Li Ren

Background: Noninvasive biomarkers for the assessment of response to chemotherapy in advanced breast cancer (BCa) are essential for optimized therapeutic decision-making. We evaluated the potential of soluble Periostin (POSTN) in circulation as a novel biomarker for chemotherapy efficacy monitoring. Methods: Two hundred and thirty-one patients with different stages of BCa were included. Of those patients, 58 patients with inoperable metastatic disease receiving HER2-targeted or non-targeted chemotherapy were enrolled to assess the performances of markers in recapitulating the chemotherapy efficacy assessed by imaging. POSTN, together with CA153 or CEA at different time points (C0, C2, and C4) were determined. Results: POSTN levels were significantly associated with tumor volume (P < 0.0001) and TNM stages (P < 0.0001) of BCa. For early monitoring, dynamics of POSTN could recapitulate the chemotherapy efficacy among all molecular subtypes (Cohen’s weighted kappa = 0.638, P < 0.0001), much better than that of carcinoembryonic antigen (CEA) and cancer antigen 153 (CA15-3). For early partial response, superior performance of POSTN was observed (Cohen’s weighted kappa = 0.827, P < 0.0001) in cases with baseline levels above 17.19 ng/mL. For long-term monitoring, the POSTN response was observed to be strongly consistent with the course of the disease. Moreover, progression free survival analysis showed that patients experienced a significant early decrease of POSTN tended to obtain more benefits from the treatments. Conclusions: The current study suggests that soluble POSTN is an informative serum biomarker to complement the current clinical approaches for early and long-term chemotherapy efficacy monitoring in advanced BCa.

Tumour cells mainly generate energy from glycolysis, which is commonly coupled with lactate production even under normoxic conditions. As a critical lactate transporter, monocarboxylate transporter 4 (MCT4) is highly expressed in glycolytic tissues, such as muscles and tumours. Overexpression of MCT4 is associated with poor prognosis for patients with various tumours. However, how MCT4 function is post-translationally regulated remains largely unknown. Taking advantage of human lung adenocarcinoma (LUAD) cells, this study revealed that MCT4 can be polyubiquitylated in a nonproteolytic manner by SYVN1 E3 ubiquitin ligase. The polyubiquitylation facilitates the localization of MCT4 into the plasma membrane, which improves lactate export by MCT4; in accordance, metabolism characterized by reduced glycolysis and lactate production is effectively reprogrammed by SYVN1 knockdown, which can be reversed by MCT4 overexpression. Biologically, SYVN1 knockdown successfully compromises cell proliferation and tumour xenograft growth in mouse models that can be partially rescued by overexpression of MCT4. Clinicopathologically, overexpression of SYVN1 is associated with poor prognosis in patients with LUAD, highlighting the importance of the SYVN1-MCT4 axis, which performs metabolic reprogramming during the progression of LUAD.

Tumor cells mainly generate energy from glycolysis that is commonly coupled with lactate production even under normoxic conditions. As one critical lactate transporter, monocarboxylate transporter 4 (MCT4) is highly expressed in glycolytic tissues like muscles and tumors. Overexpression of MCT4 is associated with poor prognosis for patients with various tumors. However, it remains largely unknown how MCT4 function is post-translationally regulated. Taking advantage of human lung adenocarcinoma (LUAD) cells, this study reveals MCT4 can be polyubiquitylated in a non-proteolytic manner by SYVN1 E3 ubiquitin ligase. The polyubiquitylation facilitates the localization of MCT4 into the plasma membrane that enhances the activity of MCT4 to export lactate; in accordance, SYVN1 knockdown effectively reprograms metabolism that is characterized by reduced glycolysis and lactate production, which can be reversed by ectopic MCT4 expression. Biologically, SYVN1 knockdown successfully compromises cell proliferation and tumor xenograft growth in mouse models. Clinicopathologically, overexpression of SYVN1 is associated with poor prognosis in patients with LUAD, highlighting the importance of SYVN1-MCT4 axis contributes to the progression of LUAD through metabolic reprograming.

Background: Breast cancer (BCa) is the leading cause of cancer deaths among women. Reliable biomarkers for early diagnosis and metastasis prediction are essential to improve the prognosis of BCa. This study aimed to evaluate serum periostin (POSTN) as a novel biomarker complementing CA153 (carbohydrate antigen 153) and CEA (carcinoembryonic antigen) for BCa diagnosis and metastasis prediction. Methods: To assess the potential of soluble POSTN as a circulating biomarker, 242 participants, including 173 patients with different stages of BCa and 69 healthy individuals, were enrolled in this study. Soluble POSTN, together with CA153 and CEA, were determined in serum by enzyme linked immunosorbent assay (ELISA) or electrochemiluminescence immunoassays. Results: Serum POSTN levels in locoregional BCa patients were significantly higher than that in healthy controls. Receiver operating curve (ROC) analysis revealed that, to distinguish health controls from locoregional BCa, POSTN was observed with the highest AUC (area under curve) (AUC_POSTN = 0.72 [0.65 – 0.79], AUC_CA153 = 0.57 [0.49 – 0.64], AUC_CEA = 0.62 [0.55 – 0.69]), and both CA153 and CEA were observed with significantly improved AUCs by combination with POSTN (AUC_{POSTN + CA153} = 0.74 [0.67 – 0.80], P < 0.001; AUC_{POSTN + CEA} = 0.77 [0.70 – 0.82], P < 0.001). Moreover, the performances of the POSTN were comparable with that of CA153 in predicting distant metastasis of BCa (AUC_POSTN = 0.78 [0.71 – 0.84], AUC_CA153 = 0.82 [0.76 – 0.88]). Kaplan–Meier analysis indicated that elevated serum POSTN was associated with poor overall survival and progression-free survival. Conclusions: This study suggested that soluble POSTN is a promising potential biomarker for diagnosis and metastasis prediction of BCa.

Hepatocellular carcinoma (HCC) is highly malignant; nearly half of the new cases and deaths are in China. The poor prognosis of HCC is mainly due to late diagnosis; many new biomarkers have been developed for HCC diagnosis. However, few markers are quickly translated into clinical practice; early and differential diagnosis of HCC from cirrhosis and/or hepatitis is still a clinical challenge. Metabolomics and biochemical methods were used to reveal specific serum biomarkers of HCC. Most of the elevated metabolites in HCC and HBV patients were overlapped compared with controls. Urea was the specifically elevated serum biomarker of HCC patients. Moreover, urea combined with AFP and CEA can improve the sensitivity of HCC diagnosis. The plasma ammonia of HCC patients was significantly higher than healthy controls. Co-culture cell model revealed normal liver cells cooperated with cancer cells to metabolize ammonia into urea. The urea metabolism in cancer cells marginally depended on the expression of CPS1. However, the expression of CPS1 did not change with ammonium chloride, which might regulate the urea cycle through enzyme activity. The urea cycle could detoxify high concentrations of ammonia to promote cancer cell proliferation. Therefore, urea was a by-product of ammonia metabolism and could be a potential serum biomarker for HCC. The combined application of metabolomics and biochemical methods can discover new biomarkers for the early diagnosis of HCC and be quickly applied to clinical diagnosis.

Background: Although hematologic review criteria for general hospitals have been established, they may be insufficient for cancer hospitals. This study aimed to establish the appropriate review criteria for hematology analyzers in cancer hospitals. Methods: A total of 1003 samples from our hospital were randomly selected for blood smear preparation and microscopic review. The review criteria of the International Consensus Group for Hematology Review (ICGH) and Chinese consensus group were used to obtain the review, true‐negative (TN), true‐positive (TP), false‐negative (FN), and false‐positive (FP) rates, as well as the triggered rules. Our review criteria were established by comparing flag or numeric value information of TP and FP samples, adjusting rules to obtain better efficiency, a low slide review rate, and an acceptable FN rate. Results: Overall, 197 (19.64%) samples showed positive smear findings. Compared to the ICGH criteria, the slide review rate of the newly established criteria declined from 51.25% to 39.28%, and the TP and TN rates increased from 17.85% and 46.06% to 23.13% and 55.83%, respectively. The FN rate of the newly established criteria was 3.69%. Another set of samples used to validate the newly established criteria yielded the review, FN, and FP rates as 33.49%, 1.86%, and 25.58%, respectively. Conclusion: The newly established review criteria for hematology analyzers enabled the prompt identification, smear, and further verification of doubtful specimens, without a significant increase in the workload, thus improving the efficiency of the review process. This study provided data support for other cancer hospitals to establish review criteria.

Pancreatic cancer has a low survival rate, and most patients have lymph node metastasis and distant metastasis at the time of diagnosis. Despite efforts to improve overall survival (OS) and recurrence free survival (RFS), the prognosis of pancreatic cancer remains poor, underscoring the importance of identifying new biomarkers to predict metastasis in patients with pancreatic cancer. Leukemia inhibitory factor (LIF) is overexpressed in many types of cancer and is involved in the development of various malignancies including pancreatic cancer. However, the role of LIF as a biomarker to predict metastasis in pancreatic cancer remains unclear. In this study, univariate and multivariate Cox regression analyses identified LIF expression in pancreatic tumor tissues as an independent risk factor related to worse OS and RFS. LIF overexpression was related to poor clinicopathological features such as lymph node metastasis and Pathological stage (pTNM) stage. Serum LIF levels were higher in pancreatic cancer patients than in healthy controls. The area under the receiver operating characteristic curve indicated that serum LIF is more effective than other biomarkers (CA199 and CEA) for predicting lymph node and distant metastasis.

Background: Hypoxia is a hallmark of cancer and is associated with poor prognosis. However, the molecular mechanism by which hypoxia promotes tumor progression remains unclear. MicroRNAs dysregulation has been shown to play a critical role in the tumor and tumor microenvironment. Here, we investigated the roles of miR‐495 and miR‐5688 in human non‐small cell lung cancer (NSCLC) and their underlying mechanism. Methods: The expression levels of miR‐495 and miR‐5688 in human NSCLC tissue specimens were measured by quantitative real‐time polymerase chain reaction (qRT‐PCR). Deferoxamine (DFO) was used to determine whether the regulation of miR‐495 and miR‐5688 under hypoxia was dependent on hypoxia‐inducible factor 1‐alpha (HIF‐1α). Furthermore, the functions of miR‐495 and miR‐5688 in tumor progression were evaluated using colony formation, 3‐(4,5‐dimethylthiazol‐2‐yl)‐5‐(3‐carboxymethoxyphenyl)‐2‐(4‐sulfophenyl)‐2H‐tetrazolium (MTS), wound healing, transwell assays, and xenograft model. Two algorithms, PicTAR and Targetscan, were used to predict the target gene of these two miRNAs, and dual‐luciferase reporter assay was conducted to confirm the target. The unpaired two‐tailed t test, Pearson correlation analysis, and Fisher's exact probability test were performed for statistical analyses. Results: Two miRNAs, miR‐495 and miR‐5688, were found to participate in NSCLC progression under hypoxia. They were down‐regulated in NSCLC tissues compared with normal tissues. We determined that hypoxia led to the down‐regulation of miR‐495 and miR‐5688 in NSCLC cells, which was independent of HIF‐1α and cellular metabolic energy. In addition, miR‐495 and miR‐5688 suppressed cell proliferation, migration, and invasion in vitro . The NSCLC xenograft model showed that miR‐495 and miR‐5688 inhibited tumor formation in vivo . Interestingly, we found that miR‐495 and miR‐5688 had the same target, interleukin‐11 (IL‐11). Recombinant human IL‐11 counteracted the effects of miR‐495 and miR‐5688 on NSCLC cells, suggesting that miR‐495 and miR‐5688 executed their tumor suppressive role by repressing IL‐11 expression. Conclusion: We found that hypoxia down‐regulated the expression levels of miR‐495 and miR‐5688 in NSCLC to enhance IL‐11 expression and tumor progression, indicating that the miR‐495/miR‐5688/IL‐11 axis may serve as a therapeutic target and potential biomarker for NSCLC.

Objective Mitochondria play multifunctional roles in carcinogenesis. Deciphering uncertainties of molecular interactions within mitochondria will promote further understanding of cancer. Interleukin enhancer binding factor 2 (ILF2) is upregulated in several malignancies, however, much remains unknown regarding ILF2 in small cell lung cancer (SCLC). In the current study, we explored ILF2’s role in SCLC and demonstrated its importance in mitochondria quality control. Methods Colony formation, cell proliferation, cell viability and xenograft studies were performed to examine ILF2’s role on SCLC progression. Glucose uptake, lactate production, cellular oxygen consumption rate and extracellular acidification rate were measured to examine the effect of ILF2 on glucose metabolism. RNA-sequencing was utilized to explore genes regulated by ILF2. E2F1 transcriptional activity was determined by dual luciferase reporter assay. Mitochondria quantification and mitochondrial membrane potential assays were performed to examine mitochondrial quality. Gene expression was determined by RT-qPCR, Western blot and IHC assay. Results ILF2 promotes SCLC tumor growth in vitro and in vivo. ILF2 elevates oxidative phosphorylation expression and declines glucose intake and lactate production. Genome-wide analysis of ILF2 targets identified a cohort of genes regulated by E2F1. In consistent with this, we found ILF2 interacts with E2F1 in SCLC cells. Further studies demonstrated that suppression of E2F1 expression could reverse ILF2-induced tumor growth and enhanced mitochondria function. Significantly, expression of ILF2 is progressively increased during SCLC progression and high ILF2 expression is correlated with higher histologic grades, which indicates ILF2’s oncogenic role in SCLC. Conclusions Our results demonstrate that ILF2 interacts with E2F1 to maintain mitochondria quality and confers SCLC cells growth advantage in tumorigenesis.

Background: Hypoxia is present in many solid tumors and is a prognostic factor for poor tumorigenesis. Here, we investigated whether carcinoembryonic antigen (CEA) plays a role in the hypoxic microenvironment of breast cancer. Methods: We researched the expression of CEA in breast tumor tissues and the corresponding paracancerous tissues. Furthermore, western blotting was used to detect the expression levels of CEA in breast cancer cells cultured under hypoxia or normoxia. The expression levels of CEA, hypoxia-inducible factor-1 alpha (HIF-1 alpha) and carbonic anhydrase IX (CA-IX) in tumor tissues were detected by immunohistochemical staining. In addition, the correlations between CEA and clinical parameters and prognosis of breast cancer patients were analyzed by statistical analysis. Results: We found that CEA mRNA expression of tissues in breast cancer patients was significantly increased compared to the corresponding paracancerous tissues. Furthermore, the expression levels of CEA in breast cancer cells cultured under hypoxic conditions were elevated compared with those cultured under normoxic conditions. Immunohistochemical staining demonstrated that the expression of CEA in tumor tissues was increased with HIF-1 alpha, and the area of CA-IX expression could be seen with CEA distribution. In addition, the clinical parameter analysis found that serum CEA was significantly associated with malignant clinical features. We also found that elevated CEA expression was related to poor prognosis in breast cancer patients. Conclusions: CEA expression was elevated in the breast cancer hypoxic microenvironment, and the data suggested a rationale for the use of CEA in breast cancer diagnosis, assessment, and prognosis.