation, colony formation, migration, and invasion in A549 and H1299 cells. Moreover, overexpression of miR-193a partially reversed tumor growth factor-ß1 (TGF-ß1)-induced epithelial-to-mesenchymal transition (EMT) in NSCLC cells. Mechanistically, miR-193a reduced the expression of WT1, which negatively regulated the protein level of E-cadherin, suggesting that miR-193a might prevent EMT via modulating WT1-E-cadherin axis. Importantly, knockdown of WT1 resembled the anti-cancer activity by miR-193a and overexpression of WT1 partially reversed miR-193a-induced anti-cancer activity, indicating that WT1 plays an important role in miR-193a-induced anti-cancer activity. Finally, overexpression of miR-193a decreased the growth of tumor xenografts in mice. CONCLUSION: Collectively, our results have revealed an important role of miR-193a-WT1-E-cadherin axis in metastasis, demonstrated an important molecular cue for EMT, and suggested a therapeutic strategy of restoring miR-193a expression in NSCLC.

Tumor M2 pyruvate kinase in diagnosis of nonsmall cell lung cancer: A meta-analysis based on Chinese population.

OBJECTIVE: The purpose of this study was to evaluate the value of tumor M2 pyruvate kinase (tumor M2-PK) in the diagnosis of nonsmall cell lung cancer. METHODS: The diagnosis clinical studies of tumor M2-PK in the diagnosis of nonsmall cell lung cancer were electronic researched in the Medline, EMBASE, WANFANG, and CNIK databases. The data of true positive, false positive, false negative, and true negative were extracted from each of the individual studies. We use  Stata11.0 (http://www.stata.com; Stata Corporation, College Station, TX) and MetaDiSc 1.4 software to pool the diagnostic sensitivity, specificity, and diagnostic area under the receiver operating characteristic (ROC). RESULTS: Eleven diagnostic clinical studies with 1294 subjects were included in this diagnostic meta-analysis. The combined sensitivity, specificity, positive likely hood ratio, negative likely hood ratio were 0.69 (0.65-0.72), 0.92 (0.89-0.94), 7.84 (5.92-10.38), 0.36 (0.32-0.40). And the area under the ROC curve was 0.92 (0.90-0.94). CONCLUSION: Serum tumor M2-PK can be a potential biomarker for diagnosis of nonsmall cell lung cancer.

mTOR activation in immature cells of primary nasopharyngeal carcinoma and anti-tumor effect of rapamycin in vitro and in vivo.

The mammalian target of rapamycin (mTOR) signaling is a key pathway in the progression of different cancers and in the homeostasis of stem cells. Here, we investigated the link between mTOR signaling and cancer stem cells (CSCs) in nasopharyngeal carcinoma (NPC). We found that human primary NPC expressed embryonic stem cell (ESC) markers: CD133, SOX2 and OCT4 as well as pmTOR and pS6. Primary ESC-positive NPC cells could form secondary NPC in BALB/c nude mice. Rapamycin, an mTOR inhibitor, significantly suppressed ESC-positive NPC cell growth in vitro and tumor formation in vivo. Our findings suggest that mTOR signaling is activated in CSC-like cells and plays an important role in NPC growth.